心脉康增强血管平滑肌细胞自噬改善ApoE^(-/-)小鼠动脉粥样硬化  

Xinmaikang Improves Atherosclerosis in ApoE^(-/-)Mice by Enhancing Autophagy of Vascular Smooth Muscle Cells

作　　者：陈鑫[1,2,3,4] 叶小汉 曹艳红 郭依宁 庄皓文 王陵军 冼绍祥[2,3,4] 王婷[1] CHEN Xin;YE Xiaohan;CAO Yanhong;GUO Yining;ZHUANG Haowen;WANG Lingjun;XIAN Shaoxiang;WANG Ting(Dongguan Hospital,Guangzhou University of Chinese Medicine,Dongguan 523127 Guangdong,China;The First Afiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The First Clinical Medical School,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学东莞医院,广东东莞523127 [2]广州中医药大学第一附属医院,广东广州510405 [3]广州中医药大学第一临床医学院,广东广州510405 [4]广州中医药大学岭南医学研究中心,广东广州510405

出　　处：《中药新药与临床药理》2023年第7期914-920,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省基础与应用基础研究基金青年基金项目(2020A1515110132);叶小汉东莞市名中医药专家传承工作室。

摘　　要：目的探讨心脉康(鳖甲、三棱、莪术等组成)调节血管平滑肌细胞(VSMC)自噬改善动脉粥样硬化(AS)的作用机制。方法72只敲除载脂蛋白E基因(ApoE^(-/-))的小鼠随机分为空白组、模型组、阳性药物(阿托伐他汀钙片)对照组(2.6 mg·kg^(-1))及心脉康低、中、高(30、60、90 g·kg^(-1))剂量组,每组12只。空白组小鼠喂养常规饮食,余下各组喂养高脂饮食12周,建立AS模型。模型复制成功后给予心脉康灌胃6周,每天1次。检测小鼠血清血脂四项:总胆固醇(Total cholesterol,TC)、甘油三酯(Triglyceride,TG)、高密度脂蛋白胆固醇(High density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(Low density lipoprotein cholesterol,LDL-C);油红O、HE和Masson染色观察小鼠主动脉根部血管组织病理变化;免疫荧光检测主动脉根部血管组织微管相关蛋白l轻链3B(LC3B)和平滑肌22 alpha蛋白(SM22α)的表达;采用Western Blot法检测主动脉血管组织中LC3B、自噬效应蛋白1(Beclin-1)和SM22α蛋白的表达水平;qRT-PCR法检测主动脉中LC3B、Beclin-1和SM22αmRNA的表达。结果与空白组比较,模型组小鼠血清血脂四项结果显示,HDL-C下降,TC、TG、LDL-C均升高(P<0.05);油红O、HE和Masson染色结果显示,小鼠主动脉根部血管组织斑块面积增多,内壁增厚并伴有大量胆固醇结晶以及炎症细胞浸润,胶原纤维含量减少;免疫荧光结果显示,LC3B和SM22α共定位面积明显减少(P<0.05);qRT-PCR和Western Blot结果显示,LC3B,Beclin-1和SM22α蛋白及m RNA表达下降(P<0.05)。与模型组比较,心脉康低、中、高剂量组及阳性药物对照组HDL-C水平明显升高,TC、TG、LDL-C水平明显降低(P<0.05);主动脉内斑块面积减少,胆固醇结晶以及炎症浸润降低,胶原纤维含量增多;LC3B和SM22α共定位面积明显增加(P<0.05);LC3B、Beclin-1、SM22α蛋白以及mRNA表达增加(P<0.05)。结论心脉康能降低血脂水平,减少斑块面积,改善动脉粥样硬化Objective To investigate the mechanism of Xinmaikang(composed of Trionycis Carapax,Sparganii Rhizoma,Curcumae Rhizoma,etc.)on autophagy regulation in vascular smooth muscle cell(VSMC)to ameliorate atherosclerosis(AS).Methods Seventy-two ApoE^(-/-)knockout mice were randomly divided into blank group,model group,positive drug group(Atorvastatin Calcium Tablets,2.6 mg·kg^(-1)),Xinmaikang low-,medium-and highdose groups(30,60,90 g·kg^(-1)),with 12 mice in each group.Mice in the blank group were fed a conventional diet,and the other groups were fed a high-fat diet for 12 weeks to establish the AS model.After the model was successfully replicated,Xinmaikang was orally administered daily for 6 weeks,total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C)and low density lipoprotein cholesterol(LDL-C)were detected.Oil red O,HE and Masson staining were used to observe the pathological changes of blood vessels at the aortic root.The expressions of microtubule-associated protein l light chain 3B(LC3B)and smooth muscle 22 alpha protein(SM22α)in vascular tissues at the aortic root were detected by immunofluorescence.The expressions of LC3B,Beclin-1 and SM22α in mouse vascular tissues were detected by Western Blot.qRT-PCR was used to detect the m RNA expressions of LC3B,Beclin-1 and SM22α.Results Compared with the blank group,HDL-C decreased,TC,TG and LDL-C increased(P<0.05).Oil red O,HE and Masson staining in the model group showed that plaque area at the aortic root was increased.The inner wall of blood vessels was thickened,which was accompanied by a lot of cholesterol crystals and infiltration of inflammatory cell,and the collagen fiber content was decreased.The results of immunofluorescence showed that the co-localization area of LC3B and SM22αwas significantly decreased(P<0.05).The protein and mRNA expressions of LC3B,Beclin-1 and SM22α were decreased(P<0.05).Compared with the model group,HDL-C was significantly increased,while TC,TG and LDL-C were significantly decreased(P<0.05)in positi

关 键 词：心脉康 软坚散结法 动脉粥样硬化 血管平滑肌细胞 自噬 ApoE^(-/-)小鼠 

分 类 号：R285.5[医药卫生—中药学]