基于RhoA/ROCK通路探讨软肝冲剂对CCl4致大鼠肝纤维化的作用机制  

Exploring the Mechanism of Ruangan Granule on CCl4-induced Hepatic Fibrosis in Rats Based on RhoA/ROCK Pathway

作　　者：李楠楠[1] 孟宪生[1] 郭文昭 薛立平[2] 卢秉久[2] LI Nannan;MENG Xiansheng;GUO Wenzhao;XUE Liping;LU Bingjiu(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;The Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China)

机构地区：[1]辽宁中医药大学,沈阳110847 [2]辽宁中医药大学附属医院,沈阳110032

出　　处：《中国现代应用药学》2024年第16期2192-2198,共7页Chinese Journal of Modern Applied Pharmacy

基　　金：辽宁省重点研发计划项目(2020JH2/10300066);辽宁省自然科学基金计划项目(2022-BS-200)。

摘　　要：目的基于RhoA/ROCK通路探究软肝冲剂对CCl_(4)致大鼠肝纤维化的保护作用及其分子机制。方法大鼠灌胃给予2 mL·kg^(−1)40%CCl_(4)(CCl_(4)∶橄榄油=2∶3)每周2次,共8周,建立大鼠肝纤维化模型。造模第6周给予软肝冲剂干预8周,采用Masson染色、脏器指数、生化指标评价软肝冲剂对CCl_(4)致大鼠肝纤维化的作用;采用RT-PCR、Western blotting及免疫荧光组织化学法检测大鼠肝脏组织中CollⅠ、α-SMA、RhoA、ROCK 1 mRNA和蛋白质的表达。结果脏器指数和生化指标结果显示,与模型组比较,软肝冲剂中剂量组能降低大鼠的肝指数(P<0.01);软肝冲剂高、中剂量组能降低大鼠的脾指数(P<0.01或P<0.05);软肝冲剂高、中、低剂量组均能提高大鼠的胸腺指数(P<0.05或P<0.01);软肝冲剂可以不同程度降低大鼠的血清中AFP、ALT、AST水平(P<0.05或P<0.01)。Masson染色结果显示,软肝冲剂可以显著降低大鼠肝脏胶原纤维增生。RT-PCR、免疫荧光组织化学结果显示,软肝冲剂可以显著降低大鼠肝脏组织中CollⅠ、α-SMA、RhoA、ROCK 1 mRNA和蛋白质的表达。Western blotting结果显示,与模型组比较,软肝冲剂各剂量组均能降低CollⅠ、α-SMA、ROCK 1蛋白质的表达,软肝冲剂中、低剂量组均降低RhoA蛋白质的表达。结论软肝冲剂对CCl_(4)致大鼠肝纤维化的治疗作用可能与抑制RhoA/ROCK通路有关。OBJECTIVE To explore the mechanism of Ruangan Granule against CCl_(4)-induced hepatic fibrosis in rats based on RhoA/ROCK pathway.METHODS The rat hepatic fibrosis model was established by intragastric administration of 40%CCl_(4)(2 mL·kg^(−1))(CCl_(4)∶olive oil=2∶3),twice a week for 8 weeks.After the intervention of Ruangan Granule at 6 week for 8 weeks,Masson staining,organ index and biochemical indexes were used to evaluate the effect of Ruangan Granule on CCl_(4)-induced hepatic fibrosis in rats.RT-PCR,Western blotting and immunofluorescent tissue chemistry was used to detected the expression of CollⅠ,α-SMA,RhoA,ROCK 1 mRNA and protein in rats.RESULTS The organ index and biochemical analysis indicated that,compared with the model group,the liver index of rats was decreased in the middle dose group(P<0.01),and the spleen index was decreased in the high and middle dose groups(P<0.05 or P<0.01);high,middle and low dose of Ruangan Granule could increase the thymus index of rats(P<0.05 or P<0.01);Ruangan Granule could reduce the levels of AFP,ALT and AST in the serum(P<0.05 or P<0.01).The results of Masson staining showed that Ruangan Granule could significantly reduce the proliferation of collagen fibers in rat liver.RT-PCR,immune fluorescent tissue chemistry results showed that Ruangan Granule could significantly downregulate the expression of CollⅠ,α-SMA,RhoA,and ROCK 1 mRNA and protein.Western blotting results showed the protein expression of Colli,α-SMA and Rock-1 were decreased in all groups.The protein expression of RhoA was decreased in middle and low dose groups of Ruangan Granule.CONCLUSION The therapeutic effect of Ruangan Granule on CCl_(4)-induced hepatic fibrosis in rats may be related to the inhibition of RhoA/ROCK pathway.

关 键 词：软肝冲剂 肝纤维化 RhoA/ROCK通路 作用机制 

分 类 号：R966[医药卫生—药理学]