摘要
目的观察滋肾通络方(ZTF)对小鼠Th17细胞分化和活性的干预作用,探讨其抗类风湿关节炎的可能机制。方法制备ZTF冻干粉及含药血清,磁珠分选法提取C57BL/6小鼠脾细胞中的CD4^+T细胞进行体外培养,采用CCK-8法筛选ZTF冻干粉及含药血清对CD4^+T细胞活力影响的最佳浓度;另取体外培养的CD4^+T细胞,随机分为空白对照组、ZTF冻干粉0.5、1、2 mg/mL组、普通大鼠血清对照组、ZTF含药血清5%、10%、15%浓度组。ELISA法检测细胞上清IL-17含量;q-PCR检测IL-17、维甲酸相关核孤儿受体γt(ROR-γt)mRNA表达水平。结果与对照组比较,ZTF冻干粉各剂量组和含药血清10%、15%浓度组细胞上清IL-17水平降低(P<0.05),ZTF冻干粉2 mg/mL组及含药血清各浓度组IL-17 mRNA水平降低(P<0.05),同时ZTF冻干粉1、2 mg/mL组及含药血清不同浓度组细胞ROR-γt mRNA水平降低(P<0.05)。结论ZTF冻干粉及其含药血清均可抑制Th17细胞的分化和活性,提示其活性成分具有对Th17细胞的直接干预作用,这可能是其防治类风湿关节炎的机制之一。
Objective To observe the effects of Zishen Tongluo Formula(ZTF)on Th17 cells and further elucidate the potential mechanism of ZTF on rheumatoid arthritis(RA).Methods After preparation of ZTF freeze-dried powder and drug-containing serum,CD4^+T cells were isolated from splenocytes of C57 BL/6 mice by magnetic cell sorting with mouse anti-CD4 microbeads.The effects of ZTF freeze-dried powder and drug-containing serum on CD4^+T cell viability were evaluated by the CCK8 method to determine the optimal concentrations.Besides,cultured CD4^+T cells in vitro were randomly divided into the control group,the 0.5,1 and 2 mg/mL ZTF freeze-dried powder groups,the normal rat serum group,the 5%,10%and 15%ZTF containing serum groups.The level of IL-17 was detected by ELISA.The expression of IL-17 mRNA and ROR-γt mRNA were measured by q-PCR.Results Compared with the control group,ZTF freeze-dried powder,10%and 15%ZTF containing serum markedly decreased the levels of IL-17 in the supernatants(P<0.05).The expression of IL-17 mRNA were significantly inhibited by 2 mg/mL ZTF freeze-dried powder and ZTF containing serum groups,together with the low levels of ROR-γt mRNA in 1,2 mg/mL ZTF freeze-dried powder groups and ZTF containing serum groups(P<0.05).Conclusions Both ZTF freeze-dried powder and its drug-containing serum could inhibit the differentiation and activity of Th17 cells.It suggested that the active ingredients of ZTF have direct effects on Th17 cells,which may be one of its mechanisms for RA treatment.
作者
杨培
范瑶婕
张明菲
王祥
周玲玲
YANG Pei;FAN Yao-jie;ZHANG Ming-fei;WANG Xiang;ZHOU Ling-ling(Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica,College of Pharmacy,Nanjing University of Chinese Medicine Nanjing,210023)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2020年第8期967-971,共5页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金面上项目(No.81673937)
江苏省研究生科研与实践创新计划(No.KYCX19_1326)。
关键词
滋肾通络方
类风湿关节炎
冻干粉
含药血清
TH17细胞
Zishen Tongluo Formula
rheumatoid arthritis
freeze-dried powder
drug-containing serum
Th17 cells
