基于Th17/Treg轴探讨健脾养肝汤治疗自身免疫性肝炎的分子机制  

Exploration of molecular mechanism of Jianpi Yanggan Decoction in treating autoimmune hepatitis based on Th17/Treg axis

作　　者：于盈盈 孙婷婷[1] 杨润萌 张亚 包一珺 朱丹[1] YU Yingying;SUN Tingting;YANG Runmeng;ZHANG Ya;BAO Yijun;ZHU Dan(Department of Infectious Diseases,Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China)

机构地区：[1]中国中医科学院广安门医院感染疾病科,北京100053

出　　处：《长春中医药大学学报》2024年第12期1330-1334,共5页Journal of Changchun University of Chinese Medicine

基　　金：国家重点研发计划项目(2018YFC1705700);中国中医科学院科技创新工程重大攻关项目(CI2021A00809)。

摘　　要：目的基于Th17/Treg轴探讨健脾养肝汤治疗自身免疫性肝炎分子机制。方法将Balb/c小鼠随机分为对照组、模型组、泼尼松龙组、健脾养肝汤组。造模方法采用尾静脉注射刀豆蛋白A(ConA),每周1次,连续6周。同时分别给予相应药物灌胃治疗,每日1次,连续6周。电镜扫描下观察各组肝脏组织超微结构差异。流式细胞术检测脾脏和外周血Th17、Treg细胞亚群比例,流式荧光法检测血浆内炎性细胞因子(IL-2、IL-6、IL-17A、TNF-α、IL-4、IL-10)水平,免疫组化法检测肝脏组织转录因子RORγt、Foxp3表达。结果电镜观察,对照组肝细胞索和肝窦结构清晰,肝窦内皮细胞表面光滑,其上窗孔网均匀分布。模型组小鼠肝细胞索和肝窦结构及窗孔网模糊紊乱,泼尼松龙组与健脾养肝汤组肝窦和窗孔网结构相对清晰。脾脏和外周血Th17、Treg细胞,与对照组比较,模型组脾脏、外周血Th17细胞均明显升高,外周血Treg细胞明显降低(P<0.05),脾脏Th17/Treg细胞比值明显上升(P<0.01);与模型组比较,泼尼松龙组与健脾养肝汤组脾脏Th17细胞、Th17/Treg细胞比值均明显降低(P<0.01)。血浆炎性细胞因子比较,与对照组比较,模型组IL-17水平明显升高(P<0.05),IL-10明显降低(P<0.05);与模型组比较,健脾养肝汤组IL-17明显降低(P<0.05)。免疫组化结果:与对照组相比,模型组RORγt表达增加;与模型组比较,泼尼松龙组与健脾养肝汤药物组的RORγt表达减少。结论健脾养肝汤可能通过调控转录因子RORγt、Foxp3表达,调节Th17/Treg轴平衡,降低炎症因子表达,进而改善肝窦结构和肝窦内皮细胞免疫微环境,减轻肝脏炎症损伤。Objective To explore the mechanism of action of Jianpi Yanggan Decoction in treating autoimmune hepatitis based on helper T cell 17(Th17)/regulatory T cells(Treg)axis.Methods Balb/c mice were randomly divided into 4 groups,namely the control group,the model group,Prednisolone group,and Jianpi Yanggan Decoction group.The modeling method was performed by injecting Concanavalin A(ConA)into the tail vein once a week for 6 consecutive weeks.At the same time,corresponding drugs were given by gavage once a day for 6 consecutive weeks.The differences in the ultrastructure of liver tissues among different groups under electron microscopy scanning were observed.Flow cytometry was used to detect the ratios of Th17 and Treg cell subsets in the spleen and peripheral blood.Flow fluorescence assay was used to detect the levels of inflammatory cytokines[interleukin(IL)-2,IL-6,IL-17A,tumor necrosis factor-α(TNF-α),IL-4,IL-10]in plasma.And immunohistochemistry was used to detect the expression of transcription factors RORγt and Foxp3 in liver tissue.Results Electron microscopy observation:In the control group,hepatocyte cords and the structue of hepatic sinusoids were clear,and surface of liver sinuysoidal endothelial cells was smooth,with a uniform distribution of suprafenestral networks.In the model group,hepatocyte cords,the structure of hepatic sinusoids,and fenestral network were obviously blurred and disordered,while in Prednisolone group and Jianpi Yanggan Decoction group,the structure of hepatic sinusoids,and fenestral network were relatively clear and distinct.Spleen and peripheral blood Th17 and Treg cells:Compared with the control group,the model group showed a significant increase in Th17 cells in the spleen and peripheral blood,a significant decrease in Treg cells in peripheral blood(P<0.05),and a significant increase in the ratio of Th17/Treg cells in the spleen(P<0.01).Compared with model group,the spleen Th17 cells and Th17/Treg cell ratio were significantly decreased in Prednisolone group and Jianpi Yanggan Dec

关 键 词：健脾养肝汤 自身免疫性肝炎 Th17/Treg轴 肝脏免疫微环境 

分 类 号：R256.4[医药卫生—中医内科学]