基于胆固醇逆转运探讨泽泻白术配伍改善ApoE^(-/-)小鼠动脉粥样硬化的作用

To explore effect of Alisma combined with Atractylodes macrocephala on atherosclerosis in ApoE-/-mice with reverse cholesterol transport

目的探讨泽泻白术配伍改善ApoE-/-小鼠动脉粥样硬化(As)及其与胆固醇逆转运的相关性。方法高脂饮食喂养ApoE-/-小鼠构建As动物模型,随后以泽泻白术配伍干预。超声测量颈总动脉管壁厚度和管腔直径,HE染色检测动脉形态学改变,油红O染色观察肝脏内脂质沉积情况,ELISA检测血清肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)含量,全自动生化分析仪检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)水平,Western blot检测肝脏内的胆固醇逆转运蛋白如沉默信息调节因子1(SIRT1)、肝X受体α(LXRα)、ATP结合盒转运体A1(ABCA1)和B族Ⅰ型清道夫受体(SR-BⅠ)的表达。结果泽泻白术配伍显著改善模型鼠的颈总主动脉管壁厚度和管腔直径,抑制模型鼠血清炎症因子TNF-α、IL-1β、MMP-2和MMP-9的表达,降低TC、TG和LDL水平,增加HDL表达,促进肝脏内的胆固醇逆转运相关蛋白SIRT1、LXRα、ABCA1和SR-BⅠ表达,减轻脂质在肝脏的沉积。结论泽泻白术配伍可能通过SIRT1-LXRα-ABCA1/SR-BⅠ通路促进胆固醇逆转运,减轻肝脏内脂质沉积,改善炎症反应,抑制血管壁增厚和血管狭窄,从而发挥抗As作用。

Aim To explore the effect of the compatibility of Alisma and Atractylodes on relieving atherosclerosis(As)in ApoE-/-mice and the correlation with reverse cholesterol transport.Methods ApoE-/-mice were fed with a high-fat diet to construct an animal model of As and treated with Alisma and Atractylodes.The wall thickness,lumen diameter of common carotid artery were measured by ultrasound.HE staining was used to observe the morphology of the artery,and lipid deposition in the liver was observed by oil red O staining.Inflammatory cytokines such as tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),matrix metalloproteinase-2(MMP-2)were test by ELISA,and matrix metalloproteinase-9(MMP-9)and blood lipid components serum total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL)and high density lipoprotein(HDL)were analyzed by automatic biochemical analyser.Western blot was used to quantify the expression of silent information regulator protein 1(SIRT1),liver X receptorα(LXRα),ATP-binding cassette transporter A1(ABCA1)and scavenger receptor class B typeⅠ(SR-BⅠ)in the ApoE-/-mice liver.Results The compatibility of Alisma and Atractylodes not only significantly repressed the wall thickness,lumen diameter of the common carotid in ApoE-/-mice,but also reduced the deposition of lipids in the ApoE-/-mice liver.Treatment with Alisma decoction caused a drop in serum MMP-2,MMP-9,TNF-α,IL-1β,TC,TG,LDL and increase in serum HDL.The expression of SIRT1,LXRα,ABCA1 and SR-BⅠwere markedly up-regulated in the ApoE-/-mice liver treated with Alisma and Atractylodes.Conclusion It was suggested that the compatibility of Alisma and Atractylodes promoted reverse cholesterol transport to reduce liver lipid deposition,suppressed inflammation and improved the morphology of artery via SIRT1-LXRα-ABCA1/SR-BⅠpathway.It exerted the protective effect in the occurrence and development of As.