淫羊藿苷对心力衰竭血清中DCs与CD4+T细胞共培养的影响
Effect of Icariin on DCs and CD4+T Cell Differentiation in Co-Culture with Heart Failure Serum
赵玲婕 1李倩 1陈波洋 1周艳 2高洁 1柴艺汇 1陈云志1
作者信息
- 1. 贵州中医药大学,贵阳 550025
- 2. 德江县民族中医院,贵州德江 565200
- 折叠
摘要
目的 探讨淫羊藿苷(icariin,ICA)对心力衰竭血清中Dahl盐敏感大鼠骨髓源性树突状细胞(dendritic cells,DCs)与CD4+T细胞共培养中CD4+T细胞分化为辅助性T细胞(T helper cell,Th)17、调节性T细胞(T regulatory cells,Treg)的影响.方法 将50 只SPF级5周龄雄性Dahl盐敏感大鼠(模型组)及同龄10只雄性Dahl盐抵抗大鼠(正常组)喂养至18周龄后麻醉制备心力衰竭血清与正常血清;收集分离2 种大鼠的DCs与CD4+T细胞,经流式细胞仪鉴定CD4+T细胞纯度后将DCs和CD4+T细胞按照 1∶10 的比例共培养,共培养的细胞分为对照组(正常血清),模型组(心力衰竭血清),模型组+维生素D组,模型组+ICA高、中、低剂量组.药物干预 24h后收集细胞标本.流式细胞仪检测DCs和CD4+T细胞共培养中Th17、Treg的阳性表达比例;ELISA检测DCs与CD4+T细胞共培养上清液白细胞介素(interleukin,IL)-17、转化生长因子(transforming growth factor,TGF)-β1、IL-6、IL-12、IL-10 的水平.结果 ICA与维生素D的治疗结果趋势一致,均可显著降低Th17 细胞阳性表达率及Th17/Treg比值(P<0.01),升高Treg阳性表达率(P<0.01);显著升高IL-10水平(P<0.01),降低IL-17、TGF-β1、IL-6、IL-12 水平(P<0.05).结论 ICA与维生素D能通过调节机体内Th17/Treg的平衡,降低促炎细胞因子IL-17、TGF-β1、IL-6、IL-12 与升高抑炎细胞因子IL-10的表达来治疗心力衰竭.
Abstract
Objective To investigate the effect of icariin(ICA)on the differentiation of CD4+T cells into helper cell(Th)17 and regulatory cells(Treg)in co-cultures of Dahl salt-sensitive rat bone marrow-derived dendritic cells(DCs)and CD4+T cells in heart failure sera.Methods Fifty SPF-rated 5-week-old male Dahl salt-sensitive rats(model group)and 10 male Dahl salt-resistant rats of the same age(normal group)were fed to 18 weeks of age and then anesthetized to prepare heart failure serum and normal serum.DCs and CD4+T cells were collected from both rats,and DCs and CD4+T cells were co-cultured at a ratio of 1:10 after CD4+T cell purity was determined by flow cytometry.The co-cultured cells were divided into control group(normal serum),model group(heart failure serum),model group +vitamin D group(1×10-8 mol/L),and model group+icariin high、medium and low dose group.Cell specimens were collected 24 h after drug intervention.The proportion of positive expression of Th17 and Treg in DCs and CD4+T cells co-culture was detected by flow cytometry.ELISA was performed to detect interleukin(IL)-17,transforming growth factor(TGF)-β1,IL-6,IL-12 and IL-10 levels in the supernatant of DCs and CD4+T cells co-culture.Results The treatment results of ICA and vitamin D showed the same trend.Both significantly decreased Th17 cell positive expression rate and Th17/Treg ratio(P<0.01)and increased Treg positive expression rate(P<0.01),significantly increased IL-10 levels(P<0.01)and decreased IL-17,TGF-β1,IL-6 and IL-12 levels(P<0.05).Conclusion ICA and vitamin D can treat heart failure by regulating the balance of Th17/Treg in the body,decreasing the expression of pro-inflammatory cytokines IL-17,TGF-β1,IL-6,IL-12 and increasing the expression of anti-inflammatory cytokine IL-10.
关键词
树突状细胞/CD4+T细胞/心力衰竭/淫羊藿苷/维生素DKey words
Dendritic cells/CD4+T cells/Heart failure/Icariin/Vitamin D引用本文复制引用
基金项目
国家自然科学基金(81874376)
贵州省教育厅滚动支持省属高校科研平台团队项目(黔教技[2022]023)
出版年
2024