Effect Of Jianpiruanganfang On ?-SMA And M-RNA Expression In Rats With Hepatic Fibrosis Induced By Carbon Tetrachloride

ObjectiveTo study the effect of Jianpiruanganfang on oa-smooth muscle actin (a-SMA) and m-RNA expression in rats with hepatic fibrosis induced by carbon tetrachloride(CCl4), and to explore the role of Jianpiruanganfang prevention and controlling of the hepatic fibrosis and its mechanism, providing experimental basis for clinic treatment of hepatic fibrosis in traditional Chinese medicine.MethodsRat models of hepatic fibrosis were made by intraperitoneal injection of 30%CCla and 70% olive oil mixed solution with 0.2mL/100g, two times a week for a total of 4 weeks. The expression of a-SMA in rats with hepatic fibrosis was detected by ABC immunohistochemical method and the m-RNA expression was determined by Real-time PCR method. The H-E staining and Mallory staining were performed to observe the pathological changes of hepatic fibrosis in rats.Results1. Effects of Jianpiruanganfang on the general state and body weight change The rats in model group appeared to be dark yellow. fur lackluster, sluggishness,redness and swelling of anal region with a weight gain of 50 -80g, and the mortality rate was 16.7% after four weeks of feeding. Compared with the model group. there was an increase of food and water intake in three Jianpiruanganfang groups (high.medium and low doses), and the rats had a shiny fur, loose stools sometimes while general situation was better. At the end of four weeks, the mortality rate of the high and the low dose groups was 8.3%,and the average weight of the three groups was increased by 80 -100g.2. Liver morphology and liver wet weight in rats(1) Rats liver morphologyCompared with the normal group, the liver color of the model group rats was light and dark red. The liver volume was larger than the normal group with nodular surface, hard texture and the edges were obtuse and round with connective tissue adhesions. The liver of Fuzhenghuayu capsule group and Jianpiruanganfang high,medium and low dose group rats became darker with smooth surface, slight surface nodules and hard texture, and liver size slightly increased compared with the normal group, but the overall situation was better than the model group.(2) Liver wet weightCompared with the normal group, the liver weight of model group rats increased,the difference was statistically significant (P<0.05). Compared with the model group,the liver weight of the rats in each group significantly decreased (P<0.05), but the decreases in liver weight among the high, medium and low dose groups do not show certain drug dependence.3. Histopathology observation of liver in rats(1) H-E stainingIn the model group rats, the liver cells exhibited swelling and deformation with the false liver acinus formed and hepatic cord arranged in disorder, and the local liver cells underwent fatty degeneration with the collagen fiber tissue area expanded,especially severe in the hepatic periportal. Compared with the model group, in all dose groups of Jianpiruanganfang and Fuzhenghuayu capsule group, the lesions of rat liver cells were mild with hepatic cord arranged in disorder and the false liver acinus formed, and fatty lesions were mild with formation of a small amount of fat-like vacuoles and the decrease of collagen fibers tissue area.(2) Mallory stainingThe liver cells of model group rats were denatured with the formation of false liver acinus and a large number of fat vacuoles and massive deposition of fiber connective tissue, and the structures of liver lobules and periportal area are disrupted.In all dose groups of Jianpiruanganfang and Fuzhenghuayu capsule group,false liver acinus formed with deposition of a little fibrous connective tissue in the liver perportal area each group and collection of a small amount of fat vacuoles. The structure of liver lobules was damaged in a certain extent, but the overall extent of damage was less slight compared with the model group.4. Grading of hepatic fibrosisIn the model group, the false liver lobules were formed in most of the rat liver tissue in the S3 stage. In all dose groups of Jianpiruanganfang and Fuzhenghuayu capsule group, a few collagen fibers accumulated in liver but most of them are in the S2 stage. The hepatic fibrosis was relieved each treatment group.5. Expression of a-SMA in rat liverCompared with the normal group, the expression of a-SMA in the model group increased significantly (P<0.05). There was a significant difference between Fuzhenghuayu capsule group and the model group (P<0.05). The difference was statistically significant on the expression of a-SMA between the three dose groups of Jianpiruanganfang and the model group (P<0.05) and the expression of a-SMA increased gradually among three dose groups, indicating certain drug dependence.6. The m-RNA expression of a-SMA in rat liverCompared with the normal group, the m-RNA expression of a-SMA in the model group increased significantly (P<0.05). Compared with the model group, there was a significant difference in Fuzhenghuayu capsule group (P<0.05) and the difference was statistically significant on the m-RNA expression of a-SMA in the high, medium and low dose groups of Jianpiruanganfang (P<0.05). The m-RNA expression of a-SMA increased gradually among the high, medium and low dose groups, showing certain drug dependence.ConclusionThe mechanism of the Jianpiruanganfang for anti-hepatic fibrosis effect may be related to the inhibition of a-SMA and m-RNA expression in rat liver.