摘要
目的探讨四君子汤对高脂诱导ApoE^-/-小鼠致动脉粥样硬化(atherosclerosis,AS)主动脉线粒体能量代谢的影响。方法将24只ApoE^-/-小鼠随机分为模型组、四君子汤组和辛伐他汀组,8只C57BL/6 Cnc小鼠作为正常组。除正常组给予基础饲料外,其余各组给予高脂饲料。造模12周后,灌胃给药,正常组与模型组给予相应体积的生理盐水,四君子汤组与辛伐他汀组给予相应药物。4周后全自动生物化学分析仪检测血清甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)和低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)的含量;HE和油红O染色观察主动脉病理及斑块变化情况;采用透射电镜观察主动脉线粒体超微结构的变化;采用紫外吸收法检测线粒体呼吸链酶复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ酶活力的影响;采用Wes全自动蛋白质印迹定量分析系统检测线粒体能量代谢相关蛋白Cytc、ATP5a、Ndufs1、Cox5a的表达。结果与正常组比较,模型组小鼠血清TC、TG和LDL-C含量显著上升(P<0.01),HDL-C含量无明显变化;HE染色结果显示小鼠主动脉管腔中斑块面积范围较大、程度较重;油红O结果显示小鼠主动脉脂质沉积明显;电镜结果显示小鼠主动脉线粒体形状不规则,膜缺损,基质变淡,嵴模糊或者断裂甚至消失;小鼠主动脉线粒体呼吸链复合体酶Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ酶活力明显降低(P<0.01);模型组小鼠主动脉ATP5a、Ndufs1、Cox5a蛋白表达降低,Cytc蛋白表达升高(P<0.01)。与模型组比较,辛伐他汀组、四君子汤组TG、TC、LDL-C水平显著降低(P<0.05,P<0.01),HDL-C含量无明显变化;辛伐他汀组和四君子汤组HE染色结果显示小鼠主动脉管腔中斑块面积范围明显减小、程度明显减轻;小鼠主动脉油红O染色结果显示脂质沉积不同程度降低;电镜结果显示小鼠主动脉线粒体形状较规则
Objective To investigate the effect of Sijunzi Decoction on mitochondrial energy metabolism in aorta of atherosclerosis(AS)mice induced by ApoE^-/-.Methods Twenty-four ApoE^-/-mice were randomly divided into model group,Sijunzi Decoction group and simvastatin group,and 8 C57 BL/6 Cnc mice were selected as normal group.Except the normal group,the other groups were given high-fat feed.After 12 weeks of modeling,the drug was administered by gavage.Normal group and model group were given corresponding volume of normal saline,and Sijunzi Decoction group and simvastatin group were given corresponding drugs.Serum levels of triglyceride(TG),total cholesterol(TC),high-density lipoprotein(HDL-C)and low-density lipoprotein(LDL-C)were measured by automatic biochemical analyzer after 4 weeks.HE and oil red O staining were used to observe aortic pathology and plaque changes.The ultra-structure of aortic mitochondria was observed by transmission electron microscopy.Ultraviolet absorption method was used to detect influence on the mitochondrial respiratory chain enzyme complexesⅠ,Ⅱ,Ⅲ,ⅣandⅤ.The expressions of mitochondrial energy metabolism related proteins Cytc,ATP5 a,Ndufs1 and Cox5 a were detected by Wes automatic Western Blot quantitative analysis system.Results Compared with those of the normal group,the serum levels of TC,TG and LDL-C in the model group increased significantly(P<0.01),and HDL-C did not change significantly.HE staining results showed that the plaque area in the main vascular cavity of mice was larger and more severe.The results of oil red O showed significant lipid deposition in the aorta of mice.The results of electron microscopy showed irregular shape of aortic mitochondria,membrane defect,matrix thinning,cristae blurred or broken or even disappeared.The activities of mitochondrial respiratory chain complex enzymesⅠ,Ⅱ,Ⅲ,ⅣandⅤof the aorta decreased obviously(P<0.01).The expression levels of ATP5 a,Ndufs1 and Cox5 a in the aorta of the model group were decreased,while the expression level
作者
王莹
宋囡
冷雪
杜莹
吕美君
吴瑾
贾连群
WANG Ying;SONG Nan;LENG X ue;DU Ying;LYU Meijun;WU Jin;JIA Lianqun(Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)
出处
《中华中医药学刊》
CAS
北大核心
2020年第8期174-178,I0033,I0034,共7页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金(81803986)
辽宁省自然科学基金(20180550767,2019-ZD-0966)
沈阳市科技计划(18-013-0-73)。
关键词
四君子汤
AS
线粒体能量代谢
主动脉
Sijunzi Decoction
atherosclerosis
mitochondrial energy metabolism
aorta
