| This thesis has altogether three parts.Part one:literature review The main tissue cells and inflammatory cells involved in the pathogenesis of COPD and the important mechanisms was reviewed. The pathological changes of small airway remodeling and there evaluation methord also summarized here. In Chinese medicine literature review, we first summarized the etiology and pathogenesis, symptom research and clinic outcome research. The main methords we used in the research of Chinese on COPD including the correlation research between etiology pathogenesis and many features of COPD, and the molecular biology index changes were all summarized here.Part two:Theoretical research There is surely a necessity to identify the etiology and pathogenesis of Chinese medicine from long time smoking exposure to COPD. According to the content from ancient Chinese medicine book, we summarized the nature of tobacco first. With the knowledge we hanv on the nature of lung, we attempted to link this to COPD.Part three:ResearchObjective To identify the mechanism of QiZhi YiFei granules in treating small airway of COPD by regulating ECM, and its concrete effective targets.Methods 72 Wista rats were randomly divided into control group, model group and treatment group. Cigarette smoking exposure and intratracheal LPS instillation were combined together to mode COPD. A homemade box was used to accomplish this process.8 cigarettes were lighted each time with 10 rats in box for 30 minutes.7 days per week,30 minutes per day. LPS installation were done in day 1, day 11 and day 21. All this smoking exposure lasted for 28 days, excepted the days in LPS installation. QiZhi YiFei granules were given intragastrically once a day from day 21, rats in treatment group were given saline in the same dose and same manner. Rats in control group had no treatment. All rats were killed on the 30th day,40th day, and 60th day in batches. The histology and collagen fibre were test by Hematoxylin-Eosin staining and Van Gieson staining. The expressions of TGF-(3i, Smad3, Smad4, Smad7, MMP-9, TIMP-1, COL ?,COL ? proteins around small airway were tested by using immunohistochemistry staining. The expression of TGF-?1mRNA,Smad3mRNA, Smad4mRNA, Smad7mRNA,MMP-9mRNA, TIMP-1 mRNA were tested by using real-time fluorescence quantitative polymerase chain reaction(RT-PCR). The expressions of proteins and the expressions of the genes were compared to indentify weather there was some relationship between each other.Results1:HE and VG stainingSmall airway wall thickness in model group was increased compared to control group at each stage, and it was decreased in treatment group in day 40 when compared to model group; Content of collagen in model group was increased in model group compared to control group at each stage, and was decreased in treatment at each time compared to control group, all changes above were all have a significant difference.2:Immunohistochemistry stainingThe expression of TGF-?1 in model group was increased compared to control group at each stage, and was decreased in treatment at each time compared to control group. Smad3 in model group was increased compared to control group in 30th day and 60th day, and was decreased in treatment at each time compared to control group; Smad4 was increased compared to control group in 30th day, and was decreased in treatment in 30th day and 40th day compared to control group; Smad7 in model group was decreased in 30th day and was increased in 60th day, the protein in treatment group was all increased compared to model group in each time. There was no difference of the expression of MMP-9 in treatment group compared to model group at each stage. TIMP-1 in treatment group in 30th day was decreased compared to model group, all changes above were all have a significant difference. The ratio of MMP-9/TIMP-1 was significantly decreased in 30th day and 40th day in model group compared to control group, and was significantly increased in treatment.COL I protein in model group was decreased in 30th and 60th day compared to control group, the protein in treatment group was decreased in 40th day and increased in 60th day; COL III protein in model group was increased in each stage compared to control group, and was decreased in 30th day and 60th day.3:RT-PCRThe expression of protein TGF-?1mRNA in model group was increased in 40th day and 60th day compared to control group, and was decreased in treatment at each time compared to control group. Smad3mRNA in model group was decreased compared to control group in 30th day, and was then increased in treatment group; Smad4mRNA was decreased in 30th day and increased in 60th day compared to control group, and then was increased in treatment in 60th day compared to control group; Smad7mRNA in treatment group was significantly decreased in 40th day and 60th day compared to model group. all changes above were all have a significant difference. There was no difference of the expression of MMP-9mRNA and TIMP-1 mRNA between each group and each time. The expressions of each proteins around small airways and the gene expressions we examined of whole tissue of airways and pulmonary parenchyma have no relationship.Conclusion QiZhi YiFei granules can effectively intervene the development of small airway remodeling of COPD by reducing the deposition of ECM around airway. It can not only down-regulate the TGF-?1/Smads signal pathway to inhibit the synthesis of ECM, but also increased the ratio of MMP-9/TIMP-1 to promote the its degradation. The concrete targets of it is to decrease the expression of TGF-?1, Smad3, Smad4 and to increase Smad7 protein. The mechanism in which QiZhi YiFei granules can increase the ratio of MMPs/TIMPs is unclear after this reach. The expressions of each proteins around small airways and the gene expressions we examined of whole tissue of airways and pulmonary parenchyma have no relationship after research, this result suggest us that the pathogenesis of small airway remodeling may differ from emphysema in COPD. |
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