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ObjectiveTo observe the lung injury of rats caused by PM2.5 induced imbalance of TH17/Treg immune system and the intervention effect of two different TCM treatments.Methods Wistar male rats were randomly divided into normal group, model group, TCM-treated group1 and TCM-treated group 2. PM2.5-induced lung injury model was established by airway instillation. Model group was given normal saline for gavage. TCM- treated group 1 and 2 were given Yupingfeng Powder and Guomin Decoction combined with Zhisou Powder for gavage. The pathological changes of bronchial and lung tissues, the contents of IL-8, IL-10, IL-17, NE, and MUC5AC in serum and BALF were compared, and the expressions of Foxp3 and IL-17 in lung tissue of each group were analyzed.Results Compared with normal group, the contents of IL-8, IL-17, NE and MUC5AC in serum and BALF of model group increased significantly, while IL-10 decreased significantly (P<0.05,P<0.01); the expression of IL-17 increased significantly and the expression of Foxp3 decreased significantly in lung tissue (P<0.01). Compared with model group, the contents of IL-8, IL-17, and NE decreased in TCM-treated group 1 and 2, while the content of BALF IL-10 increased significantly (P<0.05). The content of IL-10 in serum increased significantly in TCM-treated group 2 (P<0.05); the protein expression of IL-17 of lung issue decreased significantly, and the protein expression of Foxp3 increased significantly (P<0.01). The pathological changes were improved significantly.Conclusion PM2.5 can induce lung injury caused by the imbalance of TH17/Treg. Both two treatments can significantly improve the lung injury induced by PM2.5 and the imbalance of TH17/Tregs immune system.
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Objective To explore the mechanism ofJiedu Qingfei Mixture for airway mucus hypersecretion of rat models with chronic obstructive pulmonary disease (COPD).Methods Airway instilling lipopolysaccharide combining fuming method was used to establish COPD models. Forty clean level Wistar strain rats were randomly divided into blank control group, model group,Jiedu Qingfei group, and clarithromycin group. Model group, Jiedu Qingfei group, and clarithromycin group were given normal saline,Jiedu Qingfei Mixture, and clarithromycin by gavage respectively, while the blank control group was raised normally for 30 d. All rats were killed on the 31st day for taking lung tissue (6 rats from each group were chosen randomly). Pathological changes of lung tissue and mucous glands hyperplasia were observed by HE staining method. NE and MUC5AC mRNA expression on lung tissue were detected by RT-PCR method. Protein expressions of NE and MUC5AC on pulmonary tissue and airway epithelium were detected by immunohistochemical method.Results Compared with blank control group, mucous glands hyperplasia on airway epithelium, mRNA expression of NE and MUC5AC in lung tissue, and protein expressions of NE and MUCA5C on airway epithelium in the model group significantly increased (P<0.05,P<0.01). Compared with model group, mucous glands hyperplasia on airway epithelium inJiedu Qingfei group significantly decreased (P<0.01), as same as clarithromycin group;Jiedu Qingfei group showed better effects on down-regulating NE and MUC5AC mRNA expression in lung tissue compared with clarithromycin group. MUC5AC protein expression on airway epithelium inJiedu Qingfei group significantly decreased (P<0.05), as same as clarithromycin group.Jiedu Qingfei group and clarithromycin group had no difference on NE protein expression in airway epithelium compared with model group.Conclusion Jiedu Qingfei group Mixture can reduce airway mucus hypersecretion of COPD by down-regulating MUC5AC expression through neutrophil elastase.
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Objective To observe the effects of Qingjin Huatan Decoction on EGFR/MAPK signaling pathway of airway mucus hypersecretion rats with chronic obstructive pulmonary disease (COPD). Methods Intratracheal instillation of LPS combined with smudging method was used to establish COPD airway mucus hypersecretion rat models. Experimental rats were randomly divided into blank group, model group, Qingjin Huatan Decoction group and clarithromycin group. The blank group was normally fed, while the other three groups were given NS, Qingjin Huatan Decoction, and clarithromycin respectively for gavage, once a day for 30 days. All rats were killed on the 31st day, and pathological changes of lung tissue and mucous glands hyperplasia were observed by HE staining method. The gene expressions of EGFR and MUC5AC in lung tissue were detected by RT-PCR method. The protein expressions of P-EGFR, P-ERK, P-JNK, P-p38 and MUC5AC in pulmonary tissue and airway epithelium were detected by immunohistochemical method. Results Compared with the blank group, mucous glands hyperplasia on airway epithelium, protein expressions of P-EGFR, P-ERK, P-JNK, P-p38 and MUC5AC on airway epithelium significantly increased in the model group (P<0.01); gene expression of MUC5AC of lung tissue increased (P<0.05). Compared with the model group, mucous glands hyperplasia on airway epithelium, P-p38, P-ERK and MUC5AC protein expression on airway epithelium in Qingjin Huatan Decoction group significantly decreased (P<0.05, P<0.01); the protein expression of P-JNK increased significantly (P<0.01). EGFR and MUC5AC mRNA in lung tissue in Qingjin Huatan Decoction group decreased significantly (P<0.01). Conclusion Qingjin Huatan Decoction can reduce airway mucus hepersecrection of COPD by inhibiting ERK and p38 signal pathway on EGFR downstream.
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Objective To investigate the action mechanism of Qingjin Huatan Decoction on regulating COPD rats with mucus hypersecretion in airway. Methods Fifty 6-8 week old Wistar male rats were randomly divided into healthy group, model group, Qingjin Huatan Decoction group, clarithromycin group, 10 rats in each group. Except for healthy group, all the other groups were used the combination method of injecting LPS and smudging to establish COPD models. The last three groups were fed with normal saline, Qingjin Huatan Decoction, clarithromycin respectively for 30 days. On the 31st day of the experiment, the rats were put to death to take lung tissue. 6 rats from each group were chosen randomly, and the protein expressions of NE, EGFR and MUC5AC mRNA in lung tissue were detected by real-time fluorescent quantitative PCR. Results The expressions of NE, MUC5AC mRNA in lung tissue of COPD rats were higher than the healthy group. Compared with model group, the expressions of NE mRNA and MUC5AC mRNA in Qingjin Huatan Decoction group and clarithromycin group were markedly lower, while the expressions of NE, MUC5AC mRNA in Qingjin Huatan Decoction group were significantly lower than those in clarithromycin group. The expression of EGFR mRNA in Qingjin Huatan Decoction group was lower than that in model group. Conclusion Qingjin Huatan Decoction can inhibit mucus hypersecretion in airway through NE/EGFR/MUC5AC signal transduction pathway.
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Objective To observe the influence of Qizhi Yifei containing serum on the expression of MMP-9 and TIMP-1 mRNA in lung fibroblasts, and explore its mechanism. Methods Trypsin digestion method was used to extract fibroblasts from lung tissue in rats. All fibroblasts were cultured and trained to the fourth generation. Then they were randomly divided into control group, model group and drug serum group. The model group and drug serum group were firstly treated by DMEM with 0.002 5 μg/mL TGF-β1. The control group was treated by DMEM only. The control group and model group were then treated by DMEM with blank drug serum in concentration of 5%, and the drug serum group was treated by DEME with Qiahi Yifei containing serum in same concentration. After 48 and 72 hours, RT-PCR was performed to test the expression of MMP-9 mRNA and TIMP-1 mRNA of each group. Results After 48 hours, MMP-9 and TIMP-1 mRNA were significantly increased in model group and drug serum group compared with control group. There was no difference between model group and drug serum group. After 72 hours, MMP-9 mRNA was up-regulated in model group and was decreased in drug serum group compared with control group. There was no significant difference among the three groups on the expression of TIMP-1 mRNA. Conclusion Qizhi Yifei containing serum can decrease the up-regulated expression of MMP-9 mRNA in lung fibroblasts stimulated by TGF-β1.
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Objective:To preliminarily study the effect of Louqin Zhisou decoctions with the mechanism of clearing heat and resol-ving phlegm on the blood level of neutrophil elastase ( NE) in the rat model of acute exacerbation of chronic obstructive pulmonary dis-ease ( AECOPD) . Methods:The rat model of AECOPD was established by passive cigarette smoking, intratracheal instillation of li-popolysacchricle ( LPY) and intranasal instillation of staphylococcus aureus. Totally 60 AECOPD rats were randomly divided into 3 groups, the model control group (n=20), ambrohexel group (n=20), and Louqin Zhisou decoctions group (n=20). NE was detec-ted by ELISA. Results:Compared with that before the treatment, NE in ambrohexel group and Louqin Zhisou decoctions group was de-creased significantly(P<0. 01). Conclusion:Clearing heat and resolving phlegm method probably can decrease NE by reducing air-way mucus hypersecretion and obstruction in AECOPD rats.
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Objective To estimate TCM syndrome of rat model with COPD. Method The rat model with COPD was made by infusing the airway with endotoxin and fumed by cigarets. The macrography, pathological morphosis and lung function of the model were observed, and the features of their TCM syndrome were estimated. Result Model rats had these features:fur tarnished, yellow and unsmooth, blue nails, spirit depressed, activity decreased, less resistance and less bite, breathe deeply and increased after activity, lose weight campared with the control group, secretion increased in the airway. The volume of lung tissue increased with stagnated blood, the wall, smooth muscle and collogen of small airway thicken. Compared with the control group, the expiratory resistance (Re), maximal ventilatory volume (MVV), forced expiratory volume in 0.3 second/forced vital capacity (FEV0.3/FVC), forced expiratory flow between 25% and 35% of forced vital capatity (FEF25～75) of model all got degraded, the difference between the two groups was significant (P
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Objective To establish chronic obstructive pulmonary disease(COPD)rat models,and to study the intervention of the formula of nourishing Qi,activating blood circulation and dispersing phlegm recipe on the morphology.Method To establish rat COPD models by intratracheal instillation of lipopolysaccharide and exposure to cigarette smoke.To instill intervention drug daily either the formula of nourishing Qi,activating blood circulation and dispersing phlegm recipe or the roxithromythin,starting on the 20th,30th and 40th day of the experiment respectively(the groups was named h 20,h 30,h 40,r 20, r 30 and r 40 for short),and to observe the effect on the morphology by means of collagen staining and image analyzer.Results The pathological changes and lung function in the model group were accorded with the human COPD.In drug intervention groups,airway inflammation and epithelial proliferation were alleviated to different degree compared to the model group.In model group,the collagen deposition was increased predominant type I collagen compared to the health comtrol group,and the deposition in the drug intervention groups were decreased compared to the model group,according to the Sirius redpolarizing microscopy morphometry method.The thickness of the airway wall in the model group was significantly increased compared to the health control group(P