芪甲柔肝方及其拆方调控VEGF/SRF/c-FOS通路与改善肝纤维化大鼠肝窦毛细血管化的机制研究  

Mechanism of Qijia Rougan Decoction and its disassembled formulas on regulation of VEGF/SRF/c-FOS pathway and improvement of hepatic sinusoidal capillaryization in rats with hepatic fibrosis

作　　者：刘进[1] 许欣怡 刘悸斌 冯全生[1] 苏悦[1] LIU Jin;XU Xin-yi;LIU Ji-bin;FENG Quan-sheng;SU Yue(Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China)

机构地区：[1]成都中医药大学,四川成都610075

出　　处：《中国中药杂志》2024年第20期5528-5538,共11页China Journal of Chinese Materia Medica

基　　金：四川省科技厅项目(2022YFS0409);成都中医药大学杏林学者项目(BSH2020014);成都中医药大学校院联合创新基金项目(LHJJ202402016)。

摘　　要：基于血管内皮生长因子(VEGF)/血清应答因子(SRF)/原癌基因c-FOS(c-FOS)通路及肝窦毛细血管化揭示芪甲柔肝方及其拆方抗肝纤维化的机制。将大鼠随机分为空白组(6只)和造模组(28只),造模组大鼠采用皮下注射40%四氯化碳(CCl_4)橄榄油的方法建立肝纤维化模型。确定造模成功后,将造模组剩余大鼠分为模型组(7只)、芪甲柔肝全方组(6只)、拆方益气养血药组(6只)、拆方化瘀通络药组(6只),芪甲柔肝全方组、拆方益气养血药组、拆方化瘀通络药组分别采用中药灌胃治疗,空白组与模型组给予生理盐水灌胃。灌胃28 d后处死大鼠,检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)判断肝功能,肝组织苏木素-伊红(HE)染色、马松(Masson)染色观察肝脏炎症及纤维化,检测肝组织α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(collagenⅠ)判断肝星状细胞激活,检测肝组织血管内皮生长因子A(VEGFA)、血管内皮生长因子受体2(VEGFR-2)、SRF、c-FOS、分化抗原簇31(CD31)、血管性血友病因子(vWF)的表达水平,扫描电镜观察肝窦内皮细胞窗孔形态以揭示各组药物抗肝纤维化的机制。结果显示,与模型组相比,芪甲柔肝方及其拆方益气养血药、化瘀通络药均能减轻肝组织病理损伤,降低肝纤维化大鼠ALT、AST和肝指数(P<0.01);各治疗药物均能降低肝组织α-SMA和collagenⅠ表达(P<0.01),降低肝纤维化大鼠肝组织CD31和vWF的表达(P<0.05),维持肝窦内皮细胞窗孔形态;各治疗药物均能降低VEGFA、SRF、c-FOS的表达(P<0.05),恢复VEGFR-2的表达(P<0.05)。其中,芪甲柔肝全方在改善大鼠肝损伤与血清肝功能,抑制肝窦毛细血管化,以及调控VEGF/SRF/c-FOS信号通路方面优于拆方的益气养血药及化瘀通络药(P<0.05)。综上,芪甲柔肝方及其拆方均能够改善肝纤维化大鼠肝功能,减轻病理损伤,抑制肝星状细胞激活,减轻肝纤维化及肝窦毛细血管化,This study aims to reveal the mechanism of Qijia Rougan Decoction(QJRG)and its disassembled formulas in mitigating hepatic fibrosis via the vascular endothelial growth factor(VEGF)/serum response factor(SRF)/c-FOS pathway and hepatic sinusoidal capillarization.Male Sprague-Dawley(SD)rats were randomized into a control group(n=6)and a modeling group(n=28).Hepatic fibrosis was induced by subcutaneous injection of 40%carbon tetrachloride(CCl4)in olive oil.The successfully modeled rats were grouped as follows:model(n=7),QJRGF(n=6),disassembled formula for benefiting Qi and nourishing blood(YQYX,n=6),and disassembled formula for resolving stasis and dredging collaterals(HYTL,n=6).The rats in the YQYX,HYTL,and QJRG groups were administrated with corresponding medicines by gavage,while those in the control group and the model group were treated by gavage of normal saline.After 28 days,the rats were sacrificed,and serum and liver samples were collected.Hematoxylin-eosin staining and Masson staining were employed to observe inflammation and fibrosis,respectively,in the liver tissue.The serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured to assess the liver function.The expression levels ofα-smooth muscle actin(α-SMA)and collagenⅠin the liver tissue were measured to evaluate the activation of hepatic stellate cells(HSCs).The expression levels of vascular endothelial growth factor A(VEGFA),vascular endothelial growth factor receptor 2(VEGFR-2),SRF,c-FOS,cluster of differentiation 31(CD31),and von willebrand factor(vWF)in the liver tissue were measured.Scanning electron microscopy(SEM)was employed to observe the morphology of the fenestrae in liver sinusoidal endothelial cells(LSECs)to elucidate the mechanisms underlying the antifibrotic effects of QJRG and its disassembled formulas.Compared with the model group,QJRG,YQYX,and HYTL mitigated the pathological changes in the liver tissue,reduced the ALT and AST levels and liver index(P<0.01),and lowered theα-SMA and collagenⅠleve

关 键 词：肝纤维化 肝窦毛细血管化 芪甲柔肝方 VEGF/SRF/c-FOS信号通路 益气养血 化瘀通络 

分 类 号：R285.5[医药卫生—中药学]