芪术颗粒对肝窦内皮细胞的影响及机制研究  

Effect of Qizhu Granule on hepatic sinusoidal endothelial cells and its mechanism

作　　者：姜心航 张荣 刘绍能[1] 马继征[1] 丁佳媛 刘慧敏[1] 张玲[3] 路迎冬[3] JIANG Xinhang;ZHANG Rong;LIU Shaoneng;MA Jizheng;DING Jiayuan;LIU Huimin;ZHANG Ling;LU Yingdong(Guang'anmen Hospital,Chinese Academy of traditional Chinese Medicine,Beijing 100053)

机构地区：[1]中国中医科学院广安门医院消化内科,北京100053 [2]新疆维吾尔自治区人民医院中医科 [3]中国中医科学院广安门医院病理科

出　　处：《环球中医药》2022年第8期1343-1349,共7页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81573917,81973832)。

摘　　要：目的 探讨芪术颗粒含药血清对四氯化碳(CCl)诱导的肝纤维化模型大鼠肝窦内皮细胞(liver sinusoidal endothelial cells,LSECs)失窗孔化及整合素αVβ3(integrin αVβ3,ITGαVβ3)-黏着斑激酶(focal adhesion kinase,FAK)-大鼠肉瘤(rat sarcoma,Ras)/丝裂原活化蛋白激酶(mitogen-activated protein kinases, MAPK)信号通路的影响与机制。方法 70只雄性Wistar大鼠,先取50只大鼠分为正常对照组、模型组及芪术颗粒高、中、低剂量组,每组10只,分别给予相应处理,制备含药血清和正常对照组血清。另20只雄性Wistar大鼠用于制备肝窦内皮细胞,其中14只大鼠采用CCl腹腔注射建立肝纤维化模型4周后,采用Masson与HE染色观察肝纤维化程度。另6只大鼠作为正常对照。分离LSECs,将LSECs分为阳性对照组、模型组、芪术低剂量组、芪术中剂量组、芪术高剂量组,其中模型组加入正常大鼠血清,其余组分别加入相应含药血清,常规培养48小时,进行相关指标检测。流式细胞仪检测大鼠肝窦内皮细胞中的血小板-内皮细胞黏附分子(platelet endothelial cell adhesion molecule-1,CD31)阳性细胞比例和肝窦内皮细胞1抗体(hepatic sinusoidal endothelial cells antibody,SE-1)阳性细胞比例;扫描电镜观察肝窦内皮细胞窗口情况;Western Blot法检测LSECs中CD31、SE-1、Ras、ITGαVβ3、FAK、MAPK、磷酸化黏着斑激酶(phosphorylation focal adhesion kinase,p-FAK)及磷酸化丝裂原活化蛋白激酶(phosphorylation mitogen-activated protein kinases,p-MAPK)蛋白表达情况。结果 (1)大鼠CCl腹腔注射造模后纤维增生显著,肝纤维化LSECs组的CD31阳性细胞比例和SE-1阳性细胞比例均显著升高。(2)模型组窗孔数少而稀疏,芪术颗粒干预后,扫描电镜观察LSECs窗孔数有所恢复,尤以芪术高剂量组明显。(3)与模型组比较,芪术各剂量组的SE-1、CD31、αVβ3表达量明显降低(P<0.05),其中芪术各剂量组的SE-1、CD31、αVβ3表达量随剂量的�Objective To investigate the effects of Qizhu granule containing serum on fenestration and integrin of hepatic sinusoidal endothelial cells( LSECs) in carbon tetrachloride( CCl)-induced hepatic fibrosis model rats αVβ3-FAK Ras/MAPK signaling pathway.Methods 70 male Wistar were prepared for this experientment. 50 rats were divided into the normal control group, the model group and the Qizhu granule high, medium and low dose groups, with 10 rats in each group. They were treated respectively to prepare drug containing serum and the normal control group serum. The other 20 rats were used to prepare hepatic sinusoidal endothelial cells. Among them,14 rats were injected intraperitoneally with CClto establish hepatic fibrosis model. After 4 weeks, the degree of hepatic fibrosis was observed by the methods of Masson and HE staining. The other 6 rats were used as the normal control. Lsecs were separated and divided into the positive control group, model group, low dose group, medium dose group and high dose group. The model group was added with normal rat serum, and the other groups were added with corresponding drug containing serum. After 48 hours of routine culture, the relevant indexes were detected. The proportion of platelet endothelial cell adhesion molecule-1(CD31) positive cells andhepatic sinusoidal endothelial cells antibody(SE-1) positive cells in rat hepatic sinusoidal endothelial cells were detected by flow cytometry,the window of hepatic sinusoidal endothelial cells was observed by scanning electron microscope, and CD31, SE-1,rat sarcoma( Ras) and integrin αVβ3,( ITGαVβ3). Expression offocal adhesion kinase(FAK), mitogen-activated protein kinases(MAPK), phosphorylation focal adhesion kinase(p-FAK) andphosphorylation mitogen-activated protein kinases(p-MAPK) protein.Results (1)After intraperitoneal injection of CClin rats, fibrogenesis was improved. The proportion of CD31 positive cells and SE-1 positive cells in the hepatic fibrosis lsecs group was increased significantly. 2. The number of fenestras i

关 键 词：肝纤维化 肝窦内皮细胞 芪术颗粒 窗孔 动物实验 细胞实验 信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]