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养阴活血方对高脂饮食诱导的ApoE-/-小鼠Treg/Th17失衡及动脉粥样硬化的影响
投稿时间:2019-11-15  修订日期:2019-12-19  点此下载全文
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作者单位E-mail
周李煜 南京中医药大学 liyuzhou210@sina.com 
马元婧 南京中医药大学  
孙玉婷 南京中医药大学  
陈韦凯 南京中医药大学  
邱润泽 南京中医药大学  
袁冬平 南京中医药大学 annieyuan99@163.com 
龙军 南京中医药大学  
基金项目:国家自然科学基金项目(面上项目,重点项目,重大项目)
中文摘要:探讨养阴活血方(YHF)对高脂饮食诱导的ApoE-/-小鼠Treg/Th17失衡及动脉粥样硬化(Atherosclerosis,AS)的影响。小鼠给予高脂饮食诱导AS,YHF低剂量(18 g生药/kg)、YHF高剂量(36 g生药/kg)灌胃给药20周进行干预。油红O染色观测主动脉斑块;生化分析血脂水平;流式细胞术检测外周血Treg、Th17比例;荧光实时定量PCR(qRT-PCR)、免疫组化(IHC)及Western blot(WB)检测主动脉Foxp3、RORγt mRNA及蛋白表达。分离小鼠脾脏CD4 T细胞并用anti-CD3/CD28活化,再经LPS刺激和YHF干预后,qRT-PCR与免疫荧光(IF)检测CD4 T细胞Foxp3、RORγt表达。结果显示,YHF显著减小主动脉斑块面积,降低血脂水平,增加Treg/Th17比值;上调主动脉Foxp3表达而下调RORγt表达;且在体外上调CD4 T细胞 Foxp3表达而下调RORγt表达。本研究表明,YHF可以调节高脂饮食诱导的ApoE-/-小鼠Treg/Th17比例失衡,减弱LPS对CD4 T细胞的炎性刺激影响,从而改善AS。
中文关键词:动脉粥样硬化  ApoE-/-小鼠  养阴活血方  调节性T细胞  辅助性T细胞17
 
Effects of YHF on the Imbalance of Treg/Th17 and Atherosclerosis in ApoE-/- Mice Induced by High-fat Diet
Abstract:Atherosclerosis (AS), characterized with the accumulation of lipids in the vessel, is an immune-related inflammatory disease which promoted the progression of cardiovascular diseases (CVD). The imbalance of Treg/Th17 accelerated the progression of AS. Yangyin Huoxue Prescription (YHF) is an efficient traditional Chinese medicine used in AS treatment but the effects of YHF on the balance of immunity have not been researched. This project was designed to investigate the effects of YHF on the imbalance of Treg/Th17 and AS in ApoE-/- mice induced by high-fat diet (HFD). ApoE-/- mice were administrated HFD to induce AS and administrated low-dose of YHF (18 g/kg) or high-dose of YHF (36 g/kg) for 20 weeks. Atherosclerotic plaque area was analyzed by oil red O staining. Serum lipids were measured by biochemical kits. Treg or Th17 cells in peripheral blood were detected by flow cytometry. mRNA and protein expression of Foxp3 and RORγt of aortas were determined by qRT-PCR, western blot and immunohistochemistry. Splenic CD4 T cells of mice were isolated and activated by anti-CD3/CD28, then treated with LPS and YHF. The expression of mRNA and protein of Foxp3 and RORγt were detected by qRT-PCR and immunofluorescence. It was found that YHF reduced the plaque area, decreased lipids level and increased the ratio of Treg cells in peripheral blood. Moreover, YHF increased mRNA or protein expression of Foxp3 in aortas in vivo or CD4 T cells in vitro while decreased mRNA or protein expression of RORγt. These results suggested that YHF can regulate the imbalance of Treg/Th17 in ApoE-/- mice induced by HFD, and reduce the inflammatory stimulation of LPS on CD4 T cells, thereby improving AS.
keywords:Atherosclerosis  ApoE-/- mice  YHF  Treg  Th17
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