基于RhoA/ROCK1信号通路的舒肝和络醒脾方干预肝纤维化模型大鼠作用机制研究OA
Study on the mechanism of Shugan Heluo Xingpi Fang(舒肝和络醒脾方)intervention in hepatic fibrosis model rats based on the RhoA/ROCK1 signaling pathway
目的 基于Ras同源基因家族成员A(RhoA)/Rho相关卷曲螺旋蛋白激酶1(ROCK1)信号通路探讨舒肝和络醒脾方抗肝纤维化(HF)的作用机制.方法 将60只SPF级雄性Wistar大鼠随机分为空白组、模型组、阳性对照组及舒肝和络醒脾方高、中、低剂量组,每组10只.除空白组外,其余5组腹腔注射40％四氯化碳(CCl4)玉米油溶液构建HF大鼠模型,造模的同时,空白组予生理盐水灌胃,其余各组予相应药物灌胃,每日1次,连续干预8周.干预结束后,采用全自动生化分析仪检测各组大鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、白蛋白(ALB)、球蛋白(GLB)水平;采用苏木精-伊红(HE)染色、马松(Masson)染色观察肝脏组织病理形态变化;采用酶联免疫吸附法(ELISA)检测血清透明质酸(HA)、层粘连蛋白(LN)水平;采用实时荧光定量逆转录聚合酶链反应(RT-qPCR)、蛋白质免疫印迹法(Western blotting)分别检测肝脏组织中RhoA、ROCK1 mRNA及蛋白相对表达量.结果 与空白组比较,模型组大鼠血清ALT、AST、GLB、HA、LN水平及肝脏组织RhoA、ROCK1 mRNA及蛋白表达水平均明显升高,血清ALB水平明显降低(P＜0.05或P＜0.01);肝脏组织可见明显炎性细胞浸润、胶原纤维沉积及假小叶形成,胶原容积分数明显升高(P＜0.01).与模型组比较,阳性对照组及舒肝和络醒脾方高、中剂量组血清ALT、AST、GLB、HA、LN水平均明显降低,血清ALB水平均明显升高(P＜0.05或P＜0.01);舒肝和络醒脾方低剂量组血清ALT、AST、HA水平均明显降低(P＜0.05).与模型组比较,阳性对照组及舒肝和络醒脾方高、中、低剂量组大鼠肝脏组织炎性细胞浸润及纤维化程度均有所减轻,胶原容积分数及肝脏组织RhoA、ROCK1 mRNA及蛋白表达水平均明显降低(P＜0.05或P＜0.01).结论 舒肝和络醒脾方可能通过调控RhoA/ROCK1信号通路抑制肝星状细胞活化,进而延缓HF进展.
Objective To investigate the mechanism of Shugan Heluo Xingpi Fang(SHXF,舒肝和络醒脾方)against hepatic fibrosis(HF)based on the Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil containing protein kinase 1(ROCK1)signaling pathway.Methods Sixty SPF grade male Wistar rats were random-ly divided into a blank group,a model group,a positive control group,and high,medium,and low dose SHXF groups,with 10 rats in each group.Except for the blank group,the other five groups were intraperitoneally injected with a 40％carbon tetrachloride(CCl4)corn oil solution to establish the HF rat model.During modeling,the blank group received normal saline by gavage,while the other groups received the corresponding drugs by gavage once daily for 8 consecutive weeks.After intervention,serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),albumin(ALB),and globulin(GLB)were measured using an automated biochemical analyzer.Hematoxylineosin(HE)staining and Masson staining were used to observe histopathological changes in liver tissue.Serum levels of hyaluronic acid(HA)and laminin(LN)were detected by enzymelinked immunosorbent assay(ELISA).The relative mRNA and protein expression levels of RhoA and ROCK1 in liver tissue were measured by quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blotting,respectively.Results Compared with the blank group,the model group showed significantly increased serum levels of ALT,AST,GLB,HA,and LN,as well as significantly elevated mRNA and protein expression levels of RhoA and ROCK1 in liver tissue,while serum ALB level was significantly decreased(P＜0.05 or P＜0.01).Liver tissue exhibited obvious inflammatory cell infiltration,collagen fiber deposition,and pseudolobule formation,with a significantly in-creased collagen volume fraction(P＜0.01).Compared with the model group,the positive control group and the high and medium dose SHXF groups showed significantly decreased serum levels of ALT,AST,GLB,HA,and LN and significantly increased serum ALB levels(P＜0.05 or P＜0.01).The low dose SHXF group showed significantly re-duced serum levels of ALT,AST,and HA(P＜0.05).Compared with the model group,the positive control group and all SHXF dose groups exhibited alleviated inflammatory cell infiltration and fibrosis in liver tissue,along with signifi-cantly decreased collagen volume fraction and reduced mRNA and protein expression levels of RhoA and ROCK1 in liver tissue(P＜0.05 or P＜0.01).Conclusion Shugan Heluo Xingpi Fang may inhibit hepatic stellate cell activa-tion by regulating the RhoA/ROCK1 signaling pathway,thereby delaying the progression of hepatic fibrosis.
陈世玉;赵鲲鹏;张秋菊;王奎淞;李婕妠;郭嘉琪
甘肃中医药大学中医临床学院,甘肃 兰州 730101甘肃中医药大学中医临床学院,甘肃 兰州 730101甘肃中医药大学基础医学院,甘肃 兰州 730101甘肃中医药大学中医临床学院,甘肃 兰州 730101甘肃中医药大学中医临床学院,甘肃 兰州 730101甘肃中医药大学中医临床学院,甘肃 兰州 730101
医药卫生
肝纤维化舒肝和络醒脾方Ras同源基因家族成员A/Rho相关卷曲螺旋蛋白激酶1信号通路作用机制大鼠实验研究
hepatic fibrosisShugan Heluo Xingpi Fang(舒肝和络醒脾方)Ras homolog gene family member A/Rho-associated coiled-coil containing protein kinase 1 signaling pathwaymechanism of actionratsexperimental study
《甘肃中医药大学学报》 2026 (3)
1-8,8
甘肃省教育科技创新项目(2021CXZX-770).
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