黄芪总皂苷、当归挥发油对动脉粥样硬化ApoE^(-/-)小鼠脂质代谢的影响

Effect of Astragalus Saponins and Angelica Volatile Oil on Lipid Metabolism in Atherosclerotic ApoE^(-/-) Mice

目的研究黄芪总皂苷、当归挥发油对动脉粥样硬化(Atherosclerosis,AS)ApoE^(-/-)小鼠脂质代谢的影响。方法将30只ApoE^(-/-)小鼠随机分为模型组、黄芪总皂苷组、当归挥发油组,每组10只,同时选用10只C57BL/6小鼠作为正常对照组。高脂饮食喂养建立动脉粥样硬化小鼠模型;采用试剂盒测定小鼠血清、肝脏总胆固醇(TC)、甘油三脂(TG)、低密度脂蛋白(LDL-c)、高密度脂蛋白(HDL-c)、内皮素(ET-1)、一氧化氮(NO)、游离脂肪酸(FFA)、游离胆固醇(FC)含量;油红O染色检测肝脏、主动脉、主动脉窦脂质沉积;HE染色观察肝脏、主动脉病理改变;以ImageJ软件统计主动脉窦脂质沉积、主动脉厚度、肝脏脂滴空洞的面积及其占总面积的百分比;Western Blot法检测脂质代谢相关蛋白的表达变化。结果高脂喂养12周后,与正常对照组比较,模型组血清TC、TG、LDL-c、HDL-c含量升高(P<0.01);与模型组比较,黄芪总皂苷、当归挥发油组血清TC、TG、LDL-c含量降低(P<0.05)。高脂喂养16周后,与正常对照组比较,模型组血清、肝脏TC、TG、LDL-c、HDL-c、ET-1、NO、FFA、FC含量升高(P<0.01);主动脉、主动脉窦脂质大量沉积;肝脏脂肪性变严重,脂滴空泡呈弥漫性分布,肝脏油红脂滴和脂滴空洞面积增多;主动脉管壁明显增厚(P<0.01),伴有大量炎症细胞浸润及胆固醇结晶沉积;肝脏ABCA1、SR-BI、APOAI、LXR-α(P<0.05)、SREBP2(P<0.01)蛋白表达降低。与模型组比较,黄芪总皂苷组、当归挥发油组血清TC、TG、LDL-c、ET-1、NO含量降低(P<0.05,P<0.01);肝脏TC、TG、FFA、FC含量降低(P<0.01);主动脉窦脂质沉积,肝脏油红脂滴及脂滴空洞面积减少(P<0.05,P<0.01),主动脉、肝脏病变相对减轻;当归挥发油组肝脏SR-BI、LXR-α(P<0.05)、ABCA1、SREBP2、APOAI(P<0.01)蛋白表达升高;黄芪总皂苷组肝脏ABCA1、SR-BI(P<0.05)、LXR-α、SREBP2、APOAI(P<0.01)蛋白表达升高。结论黄芪总皂苷、当归挥发油对动脉粥样硬化ApoE^(-/-)小鼠的脂质代谢都有一定的干预调节作用,其具体起效机制可能是通过调控ApoE^(-/-)小鼠脂质代谢相关蛋白的表达水平来实现的。

Objective To study the effect of astragalus total saponins and angelica volatile oil on lipid metabolism in atherosclerotic ApoE-/-mice.Methods Thirty ApoE-/-mice were randomly divided into model group, astragalus saponins group,and angelica volatile oil group,10 mice in each group,and 10 C57 BL/6 mice were selected as the normal control group. Here we established atherosclerosis(AS)model on ApoE^(-/-) mice induced by high-fat diet. The contents of TC,TG,LDL-c,HDL-c,ET-1,NO,FFA,FC in serum and liver were detected by kit. Oil red O staining was used to detect lipid deposits in liver, aorta, and aortic sinus. HE Staining was used to observe the pathological changes of the liver and aorta. Image J software was used to count the aortic sinus lipid deposition,aortic thickness,the area of lipid droplets in the liver and its percentage of the total area. WB was applied to detect the expression of lipid metabolism-related proteins in liver.Results After 12 weeks of high-fat feeding,the serum TC, TG, LDL-c, HDL-c levels in the model group were higher than those in the normal group(P<0.01).Compared with the model group,serum contents of TC,TG,LDL-c(P<0.05)in the astragalus total saponins and angelica volatile oil group were reduced. After 16 weeks of high-fat feeding,TC,TG,LDL-c,HDL-c,ET-1,NO,FFA,and FC levels of the serum and liver in the model group were higher than those in the normal group(P<0.01);A largea mount of lipid deposition in aortic plaque,aortic sinus was visible. Fatty liver became severe and lipid droplet vacuoles were diffusely distributed,the area of oil red lipid droplets and lipid droplets cavities in liver were increased, the wall of aorta was thickened(P<0.01), a large number of inflammatory cell infiltration and cholesterol crystal deposition appeared;Expressions of ABCA1,SR-BI,LXR-α,APOA1(P<0.05),SREBP2(P<0.01)protein in liver were reduced. Compared with the model group,the serum TC,TG,LDL-c(P<0.05),ET-1,NO(P<0.01)in the astragalus saponins and angelica volatile oil group were reduced;The contents of TC,TG,FFA,FC in liver were decreased(P<0.01);Aortic sinus lipid deposition,liver oil red lipid droplets,lipid droplet cavity area were decreased(P<0.05,P<0.01). Pathological changes of the aorta and liver were relatively reduced. Protein expressions of SR-BI、LXR-α(P<0.05)、ABCA1、SREBP2、APOAI(P<0.01)in the volatile oil group were increased. Protein expressions of ABCA1、SR-BI(P<0.05)、LXR-α、SREBP2、APOAI(P<0.01)in the angelica astragalus total saponins group were also increased.Conclusion Astragalus saponins and angelica volatile oil have a certain regulation effect on the lipid metabolism of atherosclerotic ApoE^(-/-) mice and the mechanism may be achieved by regulating the expression level of ApoE^(-/-) mouse lipid metabolism protein.