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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

周李煜, 马元婧, 孙玉婷, 陈韦凯, 邱润泽, 袁冬平, 龙军

周李煜, 马元婧, 孙玉婷, 陈韦凯, 邱润泽, 袁冬平, 龙军. 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[J]. 中国药科大学学报, 2020, 51(1): 60-67. DOI: 10.11665/j.issn.1000-5048.20200110
引用本文: 周李煜, 马元婧, 孙玉婷, 陈韦凯, 邱润泽, 袁冬平, 龙军. 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[J]. 中国药科大学学报, 2020, 51(1): 60-67. DOI: 10.11665/j.issn.1000-5048.20200110
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ZHOU Liyu, MA Yuanjing, SUN Yuting, CHEN Weikai, QIU Runze, YUAN Dongping, LONG Jun. Effects of Yangyin Huoxue Prescription on the imbalance of Treg/Th17 and atherosclerosis in ApoE-/- mice induced by high-fat diet[J]. Journal of China Pharmaceutical University, 2020, 51(1): 60-67. DOI: 10.11665/j.issn.1000-5048.20200110
Citation: ZHOU Liyu, MA Yuanjing, SUN Yuting, CHEN Weikai, QIU Runze, YUAN Dongping, LONG Jun. Effects of Yangyin Huoxue Prescription on the imbalance of Treg/Th17 and atherosclerosis in ApoE-/- mice induced by high-fat diet[J]. Journal of China Pharmaceutical University, 2020, 51(1): 60-67. DOI: 10.11665/j.issn.1000-5048.20200110
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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

  • 南京中医药大学药学院

基金项目: 国家自然科学基金资助项目(No.81973823,No.81573929)

Effects of Yangyin Huoxue Prescription on the imbalance of Treg/Th17 and atherosclerosis in ApoE-/- mice induced by high-fat diet

摘要

    摘要
  • 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/kg，以生药计)、YHF高剂量(36 g/kg，以生药计)灌胃给药20周进行干预。油红O脂肪染色法观测主动脉斑块面积变化；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。
    Abstract: Atherosclerosis(AS), characterized with the accumulation of lipids on the vessel wall, is an immune-related inflammatory disease which promotes the progression of cardiovascular diseases(CVD). The imbalance of Treg/Th17 accelerates the progression of AS. Yangyin Huoxue Prescription(YHF)is an efficient traditional Chinese medicine used in the treatment of AS, but the effects of YHF on the balance of immunity have still not been clarified. 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 given HFD to induce AS and administered low-dose YHF(18 g/kg)or high-dose 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, and then treated with lipopolysaccharide(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 lipid 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 decreasing 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.
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  • 刊出日期:  2020-02-24

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