Effects Of Kangfuxin Solution Against Bleomycin-induced Pulmonary Fibrosis In Rats And Proliferation And Migration Of Mouse Fibroblasts

Objective 1.To evaluate the effect of Kangfuxin solution on anti-pulmonary fibrosis;2.To evaluate the effects of Kangfuxin solution on fibroblast proliferation and migration;3.To evaluate the effect of Kangfuxin solution on the cyclin Cyclin D1 and CKIS,and study the mechanism of Kangfuxin solution against pulmonary fibrosis,in order to provide laboratory evidence for the treatment of pulmonary fibrosis with Kangfuxin solution.Method SD rats were used as in vivo researchs,and the pulmonary fibrosis model was replicated by intratracheal instillation of bleomycin.After the Kangfuxin solution was administered,the general body signs such as body weight,lung wet weight and lung coefficient were observed.The characteristics of pathological of rat lung tissues were observed by H&E staining,Masson staining and Ashcroft scoring.The expressions of ?-SMA,MMP-1,MMP-3,MMP-9,TIMP-1,Collagen I and Collagen III in lung sections were detected by immunohistochemistry.Mouse primary lung fibroblasts were selected as in vitro researchs,and cells were purified by differential attachment method,and cells were identified by immunocytochemistry.The fibroblast viability was detected by CCK-8 test.The effect of Kangfuxin solution on fibroblast proliferation was detected by Cell counting method.The expression of Cyclin D1,p15ink4 b,p18ink4c and p19 arf genes in fibroblasts was detected by RT-PCR.The effect of Kangfuxin solution on the migration of fibroblasts were investigated by Transwell method.Result 1.Evaluation of Kangfuxin solution's anti-pulmonary fibrosis: General physiques: Kangfuxin solution and dexamethasone decresed the lung wet weight and lung coefficient decreased significantly but increased body weight of pulmonary fibrosis rats which induced by BLM.2.Lung histopathology: H&E staining showed that the lung tissue of the model group was severely damaged,and a large number of alveolar collapse formed fibrotic scar with diffuse collagen fibrosis,the alveolar wall was thickened,the number of alveoli was reduced.The Kangfuxin group and the dexamethasone group improved the above lesions.The improved Ashcroft scores showed that the Ashcroft scores of the Kangfuxin solution treatment group and the dexamethasone group were significantly lower than those of the bleomycin group.Masson staining showed that the collagen level of the rats in the Kangfuxin group and the dexamethasone group was significantly reduced.3.Pulmonary fibrosis lung tissue related biological indicators: Immunohistochemistry results showed that compared with the model control group,the expression of lung tissue ?-SMA,MMP1,MMP3,MMP9,TIMP-1,Collagen I and Collagen III proteins in the treatment group were significantly reduced.Conclusion 1.Kangfuxin solution can improve the general signs and lung tissue damage of rats with pulmonary fibrosis induced by BLM,and play a protective role in pulmonary fibrosis;2.Kangfuxin solution can reduce collagen deposition and alveolar structure destruction in rats with pulmonary fibrosis,and supress the content of biological markers of pulmonary fibrosis of MMP1,MMP3,MMP9,TIMP-1,?-SMA,Collagen I and Collagen III.3.Kangfuxin solution can suppress the proliferation of fibroblasts in vitro and in vivo.4.Kangfuxin solution can inhibit fibroblast migration in vitro.5.Kangfuxin solution may exert anti pulmonary fibrosis effects by inhibiting the expression of cyclin Cyclin D1 and promoting the expression of CKIS to inhibit fibroblast proliferation.