Slit2/Robo4/Rho信号通路在野百合碱诱导的大鼠肝窦阻塞综合征中的变化及意义  

Slit2/Robo4/Rho signaling pathway in monocrotaline-induced hepatic sinusoidal obstruction syndrome in rat

作　　者：徐魁 任晓非[1] 宋育林[1] Xu Kui;Ren Xiaofei;Song Yulin(Dept of Gastroenterology,The First Affiliated Hospital of Anhui Medical University,Anhui Key Laboratory of Digestive Diseases,Hefei 230022)

机构地区：[1]安徽医科大学第一附属医院消化科,安徽省消化系统疾病重点实验室,合肥230022

出　　处：《安徽医科大学学报》2021年第11期1756-1761,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽高校自然科学研究项目(编号:KJ2016A337);2016-2018年安徽高校科研平台创新团队建设项目(编号:皖教秘科[2015]49号)。

摘　　要：目的探讨Slit2/Robo4/Rho信号通路在野百合碱(MCT)导致肝窦阻塞综合征(HSOS)发病中的作用。方法34只SD大鼠随机分为正常对照组和实验组。实验组大鼠给予MCT灌胃(160 mg/kg),分批在造模后第1、2、4天随机处死,正常组第4天处死。收集血液样本和肝组织,测定血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)水平,观察肝脏组织学变化,测定肝脏α-平滑肌肌动蛋白(α-SMA)表达和肝脏Slit2、Robo4、RhoA、Rac1、Cdc42及基质金属蛋白酶(MMP)-2、MMP-9 mRNA和蛋白的变化。结果大鼠单次灌胃MCT(160 mg/kg)后,第1天肝窦淤血、肝细胞变性和中央静脉内皮损伤;第2天肝窦淤血扩张明显;第4天肝脏病变加重,可见窦周纤维化。与正常对照组比较,各实验组大鼠的肝指数均升高(P <0.05),实验组第2、4天AST水平升高(P <0.01),第2天ALT升高(P <0.01)。与正常对照比较,各实验组α-SMA表达升高(P <0.01)。各实验组Slit2和Robo4 mRNA和蛋白低于正常对照(P <0.01),随着给药时间的延长,实验组大鼠Slit2和Robo4 mRNA和蛋白的表达逐渐降低(P <0.05);而各实验组RhoA、Rac1、Cdc42及MMP-2、MMP-9 mRNA和蛋白高于正常对照组,且表达逐步升高(P <0.05)。结论单次MCT摄入可使大鼠发生HSOS,且肝损伤呈进行性加重。Slit2/Robo4/Rho信号通路、肝星状细胞激活及MMP-2、MMP-9升高可能是MCT致HSOS发生的重要作用机制之一。Objective To explore the function of Slit2/Robo4/Rho signaling pathway in monocrotaline(MCT)-induced hepatic sinusoidal obstruction syndrome(HSOS).Methods Thirty-four rats were randomly divided into control group and experimental group.The rats in experimental group was established with MCT by gastric lavage(160 mg/kg),and randomly sacrificed after model making for 1 day,2 days and 4 days.Serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured by chemical method.Liver tissue damage was observed by HE staining and Masson staining.α-smooth muscle actin(α-SMA)protein expression in rat liver was measured by immunohistochemistry.The related indexes of Slit2/Robo4/Rho signaling pathway were measured by Western blot and RT-qPCR.Results After a single intragastric administration of MCT(160 mg/kg),the congestion of hepatic sinusoid,the degeneration of hepatocyte and central venous endothelial cell injury of the 1-day group were observed,the congestion and dilatation of hepatic sinusoid of the 2-day group was obvious,hepatic pathological changes were more serious of the 4-day group and perisinusoidal fibrosis was seen in the 4-day group.Compared with the control group,the liver index of the experimental group increased(P<0.05)serum AST of the 2-day group and the 4-day group increased(P<0.01),serum ALT of the 2-day group increased(P<0.01).α-SMA protein expression in rat liver of all treatment group was significantly higher than the control group(P<0.01).The expressions of Slit2,Robo4 mRNA and protein in each experimental group was lower than those of the control group(P<0.01)and gradually decreased with the prolonged administration time(P<0.05),while the expressions of RhoA,Rac1,Cdc42,matrix metalloproteinase(MMP)-2,MMP-9 mRNA and protein in each experimental group were higher than those of the control group(P<0.01)and gradually increased(P<0.05).Conclusion Rat HSOS was induced by single MCT,and rat liver injury was progressively aggravated.Slit2/Robo4/Rho signaling pathway,hepatic stellate ce

关 键 词：野百合碱 肝损伤 Slit2/Robo4 RHO 肝窦阻塞综合征 

分 类 号：R575[医药卫生—消化系统] R99[医药卫生—内科学]