DAPT调节Treg/Th17细胞免疫平衡抑制动脉粥样硬化

DAPT regulates Treg/Th17 cells immune balance to inhibit atherosclerosis

目的:观察Notch信号抑制剂分泌酶抑制剂(γ-DAPT)对动脉粥样硬化小鼠病理改变及Treg/Th17细胞免疫平衡的影响。方法:将24只Apo E基因敲除C57BL小鼠随机分为空白组、模型组和DAPT组。空白组用普通饲料饲养,模型组和DAPT组用高脂饲料饲养。饲养5周后,DAPT组小鼠以100 mg/(kg·d)皮下注射DAPT(溶于DMSO中),其余两组皮下注射等量DMSO。5周后,采用病理染色分析各组小鼠动脉病理改变,ELISA检测血浆中IL-17水平,采用流式细胞术检测各组小鼠脾脏Treg/Th17细胞比例。结果:HE染色结果显示,模型组有明显粥样斑块形成和泡沫细胞形成,DAPT组动脉病变程度比粥样动脉硬化模型组明显减轻。正常组、模型组和DAPT组各组血浆中IL-17水平分别为(293.94±28.59)、(454.05±172.68)和(335.40±89.57)pg/ml;DAPT降低AS小鼠血浆IL-17水平(P<0.05)。空白组、模型组和DAPT组各组小鼠Treg细胞百分比分别为(3.80±0.56)%、(2.54±0.38)%和(4.73±0.64)%;DAPT降低AS小鼠血浆IL-17水平(P<0.05)。空白组、模型组和DAPT组各组小鼠Th17细胞亚群分别为(3.46±0.23)%、(4.52±0.85)%和(1.38±0.37)%。结论:DAPT降低AS小鼠血浆IL-17的水平,抑制Th17细胞亚群分化,而促进Treg细胞分化,通过改变Treg/Th17细胞免疫平衡从而减轻动脉粥样硬化。

Objective： To observe the effect of Notch signal inhibitor DAPT（ γ-secretase inhibitor） on the pathological changes of atherosclerosis mice and the immune balance of Treg / Th17. Methods： 24 Apo E knockout C57 BL mice were randomly divided into blank group,model group and DAPT group. The blank group were fed with normal diet,the model group and the experimental group were fed with high fat diet. After 5 weeks of feeding,the mice in the experimental group were injected with DAPT[100 mg /（ kg·d）,resuspended in DMSO],and the other two groups were injected with the equivalent amount of DMSO. After another 5 weeks,pathological changes of the mice in each group were analyzed by HE staining. ELISA was used to detect the level of IL-17 in plasma,and the proportion of splenic Treg / Th17 cells in each group was detected by flow cytometry. Results： HE staining results showed that the model group had obvious plaque formation and foam cell formation,which showed that the AS model was successfully prepared. The degree of arterial disease in the DAPT group was significantly less than that in the model group. The plasma levels of IL-17 in the blank group,model group and DAPT group were（ 293. 94 ± 28. 59）,（ 454. 05 ± 172. 68） and（ 335. 40 ± 89. 57） pg / ml,respectively. The percentages of Treg cells in the blank group,model group and DAPT group were（ 3. 80± 0. 56） %,（ 2. 54± 0. 38） % and（ 4. 73± 0. 64） %,respectively. The Th17 cell subsets of mice in the blank group,model group and DAPT group were（ 3. 46 ± 0. 23） %,（ 4. 52 ± 0. 85） % and（ 1. 38±0. 37） %,respectively. Conclusion： DAPT decreased the plasma level of IL-17 in AS mice,inhibited the differentiation of Th17 cell subsets,and promoted the differentiation of Treg,and reduced the atherosclerosis by changing the Treg / Th17 cells immune balance.