Publications by authors named "Zegeng Li"

28 Publications

  • Page 1 of 1

[Rapid identification of chemical components in Qi-Yu-San-Long decoction by ultra high performance liquid chromatography-quadrupole time-of-flight mass spectrometry].

Se Pu 2021 Jul;39(7):730-743

Scientific Research & Experiment Center, Anhui University of Chinese Medicine, Hefei 230038, Chin.

Qi-Yu-San-Long decoction (QYSLD) is a classic traditional Chinese medicine prescription consisting of ten types of herbal medicines, including Astragali Radix, Polygonati Odorati Rhizoma, Scolopendra, Pheretima, L., Willd., Coicis Semen, L., Curcumae Rhizoma, and Fritillariae Cirrhosae Bulbus, combined in a ratio of 15∶5∶3∶3∶10∶10∶10∶3∶5∶3 by weight. QYSLD has been used to treat non-small cell lung cancer (NSCLC) for over 20 years in clinical practice, and its curative effect is considered credible. However, the chemical constituents of QYSLD have not been revealed because of their complexity, which has significantly hindered the systematic clarification of the efficacy of the materials and quality evaluation. In this study, a reliable strategy based on the data-independent acquisition (DIA) technology of ultra high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) combined with a targeted screening method was established to investigate the chemical components of QYSLD. A 2-μL aliquot from each vial was injected into a Waters ACQUITY UPLC BEH C column (100 mm×2.1 mm, 1.7 μm) to separate complex components. The temperature of the column was 35 ℃, and the flow rate was set at 0.2 mL/min. The mobile phase consisted of 0.1% formic acid aqueous solution and acetonitrile. Detection was conducted using an Xevo G2-XS QTOF-MS with a LockSpray capable-electrospray interface. The data for complex components in QYSLD were collected by full-information tandem mass spectrometry (MS ) in the positive and negative ion modes. In the MS mode, data acquisition was performed using a mass spectrometer by rapidly switching from a low-collision-energy (CE) scan to a high-CE scan during a single LC run. Thus, accurate precursor and fragment ions were collected in a single run, which was helpful for the structural elucidation of multiple components in QYSLD. In addition, systematic information on isolated chemical compounds was collected and distinguished from the ten individual herbs in QYSLD using databases such as China Academic Journals Full-text database (CNKI), PubMed, Web of Science, Medline, and ChemSpider. Accordingly, a self-building library of QYSLD, including the component name, molecular formula, and structure of the components from the herbs, was established. Subsequently, the raw MS data of the collected samples and the self-building chemical composition library were imported into a natural product post-processing screening (UNIFI) platform for targeted screening of the chemical components in QYSLD. The parameters for UNIFI platform were as follows: the retention time deviation was ±0.1 min; an error margin of no more than 5×10 for the identified compounds was allowed; positive adducts, including [M+H]and [M+Na], were selected; and negative adducts, including [M-H] and [M+HCOO], were selected. The results showed that a total of 166 compounds were initially identified, including 22 saponins, 13 alkaloids, 27 flavonoids, 32 terpenes, 20 amino acids, 16 phenylpropanoids, 9 organic acids, 6 sterols, 6 anthraquinones, and 15 other components. Among them, sixteen components were confirmed unambiguously with the reference substances. To better understand the chemical contribution of individual herbs to the entire decoction, the attributes of each component were summarized. This study provides a foundation for exploring the pharmacodynamic substances of QYSLD.
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http://dx.doi.org/10.3724/SP.J.1123.2020.10016DOI Listing
July 2021

Qiyusanlong Formula Induces Autophagy in Non-Small-Cell Lung Cancer Cells and Xenografts through the mTOR Signaling Pathway.

Evid Based Complement Alternat Med 2021 22;2021:5575453. Epub 2021 May 22.

Institute of Traditional Chinese Medicine of Respiratory Disease Prevention, Anhui University of Chinese Medicine, Hefei, Anhui, China.

Objective: Qiyusanlong (QYSL) formula has been used in the clinic for more than 20 years and has been proved to have pronounced efficacy in the treatment of non-small-cell lung cancer (NSCLC). This work aims to evaluate the molecular mechanism of QYSL formula action on NSCLC, specifically in relation to autophagy induction.

Methods: In vitro, CCK-8 was used to detect the effect of QYSL serum on cell viability in A549 cells. In vivo, A549 cells were implanted subcutaneously in nude mice to establish a xenograft model. TUNEL staining was used to measure cell apoptosis and TEM to observe the autophagy-related morphological changes in vitro and in vivo. Western blotting, RT-qPCR, and immunofluorescence were used to measure autophagy-related proteins. In addition, rapamycin (an inhibitor of mTOR and inducer of autophagy) and MHY1485 (an activator of mTOR and inhibitor of autophagy) were used to determine whether QYSL-induced autophagy was regulated by the mTOR pathway.

Results: QYSL serum inhibited the cell viability of A549 cells in a concentration-dependent manner. In vivo, the QYSL formula inhibited xenograft growth. The QYSL formula promoted apoptosis in A549 cells and induced autophagosome formation in vitro and in vivo. In addition, the QYSL formula downregulated the expression of mTOR and p62, while it upregulated the expression of ATG-7 and Beclin-1 and increased the LC3-II/LC3-I ratio. QYSL serum inhibited p-mTOR in a similar manner to rapamycin while reducing the activating effects of MHY1485 on p-mTOR.

Conclusion: The QYSL formula has anti-lung cancer effects and promotes autophagy through the mTOR signaling pathway.
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http://dx.doi.org/10.1155/2021/5575453DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8164545PMC
May 2021

Allyl isothiocyanate increases MRP1 expression in cigarette smoke extract-stimulated human bronchial epithelial cells via the JNK/Nrf2 pathway.

Exp Ther Med 2021 Apr 25;21(4):409. Epub 2021 Feb 25.

School of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui 230012, P.R. China.

Multidrug resistance-related protein 1 (MRP1) is involved in the biological transport of several molecules with diverse structural characteristics outside of the cell. In addition to its transport activity, MRP1 exhibits multiple defense mechanisms . MRP1 is highly expressed in normal lung tissues and plays a protective role in the process of chronic obstructive pulmonary disease. In the present study, human bronchial epithelial cells (16HBE14o-cells) were stimulated by cigarette smoke extract (CSE) to simulate a smoking environment. On this basis, the mechanism of Allyl isothiocyanate (AITC) administration on the expression of MRP1 in CSE-stimulated 16HBE14o-cells was investigated. The effects of CSE on the viability of 16 HBE14o-cells were investigated by an MTT assay. The changes in the mRNA expression levels of nuclear erythroid factor 2 (Nrf2) and MRP1 were investigated in CSE-stimulated 16HBE14o-cells using western blotting and reverse transcription quantitative PCR (RT-qPCR). Immunofluorescence analysis was used to detect Nrf2 nuclear translocation. Incubation of the cells with 5% CSE for 24 h had minor effects on cell viability and resulted in the activation of the JNK and p38MAPK signaling pathways. AITC activated the JNK pathway, inhibited the activation of the p38MAPK pathway in 16HBE14o-cells stimulated by 5% CSE and upregulated the expression levels of Nrf2 and MRP1 in a time-dependent manner. The upregulation of Nrf2, MRP1 and of Nrf2, and MRP1 mRNA expression levels in CSE-stimulated cells was inhibited by pretreatment with SP600125 (a JNK pathway inhibitor). Furthermore, the fluorescence intensity in the nucleus was significantly enhanced following AITC pretreatment and the analysis indicated nuclear translocation of Nrf2 in the cells. These results indicated that Nrf2 and MRP1 expression levels in CSE-stimulated cells were altered following AITC pretreatment. Thus demonstrating that the primary mechanism may be associated with activation of the JNK pathway, while the p38MAPK pathway may not be involved.
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http://dx.doi.org/10.3892/etm.2021.9840DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7938453PMC
April 2021

Identification and Characterization of Chemical Constituents in HuaTanJiangQi Capsules by UPLC-QTOF-MS Method.

J AOAC Int 2021 Aug;104(4):983-998

Department of Respiratory Medicine, The First Affiliated Hospital to Anhui University of Chinese Medicine, Hefei, 230012, Anhui, China.

Background: HuaTanJiangQi (HTJQ) is a classical Chinese medicine compound preparation, mainly used for clinically treating and improving chronic obstructive pulmonary disease (COPD) in China.

Objective: To establish a rapid and efficient analytical method for the identification and characterization of chemical constituents in HTJQ based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS).

Method: UPLC-QTOF-MS was used to rapidly separate and identify the chemical constituents of HTJQ via a gradient elution system. The accurate mass data of the protonated and deprotonated molecules and fragment ions were detected in positive and negative ion modes. Compounds of HTJQ can be identified and assigned by analyzing accurate mass measurements and ion fragmentation mechanisms and comparing them with a chemical compositions database.

Results: A total of 61 compounds in HTJQ were separated and identified, including 14 flavonoids, 16 organic acids, four isothiocyanic acids, eight butyl phthalides, two alkaloids, 10 terpenoids, four methoxyphenols and furanocoumarins, and three other compounds. The chemical compounds of HTJQ were identified and elucidated comprehensively for the first time.

Conclusions: A rapid, accurate, and efficient UPLC-QTOF-MS method has been developed for the identification of chemical components and applied to simultaneously evaluate the quality and effectiveness of HTJQ.

Highlights: Systematic identification of chemical constituents in HTJQ can provide a scientific and reasonable basis for the application of HTJQ in the clinical treatment of COPD.
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http://dx.doi.org/10.1093/jaoacint/qsab004DOI Listing
August 2021

[Bioinformatical analysis of differentially expressed genes in alveolar macrophages of chronic obstructive pulmonary disease(COPD)].

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2020 Nov;36(11):961-966

Institute of Medicine for Respiratory Diseases, Anhui Academy of Chinese Medicine, Department of Respiratory Diseases, First Affiliated Hospital, Anhui University of Chinese Medicine, Hefei 230031, China. *Corresponding author, E-mail:

Objective To analyze the differentially expressed genes (DEGs) in alveolar macrophages (AMs) of patients with chronic obstructive pulmonary disease (COPD) and their potential roles in the pathogeneses of COPD using bioinformatics. Methods Gene chip and RNA sequencing data sets of AMs in patients with COPD were downloaded from GEO. Limma and Degseq2 packages in R software were applied to obtain DEGs, and the GO enrichment analysis, the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, protein-protein interaction analysis (PPI), and the hub gene analysis were performed to predict the molecular mechanisms of DEGs. Results Through the integration of three data sets, a total of 43 DEGs of AMs were obtained, and the function predictive analysis found that the 43 DEGs were primarily related to chemokines, cytokines, complement, cytochrome P450, etc., which mainly included the significantly low expression of C-X-C motif chemokine ligand 9 (CXCL9), CXCL11, etc. and the significantly high expression of cytochrome P450 family 1 subfamily B member 1 (CYP1B1). Conclusion The DEGs of AMs in patients with COPD are related to the molecular mechanisms of immunity and inflammation and might be involved in the pathogenesis of chronic inflammation of COPD.
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November 2020

Paeonol loaded cyclodextrin metal-organic framework particles for treatment of acute lung injury via inhalation.

Int J Pharm 2020 Sep 13;587:119649. Epub 2020 Jul 13.

Center for Drug Delivery System, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, China; Anhui University of Chinese Medicine, Hefei 230012, China; NMPA Key Laboratory for Quality Research and Evaluation of Pharmaceutical Excipients, National Institutes for Food and Drug Control, No. 2 Tiantan Xili, Beijing 100050, China. Electronic address:

Administration of drugs via inhalation can overcome issues related to poor water solubility, low absorption or bioavailability associated with oral administration. In the current study we used γ-cyclodextrin metal organic frameworks (CD-MOFs) - of inhalable particle sizes, with cubic morphologies and favorable aerodynamic properties to achieve targeted pulmonary drug delivery via dry powder inhalers. The active natural compound, paeonol (PAE), was chosen as a model drug for treatment of acute lung injury (ALI). After loading of PAE into CD-MOF particles of inhalable sizes, PAE was rapidly released in phosphate buffer (pH = 7.4; 90% within 30 min) and in vivo experiments. After mixing with coarse and fine lactose (Inhalac®230: Inhalac®400: PAE-CD-MOF = 40:10:50, w/w/w), paeonol had a high fine particle fraction (FPF) (28.59%). Atom force microscopy was used to assess surface roughness and adhesive force. In vivo inhalation of PAE-CD-MOF dry powder inhaler by rat demonstrated rapid absorption (t of 4.0 min) and high absolute bioavailability (71%) of PAE, highlighting significant improvements in absorption and bioavailability of PAE when compared with oral administration (C and absolute bioavailability increased 6.5 and 9.3 folds, respectively). Results of in vivo experiments were consistent with cellular permeability studies (after loading into CD-MOF, the permeability of PAE improved about 5 folds in comparison to the pure PAE). Finally, the efficacy of inhaled PAE for ALI was validated by histopathological examination and via reduced levels of inflammatory factors observed in rat plasma. Overall, targeted pulmonary delivery of paeonol by inhaled PAE-CD-MOF particles appears to be promising method of delivery for treatment of ALI.
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http://dx.doi.org/10.1016/j.ijpharm.2020.119649DOI Listing
September 2020

Emerging well-tailored nanoparticulate delivery system based on in situ regulation of the protein corona.

J Control Release 2020 04 10;320:1-18. Epub 2020 Jan 10.

Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, China.

The protein corona significantly changes the nanoparticle (NP) identity both physicochemically and biologically, and in situ regulation of specific plasma protein adsorption on NP surfaces has emerged as a promising strategy for disease-targeting therapy. In the past decade, great progress in protein corona regulation has been achieved via surface chemistry-based nanomedicine development. This review first outlines the latest advances in bio-nano interactions, with special attention to factors that influence the protein corona, including NP physicochemical properties, the biological environment and the duration time. Second, NP surface chemistry strategies designed to inhibit and regulate protein corona formation are highlighted, with special emphasis on albumin, transferrin, apolipoprotein (apo) E, vascular endothelial growth factor (VEGF) and retinol binding protein 4 (RBP4). Finally, the current techniques used to characterize the protein corona are briefly discussed.
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http://dx.doi.org/10.1016/j.jconrel.2020.01.007DOI Listing
April 2020

Allyl isothiocyanate treatment alleviates chronic obstructive pulmonary disease through the Nrf2-Notch1 signaling and upregulation of MRP1.

Life Sci 2020 Feb 9;243:117291. Epub 2020 Jan 9.

School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, Anhui, China; Anhui Province Key Laboratory of Chinese Medicinal Formula, Hefei 230012, Anhui, China. Electronic address:

Aims: Chronic obstructive pulmonary disease (COPD) is a disease with high morbidity and mortality worldwide, which can cause serious social and economic burdens. Allyl isothiocyanate (AITC) is one of the most common natural isothiocyanates and has been shown to have anti-inflammatory and antioxidant biological activities. The purpose of this study was to investigate whether AITC regulated Multidrug resistance-associated protein 1 (MRP1), reactive oxide species (ROS) and reduced glutathione (GSH) levels via Nrf2 and Notch1 signaling pathways to treat COPD and whether there was an interaction between these two pathways.

Main Methods: Lung function indexes and histopathological changes in mice were determined by lung function instrument and HE staining, respectively. The protein expression was analyzed using immunohistochemistry and Western blotting. The mRNA expression was measured by RT-PCR in human bronchial epithelial cell line 16HBE. The contents of ROS, GSH and GSSG were detected by kits in 16HBE cells.

Key Findings: The protein expression of Notch1, Hes1, MRP1, Nrf2, and HO-1 in lung tissues of WT mice and untransfected cells were significantly down-regulated in COPD, then significantly ameliorated in treatment groups. The protein expression of MRP1, Notch1 and Hes1 in lung tissues of Nrf2 mice were markedly reduced. There was a significant reduction in expression of Nrf2, HO-1 and MRP1 in si-Notch1 transfected cells. Pretreatment with AITC markedly improved oxidative stress and GSH-redox disorder in COPD.

Significance: Our study demonstrates that there is a potential interaction between Nrf2 and Notch1 signaling pathways during treatment of COPD.
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http://dx.doi.org/10.1016/j.lfs.2020.117291DOI Listing
February 2020

Metabolic profiling of chronic obstructive pulmonary disease model rats and the interventional effects of HuaTanJiangQi decoction using UHPLC-Q-TOF/MS.

J Pharm Biomed Anal 2020 Feb 24;180:113078. Epub 2019 Dec 24.

The College of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui, 230031, China. Electronic address:

The occurrence of chronic obstructive pulmonary disease (COPD) will lead to physiological and pathological variations and endogenous metabolic disorders. A traditional Chinese medicine formula, HuaTanJiangQi decoction (HTJQ), exhibits an unambiguous therapeutic effect on COPD in China. Nevertheless, the mechanism of its therapeutic effect on COPD is not clear. With this purpose, pulmonary function, histopathological and the inflammatory factors in bronchoalveolar lavage fluid (BALF) in rats model of COPD were investigated. Then, ultra high-performance liquid chromatography quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) analysis and multivariate statistical analysis were used to further reveal the mechanism of HTJQ therapeutic effect on COPD via metabolomics study. The results showed that the characteristics of lung tissues were significantly reversed, the concentration of LTB4 and LTC4 were gradually decreased, and the lung function began to recover after HTJQ treatment. These typical indicators of COPD in HTJQ intervention group were reversed similar to the control group, suggested that HTJQ has a therapeutic effect on COPD. Moreover, 32 dysregulated metabolites, including Thromboxane a2, Sphingosine 1-phosphate, PC(18:2(9Z,12Z)/18:1(11Z)), Leukotriene B4, Glutathione, Arachidonic acid, Sphingosylphosphocholine acid, N-Acetyl-leukotriene e4, Lysopc(18:1(11Z)), L-Cysteine, and Guanosine diphosphate. All the altered metabolites were associated with the onset and development of COPD, and involved in glycerophospholipid metabolism, sphingolipid metabolism, glutathione metabolism, and arachidonic acid metabolism, which were significantly changed in rats model with COPD. Generally, these findings provide a systematic view of metabolic changes linked to the onset and development of COPD, also indicated that HTJQ could provide satisfactory therapeutic effects on COPD and metabolomics study can be utilized to further understand the molecular mechanisms.
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http://dx.doi.org/10.1016/j.jpba.2019.113078DOI Listing
February 2020

Allyl isothiocyanate may reverse the expression of MRP1 in COPD rats via the Notch1 signaling pathway.

Arch Pharm Res 2019 Nov 30;42(11):1000-1011. Epub 2019 Sep 30.

Anhui University of Chinese Medicine, Hefei, 230031, Anhui, People's Republic of China.

In the present study, the roles of AITC in up-regulating the MRP1 expression and its relationship with the activation of the Notch1 signaling pathway were investigated by combining the in vivo and in vitro experiments. AITC was administered to the COPD model rats and normal rats to explore the association between Notch1 and MRP1. The human bronchial epithelial cells were treated with DAPT, the Notch1 signaling pathway inhibitor, to verify the effect of Notch1 on the expression of AITC-induced MRP1. Compared with the control group, the expressions of Notch1, Hes1 (the target gene of Notch1) and MRP1 in the lung tissue of the COPD model group were significantly inhibited. In contrast to the COPD model group, the expressions of MRP1, Hes1 and Notch1 dramatically up-regulated following the treatment with Low/High doses of AITC. The expression of MRP1 in the 16 HBE cells was down-regulated by the inhibition of Notch in a DAPT concentration-dependent manner. Additionally, the AITC-induced up-regulation of the MRP1 expression was markedly impaired following the inhibition of Notch1. The above results indicated that the pulmonary function and the expression of MRP1 in COPD rats could be improved by AITC, which was partly dependent on the Notch1 signaling pathway. Therefore, targeting the Notch signaling pathway may present as an effective therapeutic strategy for COPD treatment.
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http://dx.doi.org/10.1007/s12272-019-01183-4DOI Listing
November 2019

[Qibai Pingfei capsule alleviates inflammation and oxidative stress in a chronic obstructive pulmonary disease rat model with syndromes of Qi deficiency and phlegm and blood stasis by regulating the SIRT1/FoxO3a pathway].

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2019 Feb;35(2):115-120

College of Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230012, China.

Objective To explore the effect of Qibai Pingfei capsule (QPC) on the inflammation and oxidative stress in a chronic obstructive pulmonary disease (COPD) rat models with the syndromes of qi deficiency and phlegm and blood stasis by regulating the SIRT1/FoxO3a pathway. Methods A total of 80 male SD rats were randomly divided into 4 groups with 20 animals in each group: a non-diseased group, a non-treated diseased group, a diseased group treated with QPC, and a diseased group treated with placebo. The COPD rat models with the syndromes of qi deficiency and phlegm and blood stasis were then developed with established protocols. After the corresponding treatments, the serum levels of superoxide dismutase (SOD), malondialdehyde (MDA), interleukine 1β (IL-1β), and IL-2 were determined by ELISA; the protein levels of SIRT1 and FoxO3a were quantified by Western blot analysis; the mRNA levels of the SIRT1 and FoxO3a genes were also measured by real-time quantitative PCR. Results First of all, compared with the non-diseased group, the serum levels of MDA, IL-1β, and IL-2 were elevated in the diseased group, while the level of SOD was reduced. Both mRNA and protein levels of SIRT1 decreased, while the levels of FoxO3a increased in the lung tissues of the diseased group. Compared with the diseased group treated with placebo, the diseased group treated with QPC had reduced serum levels of MDA, IL-1β and IL-2, elevated SOD, increased mRNA and protein levels of SIRT1 and decreased levels of FoxO3a, thereby restoring their levels partially under the disease state. Conclusion QPC can alleviate inflammation and oxidative stress of COPD rats with syndrome of qi deficiency and phlegm and blood stasis effectively, potentially through regulating the expression level of the SIRT1/FoxO3a pathway.
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February 2019

Long Noncoding RNAs in the Regulation of Oxidative Stress.

Oxid Med Cell Longev 2019 17;2019:1318795. Epub 2019 Feb 17.

Institutes of Integrative Medicine, Fudan University, Shanghai, China.

Oxidative stress takes responsibility for various diseases, such as chronic obstructive pulmonary disease (COPD), Alzheimer's disease (AD), and cardiovascular disease; nevertheless, there is still a lack of specific biomarkers for the guidance of diagnosis and treatment of oxidative stress-related diseases. In recent years, growing studies have documented that oxidative stress has crucial correlations with long noncoding RNAs (lncRNAs), which have been identified as important transcriptions involving the process of oxidative stress, inflammation, etc. and been regarded as the potential specific biomarkers. In this paper, we review links between oxidative stress and lncRNAs, highlight lncRNAs that refer to oxidative stress, and conclude that lncRNAs have played a negative or positive role in the oxidation/antioxidant system, which may be helpful for the further investigation of specific biomarkers of oxidative stress-related diseases.
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http://dx.doi.org/10.1155/2019/1318795DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6398004PMC
April 2019

Untargeted metabolomics profiles delineate metabolic alterations in mouse plasma during lung carcinoma development using UPLC-QTOF/MS in MS mode.

R Soc Open Sci 2018 Sep 19;5(9):181143. Epub 2018 Sep 19.

Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui Province Key Laboratory of R&D of Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230038, People's Republic of China.

In this work, an untargeted metabolomic method based on ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) in MS (E represents collision energy) mode was exploited to determine the dynamic metabolic alterations in the plasma of male C57BL/6 mice during the onset and development of lung carcinoma. Plasma samples were collected from control and model mice (male C57BL/6 mice experimentally inoculated with the lung carcinoma cells) at 7 and 14 days post-inoculation (DPI). As a result, 15 dysregulated metabolites, including cholesterol sulphate, tiglylcarnitine, 1-palmitoylglycerophosphoinositol, 2-stearoylglycerophosphoinositol, stearoylcarnitine, PC(20:2(11Z,14Z)/16:0), PC(22:4(7Z,10Z,13Z,16Z)/14:0), PC(22:5(7Z,10Z,13Z,16Z,19Z)/14:0), PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/16:0), 12,20-Dioxo-leukotriene B4, sphingosine 1-phosphate(d19:1-P), sphingomyelin(d18:0/16:1(9Z)), lysoPC(16:0), lysoPC(18:0) and lysoPC(20:4(5Z,8Z,11Z,14Z)), were identified in the plasma of model mice with xenografts at both 7 and 14 DPI. All the altered metabolites associated with the onset and development of lung carcinoma were involved in the metabolism of glycerophospholipid, fatty acid, sphingolipid and arachidonic acid. The feasible utility of these endogenous biomarkers as potential diagnostic indicators was validated through receiver operating characteristic curve analysis. Collectively, these findings provide a systematic view of metabolic changes linked to the onset and development of lung carcinoma.
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http://dx.doi.org/10.1098/rsos.181143DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6170569PMC
September 2018

De novo sequencing and transcriptome assembly of Arisaema heterophyllum Blume and identification of genes involved in isoflavonoid biosynthesis.

Sci Rep 2018 12 5;8(1):17643. Epub 2018 Dec 5.

Anhui University of Chinese Medicine and Anhui Academy of Chinese Medicine, Hefei, 230038, China.

Arisaema heterophyllum Blume (AhBl) is one of the valued medicinal plants. However, its genetic information is limited, which impedes further studies of this valuable resource. To investigate the genes involved in the isoflavonoid biosynthesis, we deeply performed transcriptome sequencing for AhBl. An average of 10.98 Gb clean reads were obtained based on root, tuber and leaf tissues, and 109,937 unigenes were yielded after de novo assembly. In total, 72,287 of those unigenes were annotated in at least one public database. The numbers of expressed unigenes in each tissue were 35,686, 43,363 and 47,783, respectively. The overall expression levels of transcripts in leaf were higher than those in root and tuber. Differentially expressed genes analysis indicated that a total of 12,448 shared unigenes were detected in all three tissues, 10,215 of which were higher expressed in tuber than that in root and leaf. Besides, 87 candidate unigenes that encode for enzymes involved in biosynthesis of isoflavonoid were identified and analyzed, and some key enzyme genes were experimentally validated by quantitative Real-Time PCR (qRT-PCR). This study provides a unique dataset for the systematic analysis of AhBl functional genes and expression characteristics, and facilitates the future study of the pharmacological mechanism of AhBl.
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http://dx.doi.org/10.1038/s41598-018-35664-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6281570PMC
December 2018

Intervention effect of Qi-Yu-San-Long Decoction on Lewis lung carcinoma in C57BL/6 mice: Insights from UPLC-QTOF/MS-based metabolic profiling.

J Chromatogr B Analyt Technol Biomed Life Sci 2018 Dec 17;1102-1103:23-33. Epub 2018 Oct 17.

Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui Province Key Laboratory of R&D of Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230038, China; Institute of Pharmaceutics, Anhui Academy of Chinese Medicine, Hefei 230012, China; National Key Disciplines of Lung Disease of Anhui University of Chinese Medicine, State Administration of Traditional Chinese Medicine, Hefei 230038, China. Electronic address:

Qi-Yu-San-Long Decoction (QYSLD) has been used to treat lung carcinoma for over twenty years in clinical practices, and its curative effect is considered credible. However, the therapeutic mechanism of this effect has not been thoroughly elucidated to date. In this study, a MTT dye reduction assay and DAPI staining were first used to evaluate the cell viability and apoptosis of A549 cells with and without QYSLD-treatment, respectively. The weight/volume of Lewis lung carcinoma (LLC) sarcoma was used to assess the therapeutic effect of QYSLD on LLC mice. Second, an UPLC-QTOF/MS-based untargeted metabolomics method was employed to identify and relatively quantify functional metabolites that were responsible for the intervention effect of QYSLD on LLC. As a result, the MTT dye reduction assay and DAPI staining demonstrated that QYSLD could inhibit the proliferation and induce the apoptosis of A549 cells. The weight/volume test of LLC sarcoma showed that QYSLD could restrain the development of LLC. Next, 21 potential biomarkers that could contribute to the curative mechanism of QYSLD on LLC were screened by the untargeted metabolomics method. The down-regulated metabolites induced by QYSLD included PC(16:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z)), PC(20:2(11Z,14Z)/16:0), PC(22:4(7Z,10Z,13Z,16Z)/14:0), PC(22:5(7Z,10Z,13Z,16Z,19Z)/14:0), arachidonic acid, gamma-glutamylisoleucine, cholesterol sulfate, CL (8:0/10:0/11:0/a-13:0) and CDP-DG (16:0/18:1(11Z)). The up-regulated metabolites were LysoPC(16:0), LysoPC(18:0), LysoPE(18:2(9Z,12Z)/0:0), LysoPE(22:0/0:0), LysoPE(22:1(13Z)/0:0), LysoPE(22:2(13Z,16Z)/0:0), triglylcarnitine, 1‑arachidonoylglycerophosphoinositol, 1‑palmitoylglycerophosphoinositol, 2‑stearoylglycerophosphoinositol, sphingosine 1‑phosphate(d19:1-P) and SM(d18:0/16:1(9Z)). The metabolic pathway analysis revealed that the potential biomarkers were primarily involved in glycerophospholipid metabolism, sphingolipid metabolism, steroid hormone biosynthesis, fatty acid degradation and arachidonic acid metabolism. This study demonstrated that QYSLD has a good antitumor effect and that a UPLC-QTOF/MS-based untargeted metabolomics method is a promising means of elucidating the intervention mechanism of traditional Chinese medicine formulas.
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http://dx.doi.org/10.1016/j.jchromb.2018.10.013DOI Listing
December 2018

Pingchuanning decoction attenuates airway inflammation by suppressing autophagy via phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway in rat models of asthma.

J Cell Biochem 2019 03 27;120(3):3833-3844. Epub 2018 Sep 27.

Institute of Traditional Chinese Medicine of Respiratory Disease Prevention, Anhui University of Traditional Chinese Medicine, Hefei, Anhui, China.

Background: Pingchuanning decoction is a well-known traditional Chinese medicine for the treatment of airway inflammatory diseases, including asthma. However, the potential mechanism by which Pingchuanning decoction contributes to the amelioration of airway inflammation remains unknown.

Methods: A rat model of asthma was well established by inducing ovalbumin. Lipopolysaccharide-stimulated rat tracheal epithelial (RTE) cells were used as cellular model. Lung histopathology and goblet cell hyperplasia were assessed by hematoxylin-eosin (HE) and periodic acid Schiff staining, respectively. Total inflammatory cells count and RTE cell apoptosis were analyzed by flow cytometry. The autophagic activities were evaluated by immunohistochemical and immunofluorescence analysis and Western blot analysis of autophagy-related proteins. We also detected the effects of Pingchuanning decoction on phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) and high-mobility group box 1 (HMGB1)-mediated toll-like receptor 4 (TLR4)/NF-κB pathways-related proteins and inflammatory cytokines using the Western blot analysis and enzyme-linked immunosorbent assay.

Results: Pingchuanning decoction effectively attenuated pulmonary pathology and autophagy. Treatment with Pingchuanning decoction activated PI3K/Akt/mTOR pathway and inhibited HMGB1/TLR4/NF-κB pathway, which could be overturned by LY294002, a PI3K antagonist, or rapamycin (Rapa), an autophagy inducer.

Conclusion: Pingchuanning decoction exerted a therapeutic effect on asthma by inhibiting autophagy via PI3K/Akt /mTOR signaling pathway.
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http://dx.doi.org/10.1002/jcb.27665DOI Listing
March 2019

Effects of Qibaipingfei capsules on pulmonary vascular relaxation through KATP channel activation by the NO/cGMP signaling pathway.

Pak J Pharm Sci 2018 Mar;31(2):545-552

Hubei University of Chinese Medicine, Wuhan, China / The Key Discipline of SATCM-TCM Lung Disease, Anhui University of Chinese Medicine, Hefei, China.

This research explores the effects of Qibaipingfei (QBPF) capsules on pulmonary vascular relaxation in vitro and the relationship of the ATP-sensitive K (KATP) channel and nitric oxide (NO) pathway. Vasodilator effects of QBPF (0.125-2 g/kg) on rat pulmonary artery rings were observed using a multi-wire myograph system. The maximum relaxation (E) of QBPF was detected following treatment involving endothelial denudation, Nω-nitro-L-arginine methyl ester (L-NAME), 1H-[1,2,4] oxadiazolo[4,3-α]quinoxalin-1-one (ODQ), or glyburide (GLYB). Furthermore, rat models of phlegm and blood stasis syndrome combined with chronic obstructive pulmonary disease (COPD) were established using compound factors. KIR6.1 and SUR2B protein expression was analyzed by western blotting. After 9,11-dideoxy-11-α],9-α]-epoxy-methanoprostaglandinF (U46619) was used to pre-constrict endothelium-intact pulmonary artery rings, QBPF induced the effects of concentration-dependent relaxation at a concentration for 50% of maximal effect (EC) of 0.56 g/L and E of 84.30% ± 6.27%. After the endothelium was denuded, the vasodilator effects reduced significantly (P<0.01). QBPF-induced relaxation was inhibited by L-NAME, ODQ, and GLYB (P<0.01). The vasodilator effect was also attenuated in the model group (E=62.63%±10.02, EC = 0.72 g/L, P<0.01). In comparison with expression in the control group, SUR2B protein expression was down-regulated in the model group (P<0.01) but no significant difference was detected in KIR6.1 protein expression between the groups (P>0.05). QBPF and nicorandil (Nic) treatment up-regulated SUR2B K channel expression (P<0.05). QBPF induces endothelial-dependent relaxation in pulmonary artery rings in vitro, through a mechanism that potentially activates the K channel in pulmonary vascular smooth muscles via the NO-cyclic GMP (cGMP)-dependent pathway.
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March 2018

Huaier Granule extract inhibit the proliferation and metastasis of lung cancer cells through down-regulation of MTDH, JAK2/STAT3 and MAPK signaling pathways.

Biomed Pharmacother 2018 May 22;101:311-321. Epub 2018 Mar 22.

Anhui University of Chinese Medicine, Hefei, People's Republic of China. Electronic address:

Although the effect of Huaier has been widely studied, its role and its molecular mechanism in lung cancer are not clear. In this study, we explored the inhibitory effect of Huaier on lung cancer cells and its molecular mechanism. The cell viability, migration and invasion were analyzed by CCK-8 and BrdU cell proliferation assay kits, Transwell and colony forming assay. The cell cycle and apoptosis were analyzed by flow cytometry. The experimental results showed that the viability, migration and invasion of A549 and NCI-H1650 cells were inhibited by Huaier in a dose and time-dependent manner. Huaier induced cell apoptosis and the cells were blocked in the S phase to inhibit cell proliferation. Western blotting results showed that Huaier inhibited the expression of MTDH and increased the proportion of Bax/Bcl-2, it could also promote the expression of Cleaved Caspase-3 and increase the activity of Caspase-3, promote cell apoptosis and inhibit cell proliferation. Huaier inhibited the metastasis and invasion of lung cancer cells by inhibiting the expression levels of EMT related proteins, it also inhibited the expression of JAK2/STAT3 and MAPK signaling pathways. Therefore, our results showed that Huaier may inhibit the proliferation and metastasis of lung cancer cells through multiple targets, it had the potential for treatment of lung cancer.
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http://dx.doi.org/10.1016/j.biopha.2018.02.028DOI Listing
May 2018

Effect of Liuweibuqi Capsules in Pulmonary Alveolar Epithelial Cells and COPD Through JAK/STAT Pathway.

Cell Physiol Biochem 2017 27;43(2):743-756. Epub 2017 Sep 27.

Department of Traditional Chinese Medicine, The First Affiliated Hospital, Anhui Medical University, Hefei, China.

Background/aims: To evaluate the effect of Liuweibuqi capsules on chronic obstructive pulmonary disease (COPD) through the JAK/STAT pathway.

Methods: Lung function was measured with a spirometer. Changes in lung histology were observed using H&E staining. Cigarette smoke extract combined with lipopolysaccharide (CSE+LPS) was used to establish the cellular COPD model. Cytokine levels were determined using ELISA, and changes in the JAK/STAT pathway were evaluated using western blotting. The CCK8 method and flow cytometry were used to measure cell viability and apoptosis, respectively.

Results: Liuweibuqi capsules reduced the damage in the lung tissues and the loss of lung function in the COPD rats. Additionally, the levels of interleukin (IL)-1β, interferon γ (IFNγ), IL-6, tumor necrosis factor (TNF)-α and transforming growth factor (TGF)-β1 were higher, whereas IL-10 was lower in the model control (MC) and CSE+LPS groups than in the normal group. The phosphorylation levels of JAK1, JAK2, STAT1, STAT3 and STAT5 were higher and the levels of SOCS1 and SOCS3 were lower in the MC group and CSE+LPS group compared with the normal group. After Liuweibuqi capsule treatment, the expression of inflammatory cytokines and elements of the JAK/STAT pathway were lower. In addition, over-expression of STAT3 blocked the effects of the Liuweibuqi capsules on the release of inflammatory cytokines, cell viability and apoptosis.

Conclusion: Our findings suggested that Liuweibuqi capsule might effectively ameliorate the progression of COPD via the JAK/STAT pathway.
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http://dx.doi.org/10.1159/000481558DOI Listing
October 2017

Effect of Liuweibuqi capsules on the balance between MMP-9 and TIMP1 and viability of alveolar macrophages in COPD.

Biosci Rep 2017 Oct 19;37(5). Epub 2017 Sep 19.

Department of Traditional Chinese Medicine, The First Affiliated Hospital, Anhui Medical University, Hefei 230022, China.

The present study aims to investigate the effect of Liuweibuqi (LWBQ) capsules on the expression of matrix metalloproteinase (MMP)-9 and TIMP1 and cell viability of alveolar macrophages (AMs) in chronic obstructive pulmonary disease (COPD). Rats were randomly divided into normal control (NC) group, model control (MC) group, Jinshuibao (JSB) group, spleen aminopeptidase (PAT) group, and low dose of LWBQ (LWBQ low), mid dose of LWBQ (LWBQ mid), and high dose of LWBQ (LWBQ high) group (=10). Lung function was measured with a spirometer. Serum cytokines including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected using ELISA. The expressions of MMP-9 and TIMP1 were detected by quantitative real-time PCR (qRT-PCR) and Western blot. MTT assay and flow cytometry were used to measure cell viability and apoptosis. Compared with the NC group, body weight and lung function were reduced in the MC group. In addition, the serum levels of IL-6 and TNF-α were higher in the MC group than those in the NC group. The expression of MMP-9 protein in the AMs from rats was higher, and TIMP1 protein was lower in the MC group compared with the NC group. After LWBQ capsules treatment, compared with the MC group, the expression of inflammatory cytokines and MMP-9 were lower and TIMP1 was higher. Moreover, after LWBQ-medicated serum treatment, the release of inflammatory cytokines was reduced from AMs. Besides, LWBQ-medicated serum decreased the expression of MMP-9 and increased the expression of TIMP1 and cell viability compared with those in MC group. In conclusion, LWBQ capsules can inhibit the release of inflammatory cytokines, promote cell viability in AMs, and regulate the expression of MMP-9 and TIMP1.
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http://dx.doi.org/10.1042/BSR20170880DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5603752PMC
October 2017

Qibai Pingfei capsule medicated serum inhibits the proliferation of hypoxia-induced pulmonary arterial smooth muscle cells via the Ca2+/calcineurin/nuclear factor of activated T-cells 3 pathway.

J Tradit Chin Med 2017 Aug;37(4):466-474

Anhui Academy of Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230038, China.

Objective: To observe the effect of Qibai Pingfei capsule (QBPF) medicated serum on the proliferation of rat pulmonary arterial smooth muscle cells (PASMCs) under hypoxia conditions and to investigate its key molecular effects on the Ca2+/calcineurin/nuclear factor of activated T-cells 3 (NFATc3) signaling pathway.

Methods: QBPF was provided to rats via continuous gavage for 10 days. Primary rat PASMCs were cultured using the direct adherent culture method. Methyl thiazolyl tetrazolium assay was used to evaluate the effect of QBPF on PASMCs proliferation under hypoxia conditions. Laser scanning confocal microscopy was used to detect changes in intracellular free calcium ([Ca2+]i) in PASMC-loaded Fluo-3-AM. Real-time quantitative polymerase chain reaction and western blot were used to detect the transcription and protein expression levels of calcineurin and NFATc3 genes in PASMCs.

Results: Compared with normoxia conditions, PASMCs proliferated at 12, 24, 48, and 72 h under hypoxia conditions. QBPF at concentrations of 5%, 10%, and 20% could inhibit hypoxia-induced PASMC proliferation to different degrees. The inhibitory effect was most significant in the 20% QBPF group under 24 h hypoxia conditions. The concentration of [Ca2+]i in PASMCs under hypoxia was increased and [Ca2+]i was significantly decreased when co-incubated with QBPF at 5%, 10%, and 20%. Compared with normoxia conditions, the mRNA and protein expression levels of calcineurin and NFATc3 in PASMCs induced by hypoxia were up-regulated. QBPF application significantly down-regulated mRNA and protein expression levels of calcineurin and NFATc3 in PASMCs.

Conclusion: QBPF can effectively inhibit hypoxia-induced proliferation of PASMCs through down-regulation of key molecular expression via the Ca2+/calcineurin/NFATc3 pathway.
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August 2017

[Qiyusanlong decoction inhibits the level of PD-1/PD-L1 in mice bearing Lewis lung carcinoma].

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2016 Jun;32(6):770-4

Anhui University of Chinese Medicine, Hefei 230038; Institute of Chinese Medicine for Respiratory Diseases, Anhui Provincial Academy of Chinese Medical Sciences, Hefei 230038, China. *Corresponding author, E-mail:

Objective To investigate the anti-tumor effect of Qiyusanlong decoction (QYSL) in the mice with Lewis lung cancer (LLC) and its effect on the expression of programmed death 1 and programmed death ligand 1 (PD-1/PD-L1). Methods The model of lung cancer subcutaneous allograft was established using LLC cells. The model mice were randomly divided into six groups: a model group, a chemotherapy group, three QYSL groups of high, middle and low doses, and a combined group, each group containing 8 mice. On the 11th day, the low-, middle- and high-dose QYSL groups were respectively given intragastric administration of 20.12, 40.24, 80.48 g/(kg.d) QYSL; the chemotherapy group were intraperitoneally injected with 0.4 mL cisplatin; the combined group were administrated with cisplatin and high-dose QYSL; the model group were administrated with the same amount of normal saline, once a day for 10 days. Tumor volume was examined and tumor growth curve was drawn. Tumors were weighed and tumor inhibition rates were calculated. The expressions of PD-1, PD-L1 mRNA and protein in spleens and tumor tissues of mice were detected by real-time quantitative PCR and Western blotting, respectively. Results Compared with the model group, the high-dose QYSL group could inhibit tumor growth with tumor inhibition rate being 23.86%; the tumor inhibition rate between the combined group and the chemotherapy group was equal. The high-dose QYSL could significantly decrease the expression of PD-1 mRNA and protein in spleen and inhibited the expression of PD-L1 mRNA and protein in tumor. The levels of PD-1 and PD-L1 protein in the combined group were obviously lower than those in the chemotherapy group, but the interaction effect of the other indicators had no statistical significance. Conclusion QYSL can moderately inhibit the growth of the transplanted tumor by decreasing PD-1/PD-L1 level.
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June 2016

Ligustrazine improves blood circulation by suppressing Platelet activation in a rat model of allergic asthma.

Environ Toxicol Pharmacol 2016 Jul 15;45:334-9. Epub 2016 Jun 15.

Anhui University of Chinese Medicine, No. 103 Meishan Road, Hefei 230012, Anhui, China. Electronic address:

Chuan-xiong (Ligusticum wallichii) is a traditional herbal medicine in Eastern Asia, but the effect of its active component ligustrazine remains unclear. We explored its effect and possible mechanism in a well-characterized rat model of allergic asthma. Ligustrazine suppressed bronchial hyper-responsiveness to methacholine, and suppressed lung inflammation in asthmatic rats. Ligustrazine exhibited potent immuno-modulatory and anti-inflammatory effects: it suppressed lymphocyte and eosinophil mobilization, and reduced cytokine IL-5 and IL-13 production significantly in lung tissues from asthmatic rats (P<0.05). Further histological examinations clearly demonstrated that ligustrazine improved blood circulation and ameliorated platelet activation, aggregation and adhesion, which induced sustained infiltration and activation of various inflammatory cells, including lymphocytes and eosinophils, followed by synthesis and release of a variety of pro-inflammatory mediators and cytokines. The present study suggests that ligustrazine is a potent agent for the treatment of allergic asthma due to its strong anti-inflammatory and immuno-modulatory properties.
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http://dx.doi.org/10.1016/j.etap.2016.06.016DOI Listing
July 2016

[Qibaipingfei Capsule down-regulate the levels of calcineurin and nuclear factor of activated T-cells isoform c3 (NFATc3) in chronic obstructive pulmonary disease].

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2015 Nov;31(11):1497-501

Key Discipline of SATCM-TCM Lung Disease, Anhui University of Chinese Medicine, Hefei 230031, China.

Objective: To investigate the effect of Qibaipingfei Capsule (QPC) on the expressions of calcineurin (CaN) and nuclear factor of activated T-cells isoform c3 (NFATc3) of rat models with phlegm and blood stasis syndrome of chronic obstructive pulmonary disease (COPD), and to explore the possible mechanism underlying the intervention of QPC in pulmonary vascular remodeling of COPD.

Methods: Sixty male Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, a positive group of nifedipine, a high dose group, a middle dose group and a low dose group of QPC. The rat models with phlegm and blood stasis syndrome of COPD were established by compound methods of forced swimming, smoking and hypoxia. Then the pulmonary function and the pathological alterations of pulmonary vessels were observed. Furthermore, the mRNA and protein levels of CaN and NFATc3 in the lung tissues were determined by real-time quantitative PCR and Western blot analysis.

Results: Compared with the normal group, the forced expiratory volume at 0.3 second (FEV0.3), the forced vital capacity (FVC) and FEV0.3/FVC in the model group were significantly reduced, but compared with the model group, the values mentioned above were restored to different extents in the groups of nifedipine and QPC. The lung tissues of the model group showed the thickening of pulmonary vascular wall and the formation of compensating emphysema. The above pathological changes were relieved in all the treatment groups. Compared with the normal group, the expressions of CaN and NFATc3 in the model group were significantly up-regulated in transcription and translation levels. Compared with the model group, these expressions were down-regulated to various degrees in all the treatment groups.

Conclusion: QPC can decrease the levels of CaN and NFATc3 in the lung tissues of COPD.
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November 2015

Upregulation of Multidrug Resistance-Associated Protein 1 by Allyl Isothiocyanate in Human Bronchial Epithelial Cell: Involvement of c-Jun N-Terminal Kinase Signaling Pathway.

Oxid Med Cell Longev 2015 27;2015:903782. Epub 2015 Jul 27.

Anhui University of Chinese Medicine, Hefei, Anhui 230038, China.

Multidrug resistance-associated protein 1 (MRP1) plays a protective role in the etiology and progression of chronic obstructive pulmonary disease (COPD) which results from oxidative stress and inflammation of lung injury. The lower functional MRP1 activity is related to COPD development. Our previous study showed that Allyl isothiocyanate (AITC) induced the expression and activity of MRP1 in a dose-dependent manner. However, which signaling pathway contributes to the upregulation of MRP1 by AITC is unclear. In this study, signaling pathway specific inhibitors were used to examine the mechanism of AITC. We found that JNK inhibitor SP600125 treatment decreased MRP1 mRNA expression in 16HBE14o- cells. But the ERK inhibitor U0126 or PI3K/Akt inhibitor LY294002 produced no obvious effect. The AITC-induced increase of MRP1 mRNA expression was abolished by cotreatment of SP600125, while it was not obviously affected by U0126 or LY294002. Furthermore, AITC acivates the JNK signaling pathway in 16HBE14o- cells. Finally, we found that JNK pathway mediated the upregulation of AITC-induced expression and function of MRP1. Taken together, our results indicated that AITC increased the expression and the activity of MRP1 via a JNK-dependent pathway. ERK and PI3K signaling pathway were not involved in the expression of MRP1 mRNA.
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http://dx.doi.org/10.1155/2015/903782DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4530297PMC
May 2016

[Role of Foxp3/Treg and RORγt/Th17 cell imbalance in rat model of chronic obstructive pulmonary disease].

Zhonghua Wei Zhong Bing Ji Jiu Yi Xue 2014 Dec;26(12):860-4

Department of Chinese Medicine, the First Affiliated Hospital of Anhui Medical University, Hefei 230022, Anhui, China, Corresponding author: Li Zegeng, Department of Respiratory Diseases, the First Affiliated Hospital of Anhui University of Traditional Medicine, Hefei 230031, Anhui, China,

Objective: To observe the changes in forkhead/winged helix transcription factor p3(Foxp3), regulatory T cells (Treg), retinoid-related orphan receptor gamma (RORγt) in rat model of chronic obstructive pulmonary disease (COPD).

Methods: Twenty Sprague-Dawley (SD) rats were randomly divided into normal control group and COPD model group, with 10 rats in each group. The COPD model was reproduced by smoke inhalation and tracheal instillation of lipopolysaccharide (LPS), and no such treatment was conducted in normal control group. Twenty-eight days after the model reproduction, the pulmonary function was determined, the pathological changes of lung tissue were observed with haematoxylin-eosin (HE) staining, interleukins (IL-6, IL-10) in serum were detected by enzyme-linked immunosorbent assay (ELISA), CD4⁺ CD25⁺ Foxp3⁺ Treg of peripheral blood was determined by flow cytometry, and the expressions of Foxp3, RORγt, IL-17 protein in lung tissue were assayed by Western Blot.

Results: Under light microscope, significal interstitial infiltration of inflammatory cells was found in alveoli and interstitial tissue of the lung, and destruction of alveolar tissue, alveolar wall thinning, and even rupture to fuse into bullae, and bleeding into alveoli in different degress could be observed. Compared with the normal control group, forced vital capacity (FVC), forced expiratory volume in 0.3 second (FEV0.3), FEV0.3/FVC, peak expiratory flow (PEF) in model group were significantly decreased [FVC (mL): 8.04 ± 2.03 vs. 9.97 ± 2.14, FEV0.3 (mL): 6.16 ± 2.23 vs. 8.84 ± 2.12, FEV0.3/FVC: 0.70 ± 0.09 vs. 0.85 ± 0.11, PEF (mL/s): 33.56 ± 4.76 vs. 40.14 ± 5.64, P<0.05 or P<0.01]. Serum IL-6 was obviously increased (ng/L: 93.17 ±20.96 vs. 76.28 ± 13.24, P<0.05), IL-10 was significantly decreased (ng/L: 78.62 ± 15.17 vs. 104.34 ± 19.46, P<0.01), and CD4⁺ CD25⁺ FoxP3(+)Treg was significantly diminished [(2.75 ± 0.83)% vs. (4.16 ± 1.14)%, P<0.01] in model group compared with those in the normal control group. The expression of Foxp3 protein in lung tissue in model group was significantly down-regulated compared with that in the normal control group (gray scale: 0.38 ± 0.15 vs. 0.63 ± 0.11, P<0.01), and RORγt and IL-17 protein expressions were significantly up-regulated [RORγt (gray scale): 0.96 ± 0.23 vs. 0.47 ± 0.11, IL-17 (gray scale): 1.02 ± 0.24 vs. 0.34 ± 0.08, both P<0.01]. Correlation analysis showed that FEV0.3 was positively correlated with Foxp3 (r=0.585, P<0.05), and FEV0.3/FVC was negatively correlated with IL-6 and RORγt (r=-0.655, r=-0.607, both P<0.05). PEF was positively correlated with Treg (r=0.573, P<0.05), and negatively correlated with IL-17 (r=-0.198, P<0.05). IL-6 was negatively correlated with Foxp3(r=-0.603, P<0.05),and positively correlated with RORγt (r=0.588, P<0.05). IL-10 was positively correlated with Treg (r=0.573, P<0.05). Treg was positively correlated with Foxp3 (r=0.607, P<0.05), and negatively correlated with IL-17 (r=-0.569, P<0.05). Foxp3 was negatively correlated with RORγt (r=-0.591, P<0.05). RORγt was positively correlated with IL-17 (r=0.578, P<0.05).

Conclusions: There is a relationship among decreased pulmonary function, inflammation and imbalance of Foxp3/Treg and RORγt/Th17 in COPD.
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http://dx.doi.org/10.3760/cma.j.issn.2095-4352.2014.12.003DOI Listing
December 2014

[Imbalance in the expression of Foxp3, T-bet and GATA-3 is correlated with inflammation of chronic obstructive pulmonary disease].

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2014 Jul;30(7):704-7, 712

Department of Respiration, First Affiliated Hospital, Anhui University of Traditional Chinese Medicine, Hefei 230031, China.

Objective: To observe the change of regulatory T cells(Tregs), fork head-like transcription factor 3 (Foxp3), T box expressed in T cells (T-bet) and GATA binding protein 3 (GATA3) in rat models of chronic obstructive pulmonary disease (COPD).

Methods: Thirty rats were randomly divided into control group and model group (n=15 each). The rats of model group were developed by lipopolysaccharide (LPS) and smoking. Interleukin-4 (IL-4) and γ-interferon (IFN-γ) in serum and bronchoalveolar lavage fluid (BALF) were detected 28 days after modeling by ELISA. Peripheral Tregs were detected by flow cytometry. The expressions of Foxp3, T-bet, GATA gene and protein in lung tissue were observed by reverse transcription PCR and Western blotting, respectively.

Results: Compared with the control group, the model group had more serious inflammation in lung tissues, expressed the higher levels of IFN-γ in serum and BALF (P<0.01), T-bet mRNA and protein in lung tissue, and the lower levels of IL-4, CD4⁺ CD25⁺ Treg, CD4⁺ CD25⁺ Foxp3⁺ Treg (P<0.05), Foxp3, GATA-3 mRNA and protein (P<0.01). Correlation analysis showed that there were correlations between T-bet, GATA-3, Foxp3 expressions and IL-4, IFN-γ levels (P<0.05).

Conclusion: There is a relationship between inflammation and imbalance of in the expression of T-bet, GATA-3, Foxp3 in COPD.
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July 2014

[Relationship of reduced lung function with Th1/Th2 polarization, STAT4/6 expression in rats of chronic obstructive pulmonary disease].

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2013 Dec;29(12):1233-6

Graduate Division, Hubei University of Chinese Medicine, Wuhan 430065, China.

Objective: To study the correlations between lung function and Th1/Th2 polarization, signal transducer and activator of transcription (STAT) proteins in rats of chronic obstructive pulmonary disease (COPD).

Methods: Sixty rats were randomly divided into normal control group, sham group and model group. The COPD rat models were developed by lipopolysaccharide (LPS). The changes of pulmonary function were detected at 28 d after modeling. The lung tissues of rats were observed by HE staining, and serum IFN-γ, IL-4, IL-12, and IL-13 were determined by ELISA. The mRNA expressions of IFN-γ and IL-4 were examined by PCR, and STAT4, STAT6 proteins in lung tissues were detected by Western blotting.

Results: Inflammation of lung tissues was more serious, the levels of serum IFN-γ, IL-12 and Th1/Th2, IFN-γ mRNA and STAT4 protein in lung tissues were higher, and lung function, the levels of serum IL-4 and IL-13, IL-4 mRNA and STAT6 protein in lung tissues were lower in model group than those in normal control and sham groups (P < 0.05 or P < 0.01). Correlation analysis showed that there were negative correlations between pulmonary function parameters and IFN-γ, Th1/Th2, STAT4 protein, and positive correlations between pulmonary function parameters and IL-4, IL-13, IL-4 mRNA, STAT6 protein (P < 0.01 or P < 0.05).

Conclusion: The reduced lung function was found related to airway inflammation, cell Th1/Th2 imbalance in COPD. STAT4 and STAT6 involve in the regulation of Th1/Th2 cells. All of the above factors lead to the occurrence of COPD in rats.
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December 2013
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