Research On Function And Mechanism Of Preventing And Treating Hepatic Fibrosis In Rats Through The Methods Of Expelling Wind And Activating Blood Circulation

Objective:To discuss the mechanism of action of expelling wind and activating blood circulation on resisting hepatic fibrosis; to provide theoretical basis for expelling wind and activating blood circulation applied to the clinical treatment of blood stasis; to broaden in therapy pathway of blood stasis.Method:With the use of experimental methods, using carbon tetrachloride-induced liver fibrosis in a rat model as study, observing the effect of the methods of expelling wind and activating blood circulation on the aspects of liver tissue, serum biochemical, gene pathway.95male SD rats were classified into groups6:normal contorl, pathological model, colchicine group, xia yu xue tang group(dahuang10g,taoren10g,tubiechong6g), the methods of expelling wind and activating blood circulation] group(fangfeng12g, baizhi12g, qianghuo12g), the methods of expelling wind and activating blood circulation2group(fangfeng12g, baizhi12g, qianghuo12g, dahuang10g,taoren10g,tubiechong6g). Besides the normal group, others were made into HF models by CC14subcutaneous injection. Meanwhile, feed each group same amount of normal saline by gavage after model checking succeeded the eighth weeks later. After12weeks later,sacrified the animals, remove spare blood and liver tissue. Testing and doing statistical analysis the serum concentration of HPC-Ⅲ、ColⅣ-C、FN、HA of each group. Liver tissues were embedded in paraffin, sliced made after HE staining, and observed degeneration of liver cells and the situation of fibrous tissue hyperplasia of each group in the light microscope. After uranyl acetate and lead citrate double staining, observed the changes in liver cell ultrastructure in the transmission electron microscope. Using liquid nitrogen flash freezer grinding homogenate method to treat fresh liver issue, total RNA was extracted with Trizol, and using gene chips to detect the gene expression changes, obtaining differentially expressed gene profile between each group, analyzing the impact of differences and similarities of each treatment method in related signaling pathway genes. Liver cell edema, changes in fat eased in the methods of expelling wind and activating blood circulation group1, the methods of expelling wind and activating blood circulation group2, colchicine group and xia yu xue tang group, liver connective tissue proliferation better than the pathological model, all can improve the phenomenon of sinusoidal endothelial cell apertures dicreased or disappeared, improve organelle structure, reduce the deposition of collagen fibers, the denseness of HPC-Ⅲ, FN in serum were decreased, and the methods of expelling wind and activating blood circulation group1lower the denseness of HPC-Ⅲ in serum of hepatic fibrosis in rats better than other groups. Comparison of differentially in genes pathway, the methods of expelling wind and activating blood circulation group1exerts pharmacological effects by the means of immune response, inflammation, fat metabolism, cytoskeleton and other related genes and pathways; genes after treatment are closed to its in colchicine group and xia yu xue tang group, description wind-expelling herbs can resist fibrotic of experimental rats. Compare with the other groups, the methods of expelling wind and activating blood circulation group2has no significant differences.Conclusion:Consist separate wind drugs of the methods of expelling wind and activating blood circulation group1could improve the degree in carbon tetrachloride-induced liver fibrosis in rats with hepatic fibrosis, can prevent liver fibrosis in rats, mechanism may be same to expelling wind and activating blood circulation group1, it could lower the denseness of HPC-Ⅲ, improve the degree, improve pathological structure of liver cells, reduce the proliferation of fibrous tissue; changes related to immune response, inflammtion, fat metabolism, cytoskeleton and other related genes and pathways, demonstrate it exerts anti-fibrotic effect from multipath, multilevel and multiple target points.Compound wind-expelling herbs and activating blood circulation in to the methods of expelling wind and activating blood circulation group2, its have effect on the liver cell edema, changes in fat eased, liver tissue hyperplasia and the dense of HPC--Ⅲ, improve liver fribs, reduce the process to prevent liver fibrosis in rats. However, the difference was not significantly different comparing gene pathway, the effect is not significant. Promote the choice of wind-expelling herbs and traditional herbs, In strict accordance with the prescription compatibility, shall be based on symptom, drug cold and heat.