健脾软肝方预防给药降低TGF-β1表达延缓肝纤维化进程研究

Research of Jianpi Ruangan Decoction Preventive Administration on Delaying Process of Hepatic Fibrosis by Reducing the Expression of TGF-β1

目的：研究自拟健脾软肝方预防给药对CCl4肝纤维化大鼠的作用,着重分析其对转化生长因子-β1（Transforming Growth Factor-β1,TGF-β1）的影响。方法：60只清洁级雄性Wistar大鼠按照随机数字表法分为正常组,CCl4模型组,健脾软肝方低、中、高剂量组以及阳性药物扶正化瘀胶囊组,n=10。适应性喂养1周后给予CCl4腹腔注射复制肝纤维化大鼠模型,期间所有大鼠均接受预防给药,其中正常组和CCl4模型组给予生理盐水灌胃（1 m L·kg^-1·d^-1）,阳性药物组给予扶正化瘀胶囊（0.405 g·kg^-1·d^-1）灌胃,健脾软肝方3个剂量组给予健脾软肝冲剂（2.7 g·kg^-1·d^-1）、（5.4 g·kg^-1·d^-1）和（10.8 g·kg^-1·d^-1）灌胃[1次/（kg·d）],全程密切监测大鼠一般情况,4周后处死大鼠检测肝功能、肝纤维化指标、肝脏病理学,同时Western blot检测TGF-β1的表达特点。结果：与正常组相比,CCl4模型组毛色变暗,食欲减退,3个预防给药组和扶正化瘀胶囊组上述症状明显好转,同时健脾软肝方高剂量组明显好于扶正化瘀胶囊组;与正常组相比,模型组ALT和AST以及肝纤维化指标HA、PCIII、IV-C和LN明显增加（P〈0.05）,3个预防给药组和扶正化瘀胶囊组则可以明显降低其含量（P〈0.05）;HE及Masson染色可见,与正常组相比,CCl4模型组肝组织切片肝细胞肿胀明显,肝窦以及门静脉周围结缔组织明显增生,有假小叶形成,3个预防给药组和扶正化瘀胶囊组上述症状明显好转;与正常组相比,模型组TGF-β1表达明显升高（P〈0.05）,健脾软肝方3个预防给药组则可以明显降低其表达（P〈0.05）。结论：健脾软肝方预防给药可以明显延缓CCl4肝纤维化大鼠进展,其作用机制可能是通过降低TGF-β1表达而实现。

Objective：To investigate whether the Jianpi Ruangan Decoction has the affection of delaying the development of CCl4-induced hepatic fibrosis in rats or not,especially its effect on the transforming growth factor-β1（TGF-β1）. Methods：60 male Wistar rats were randomly divided into the normal group,CCl4 model group,low,middle,high dose of Jianpi Ruangan Decoction treatment group and Fuzheng Huayu Capsule intervention group（n=10）. All of rats were received the adaptive feeding one week,after CCl4 was injected intraperitoneally into them to build the hepatic fibrosis model. Then the rats in normal group and the CCl4 model group were given normal saline treatment [1 m L/（kg·d）],the rats in the positive drug group was given Fuzheng Huayu Capsule[0.405 g/（kg·d）],and the rats in the low,middle,high Jianpi Ruangan Decoction groups were given 2.7,5.4 and 10.8 g/（kg·d）Jianpi Ruangan Decoction. After 4 weeks,the weight of liver and body were measured,and the liver index was calculated,next detected the liver function,the methods of HE and collagen staining were used to observe the liver pathology,at last the expression characteristics of TGF-β1 was detected by Western blotting. Results：Compared with the normal group,the rat in CCl4 model group had some hairs darker and loss of appetite（P〈0.05）. Those symptoms in Jianpi Ruangan Decoction and Fuzheng Huayu Capsule groups were significantly improved,while we also could get the high-dose of Jianpi Ruangan Decoction group was significantly better than the Fuzheng Huayu Capsule group for those symptoms. Compared with the normal group,ALT,AST,HA,PCIII,IV-C and LN in the model group were significantly increased（P〈0.05）,those indexs in the Jianpi Ruangan Decoction and Fuzheng Huayu Capsule groups were significantly lowed（P〈0.05）. Compared with the normal group,the hepatocytes in the CCl4 model group showed an obvious swelling and some fat particles were appear in the hepatic sinusoids and connective tissue,the Jianpi Ruangan Decoction and Fuzheng Huayu Capsule groups showed obvious improvement. Compared with the normal group,the expression of TGF-β in the model group was significantly increased（P〈0.05）,the Jianpi Ruangan Decoction groups could significantly reduce the expression of TGF-β1（P〈0.05）. Conclusion：Jianpi Ruangan Decoction can delay the progression of CCl4-induced hepatic fibrosis. The mechanism may be related with it can reduced the expression of TGF-β.