温阳化浊通络方对硬皮病模型小鼠皮肤Th17细胞及相关细胞因子的影响  

Effects of Wenyang Huazhuo Tongluo Formula on Th17 cells and related cytokines in skin of mice with scleroderma

作　　者：郭嘉 韩立 单雨 陈爽[1] 刘畅[1] 卞博 卞华 GUO Jia;HAN Li;SHAN Yu;CHEN Shuang;LIU Chang;BIAN Bo;BIAN Hua(Henan University of Chinese Medicine,Zhengzhou 450000,China;ZHANG Zhong-jing School of Chinese Medicine,Nanyang Institute of Technology,Henan Key Laboratory of ZHANG Zhong-jing Formulae and Herbs for Immunoregulation,Nanyang 473004,China)

机构地区：[1]河南中医药大学,郑州450000 [2]南阳理工学院张仲景国医国药学院,河南省张仲景方药与免疫调节重点实验室,南阳473004

出　　处：《中华中医药杂志》2023年第3期1259-1264,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81774300,No.82074415)。

摘　　要：目的:探讨温阳化浊通络方对硬皮病模型小鼠皮肤T细胞辅助细胞17(Th17)及相关细胞因子的影响。方法:将60只小鼠随机分为空白组、模型组、miR-155antagomir抑制剂组(250 nmol/3 d)和温阳化浊通络方组(低、中、高剂量分别为21、42、84 g·kg^(-1)·d^(-1)),每组10只。使用盐酸博来霉素建立硬皮病模型。连续给药3周后,取小鼠皮肤组织进行HE染色、应用免疫组化检测白细胞介素(IL)-6和IL-17含量、应用流式细胞术检测Th17细胞比例、应用实时荧光定量PCR(qPCR)检测皮肤miR-155及IL-6、IL-17 mRNA的相对表达量。结果:S S C模型小鼠皮肤炎症水平远高于正常小鼠。免疫组化显示,硬皮病模型组I L-6、I L-17含量高于正常小鼠(P<0.01);温阳化浊通络方可有效降低IL-6、IL-17含量(P<0.01)。模型组皮肤CD4+IL17+Th17细胞比例,miR-155、IL-6及IL-17 mRNA相对表达量显著高于空白组(P<0.01,P<0.05),温阳化浊通络方中、高剂量组可有效降低上述指标(P<0.01)。结论:温阳化浊通络方可能通过抑制硬皮病模型小鼠皮肤miR-155、IL-6、IL-17基因表达,进而调控Th17细胞,影响皮肤成纤维细胞增殖,减缓硬皮病模型小鼠皮肤纤维化进程。Objective:To explore the effects of Wenyang Huazhuo Tongluo Formula(WYTL)on T cell helper cell 17(Th17)and related cytokines in the skin of mice with scleroderma.Methods:Sixty mice were randomly divided into blank group,model group,miR-155 antagomir inhibitor group(250 nmol/3 d)and WYTL group(low-,medium-and high-dose were 21,42,84 g·kg^(-1)·d^(-1)),with 10 mice in each group.Bleomycin hydrochloride was used to establish scleroderma model.After 3 weeks of continuous administration,the skin tissues of mice were stained with HE,interleukin-6(IL-6)and interleukin-17(IL-17)were detected by immunohistochemistry,the proportion of Th17 cells was detected by flow cytometry,and the relative expressions of miR-155,IL-6 and IL-17 mRNA in skin were detected by real-time fluorescence quantitative PCR(qPCR).Results:The level of skin inflammation in scleroderma model mice was much higher than that in normal mice.Immunohistochemistry showed that the contents of IL-6 and IL-17 in scleroderma model group were higher than those in normal mice(P<0.01).WYTL could effectively reduce the content of IL-6 and IL-17(P<0.01).The proportion of skin CD4+IL17+Th17 cells and the relative expression levels of miR-155,IL-6 and IL-17 mRNA in model group were significantly higher than those in model group(P<0.01,P<0.05).The medium-and high-dose groups of WYTL could effectively reduce the above indexes(P<0.01).Conclusion:WYTL may regulate Th17 cells,affect the proliferation of skin fibroblasts and slow down the process of skin fibrosis in scleroderma model mice by inhibiting the expression of miR-155,IL-6 and IL-17 genes in skin.

关 键 词：温阳化浊通络方 硬皮病 动物模型 TH17细胞 免疫组化 

分 类 号：R285.5[医药卫生—中药学]