黄连解毒汤通过调节性T细胞产生抗动脉粥样硬化作用

Anti-atherosclerotic effect of a Chinese medicine,Huanglian Jiedu Decoction,mediated by regulatory T cells

目的观察黄连解毒汤(HLJDD)对ApoE^(-/-)小鼠斑块、炎症因子及调节性T细胞(Tregs)的影响。方法高脂饲料喂养雄性ApoE^(-/-)小鼠建立动脉粥样硬化(atherosclerosis,AS)模型,40只雄性ApoE^(-/-)小鼠随机分为:模型组,辛伐他汀组,HLJDD高、中、低剂量组(n=8)。HLJDD灌胃剂量依次为:每日3.5、7.0、14.0 g/(kg·bw),辛伐他汀灌胃剂量为每日5 g/(kg·bw),另设8只同龄雄性C57BL/6J小鼠为对照组,喂以普通饲料,连续给药16周后处死小鼠。检测血脂水平,观察主动脉斑块形成情况;ELISA法检测血清炎症因子水平;流式细胞术检测Tregs数量;RT-PCR法检测Foxp3 mRNA水平。结果与对照组相比,模型组中主动脉斑块明显,TC、TG与LDLC浓度显著升高,促炎因子超敏C反应蛋白(hs-CRP)、白介素-6(IL-6)与肿瘤坏死因子-α(TNF-α)水平显著升高,而HDL-C、白介素-10(IL-10)、生长转化因子-β(TGF-β)、Foxp3 mRNA水平显著降低,Tregs数量减少,二者差异有显著性(P<0.01);HLJDD能显著缩小主动脉斑块面积,降低血清TC、TG与LDL-C,并降低IL-6、hs-CRP与TNF-α水平,增加血清HDL-C、IL-10、TGF-β及Foxp3 mRNA水平,并增加Tregs数量,与模型组相比差异有显著性(P<0.01)。结论 HLJDD可显著减轻ApoE^(-/-)小鼠主动脉斑块损伤,可能与增加Tregs数量,从而降低血清炎症因子水平有关。

Objective To study the effect of a Chinese medicine,Huanglian Jiedu Decoction（ HLJDD）,on atherosclerotic plaque,inflammatory factors and regulatory T cells in ApoE^-/-mice. Methods High fat diet was given to ApoE^-/-mice to establish an atherosclerosis model. 40 male ApoE^-/-mice were randomly divided into five groups： model group,simvastatin group,low,moderate and high dose HLJDD groups（ n = 8）. HLJDD was intragastrically administered in a dose of 3. 5,7. 0,or 14. 0 g /（ kg·bw） once dailg for 16 weeks. The dose of simvastatin was 5. 0 g /（ kg·bw）. Another 8 male C57 BL /6J mice were taken as control group. At the end of the 29-week experiment,all of the mice were sacrificed. The aortic plaques,level of blood lipids,inflammatory factors,Tregs number and the level of Foxp3 mRNA were detected and analyzed by ELISA,flow cytometry and RT-PCR,respectively. Results Compared with the control group,the aortic plaques were much larger in the model group,and the levels of TC,TG,LDL-C,IL-6,hs-CRP,and TNF-α were significantly higher than those in the control group（ P〈0. 01 for all）. Meanwhile,the levels of HDL-C,IL-10,TGF-βand Foxp3 mRNA were much lower than those in the control group（ P〈0. 01 for all）,and the Tregs numbers were less than that in the control group（ P〈0. 01）. HLJDD regulated the blood lipids in ApoE^-/-mice and decreased the levels of IL-6,hs-CRP,andT NF-α,however increased the levels of IL-10,TGF-β and Foxp3 mRNA. At the same time,it increased Tregs number in the ApoE^-/-mice. Compared with the model group,the difference was statistically significancet（ P〈0. 01）. Conclusions HLJDD can significantly alleviate the aortic plaque damages in ApoE^-/-mice. It may be related to the up-regulation of Tregs,which can lead to decrease the expression of serum pro-inflammatory factors such like IL-10,hs-CRP and TNF-α.