壮肝逐瘀煎对肝纤维化大鼠肝脏PI3K、Akt表达的影响

目的:观察壮肝逐瘀煎对肝纤维化大鼠肝脏PI3K/Akt信号传导通路的影响,探讨其抗纤维化的作用机制。方法:随机选取14只大鼠作为正常对照组,剩余24只大鼠采用CCL4复合因素大鼠肝纤维化模型造模方法[3]进行肝纤维化造模,于第4周末随机处死正常对照组及造模大鼠各4只,做病理检测证实造模组肝纤维化形成与否。其余造模大鼠随机分为病理模型组、壮肝逐瘀煎治疗组,每组10只。病理模型组、壮肝逐瘀煎治疗组于造模结束后即分别给药,每组给药6周。用药后处死所有大鼠,获取肝组织,HE染色光镜观察肝组织结构变化,免疫组化染色分析PI3K、Akt表达。结果:光镜观察,正常对照组肝小叶完整,结构清晰,细胞以静脉为中心呈条索状向四周放射状排列。病理模型组肝小叶结构紊乱,肝细胞水肿明显,较多脂肪变性,部分有坏死,纤维隔内炎性细胞浸润,较多粗大胶原纤维分割、包绕肝小叶。壮肝逐瘀煎治疗组与病理模型组比较肝小叶结构较清楚,肝细胞水肿及变性不明显,炎细胞浸润较少,胶原纤维增生较少,纤维疏松变窄。肝组织PI3K、Akt以胞浆内出现棕黄色颗粒或团块判断为阳性细胞。病理模型组、壮肝逐瘀煎治疗组PI3K、Akt的表达值较正常对照组增高,比较有统计学意义(P0.0.1);壮肝逐瘀煎治疗组肝组织PI3K、Akt的表达值较病理模型组下降,比较有统计学意义(P0.01)。结论:壮肝逐瘀煎能调控HSC中凋亡因子PI3K、Akt蛋白的表达,明显改善HF大鼠肝脏的病理变化,具有明显的抗HF作用。

Influence of Liver-Reinforcing and Blood-Stasis-Expeling Decoction on the Expression of PI3K and Akt in the Hepatic Fbrotic Rats

Objective To observe the influence of Liver-Reinforcing and Blood-Stasis-Expeling Decoction on the expression of Pl3 K and Akt in the hepatic fbrotic rats,and investigate the anti-fibrosis mechanism.Methods:Fourteen rats randomly selected were normal control group while the other 24 rats were establish into hepatic fbrotic models with CC14 compound factors.After 4 weeks,4 rats in normal control group and 4model rats were executed and conducted pathological test to confirm the formation of hepatic fbrotic models.The rest of the model rats were randomized into pathological model group,Liver-Reinforcing and Blood-Stasis-Expeling Decoction group,10 rats in each group.After the molding,both groups were given administration for 6 weeks and after that all rats in both groups were executed with drugs.The livers of the rats were obtained.The structural changes of liver tissue were observed by HE staining and the expression of PI3K and Akt was determined by immunohistochemical staining.Results:With light microscopy,the structure of hepatic lobules in normal control group was clear,and the cells centered on the veins and radially arranged peripherily.The structure of hepatic lobules in pathological model group was disordered.,the fibrous spacing was significantly thinner and the expression of PI3K and Akt in liver tissue was significantly decreased in the treatment group (P ＜0.01).The swollen of hepatic cells was obvious and much fat degenerated and some even became necrosis.Inflammatory cell infiltrated in the fibrotic septum.Many thick collagenous fibers split and enveloped around the hepatic lobules.Compared with model group,the structure of hepatic lobules in Liver-Reinforcing and Blood-Stasis-Expeling Decoction group was clear;the swelling and degeneration of hepatic cells were not obvious;Few inflammatory cell infiltrated,the collagenous fibrogenesis was not obvious and fibers became loose and narrow.The PI3K and Akt in the liver tissue were determined as positive cells by the brown granules or masses appeared in the intracytoplasm The expression values of pathological model group and Liver-Reinforcing and Blood-Stasis-Expeling Decoction group were higher than that in normal control group,with statistical significance (P ＜ 0.01);but the value in Liver-Reinforcing and Blood-Stasis-Expeling Decoction group decreased compared with mode] group,with statistical significance (P ＜0.01).Conclusion:Liver-Reinforcing and Blood-Stasis-Expeling Decoction can obviously improve the liver pathological changes of HF rats,and has obvious anti-HF effects.The mechanism may correlate with its regulation of the protein expression of the apoptosis factors of PI3K and Akt in the HSC.