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PTEN在CCl4诱导肝纤维化大鼠模型中的作用及益气活血方对其的影响
Role of phosphatase and tensin homolog deleted on chromosome ten in a rat model of carbon tetrachloride-induced liver fibrosis and the effect of qi-tonifying and blood-activating prescription
文章发布日期:2017年12月09日  来源:  作者:牛学敏,王宝玉,王洋,等  点击次数:454次  下载次数:96次

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【摘要】:目的 探讨第10号染色体缺失性磷酸酶张力蛋白同原物基因(PTEN)在CCl4诱导肝纤维化大鼠模型中的作用,阐明益气活血方调节PTEN阻止肝纤维化的分子机制。方法 选用27只Wistar雄性大鼠,随机分为3组,每组9只。其中肝纤维化组以CCl4诱导建立肝纤维化模型;益气活血方组在CCl4造模同时自拟以黄芪、丹参、云苓等中药为主的益气活血方进行干预实验;对照组以橄榄油腹腔注射。HE染色、Masson染色及胶原(Col1A1和Col4)免疫组化染色观察不同组别大鼠肝纤维化及胶原沉积程度;qRT-PCR、免疫组化及Western Blot检测TGFβ1、PTEN及其下游基因AKT、mTOR及p70S6K表达。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 肝纤维化组大鼠肝组织病理学可见窦周纤维化、汇管区纤维组织增生及胶原沉积、纤维间隔形成,伴随促肝纤维化基因TGFβ1 mRNA及蛋白表达上调,PTEN表达下调,而PTEN下游信号因子AKT、mTOR、p70S6K mRNA及磷酸化蛋白表达上调,与对照组比较差异均有统计学意义(P值均<0.01)。应用益气活血方组大鼠肝纤维化显著改善,PTEN表达较肝纤维化组明显增加(P<0.01),TGFβ1 mRNA及蛋白表达明显抑制(P值均<0.05),AKT、mTOR及p70S6K的mRNA及磷酸化蛋白表达较肝纤维化组明显减少(P值均<0.05)。结论 益气活血方可能通过调节PTEN及其下游信号因子的表达,阻止及逆转肝纤维化。
【Abstract】:Objective To investigate the role of phosphatase and tensin homology deleted on chromosome ten (PTEN) in a rat model of carbon tetrachloride (CCl4)-induced liver fibrosis and the molecular mechanism of action of qi-tonifying and blood-activating prescription in regulating PTEN and inhibiting liver fibrosis. Methods A total of 27 male Wistar rats were randomly divided into three groups, with 9 rats in each group. The rats in liver fibrosis group were treated with CCl4 to establish a model of liver fibrosis, and those in qi-tonifying and blood-activating prescription group were also treated with CCl4 to establish a model and then given a self-made qi-tonifying and blood-activating prescription containing Astragalus membranaceus, Salvia miltiorrhiza, and poria. The rats in the control group were given intraperitoneally injected olive oil. HE staining, Masson staining, and immunohistochemical staining of collagen type I alpha 1 (Col1A1) and collagen type Ⅳ (Col4) were performed to observe the degree of liver fibrosis and collagen deposition; qRT-PCR, immunohistochemistry, and Western blot were used to measure the expression of transforming growth factor-β1 (TGF-β1), PTEN, and downstream genes AKT, mTOR, and p70S6K. A one-way analysis of variance was used for comparison of continuous data between multiple groups and the least significant difference t-test was used for further comparison between any two groups. Results In the liver fibrosis group, liver pathology showed perisinusoidal fibrosis and fibrous tissue proliferation, collagen deposition, and formation of fibrous septum in the portal area; compared with the control group, the liver fibrosis group had significant increases in the mRNA and protein expression of TGF-β1, a significant reduction in the expression of PTEN, and significant increases in the mRNA and phosphorylated protein expression of AKT, mTOR, and p70S6K (all P<0.01). The qi-tonifying and blood-activating prescription group had a marked improvement in liver fibrosis; compared with the liver fibrosis group, the qi-tonifying and blood-activating prescription group had a significant increase in the expression of PTEN (P<0.01), significantly inhibited mRNA and protein expression of TGF-β1 (both P<0.05), and significant reductions in the mRNA and phosphorylated protein expression of AKT, mTOR, and p70S6K (all P<0.05). Conclusion Qi-tonifying and blood-activating prescription can prevent and reverse liver fibrosis, possibly by regulating the expression of PTEN and its downstream signal factors.
【关键字】:肝硬化;PTEN;益气活血;大鼠,Wistar
【Key words】:liver cirrhosis; phosphatase and tensin homology deteted on chromosome ten; REINFORCING QI ACTIVAT BLOOD; rats, Wistar
【引证本文】:NIU XM, WANG BY, WANG Y, et al. Role of phosphatase and tensin homolog deleted on chromosome ten in a rat model of carbon tetrachloride-induced liver fibrosis and the effect of qi-tonifying and blood-activating prescription[J]. J Clin Hepatol, 2018, 34(1): 122-128. (in Chinese)
牛学敏, 王宝玉, 王洋, 等. PTEN在CCl4诱导肝纤维化大鼠模型中的作用及益气活血方对其的影响[J]. 临床肝胆病杂志, 2018, 34(1): 122-128.

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