茱萸丸调控TMA/FMO3/TMAO通路促进胆固醇逆向转运抗动脉粥样硬化  

Zhuyu Pills(茱萸丸)Regulate TMA/FMO3/TMAO Pathway and Promote Cholesterol Reverse Transport and Anti-Atherosclerosis

作　　者：宋玮 梁清芝 张钟艺 沈涛[1] SONG Wei;LIANG Qingzhi;ZHANG Zhongyi;SHEN Tao(Chengdu University of Traditional Chinese Medicine,Chengdu 610075,Sichuan,China)

机构地区：[1]成都中医药大学,四川成都610075

出　　处：《中华中医药学刊》2024年第9期124-129,I0020-I0022,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81973743);四川省科技厅科技创新创业人才和苗子工程项目(20MZGC0241);四川省科技厅重点研发项目(2022YFS0381)。

摘　　要：目的旨在研究茱萸丸对动脉粥样硬化(atherosclerosis,AS)形成的影响及其相关机制。方法采用高脂饲料喂养雄性载脂蛋白E基因敲除(ApoE^(-/-))小鼠诱导动脉粥样硬化模型,造模周期为12周。造模成功的47只ApoE^(-/-)小鼠随机分成5组,即模型组10只,茱萸丸低、中、高剂量组各9只,阿托伐他汀钙组10只,以同周龄雄性C57BL/6J小鼠10只作为空白组。空白组与模型组予等体积无菌蒸馏水灌胃,茱萸丸低、中、高剂量组分别给予130.54、261.08、522.16 mg·kg^(-1)灌胃,阿托伐他汀钙组10.40 mg·kg^(-1)灌胃,每日1次,各组小鼠共灌胃12周。给药结束后,采用苏木精-伊红(HE)染色观测主动脉及肝脏病理变化;酶联免疫吸附测定法(ELISA)检测血清中氧化型低密度脂蛋白(ox-LDL)、单核细胞趋化蛋-1(MCP-1)、血管细胞黏附分-1(VCAM-1)、细胞间黏附分子-1(ICAM-1)水平;生化法检测肝脏总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)水平;超高效液相色谱-质谱联用技术(UHPLC-MS/MS)检测血浆三甲胺(TMA)、氧化三甲胺(TMAO)含量;免疫荧光法检测小鼠肝脏中二甲基苯胺单加氧酶3(FMO3)蛋白的表达水平;实时荧光定量聚合酶链式反应(real-time PCR)检测肝脏FMO3、腺苷三磷酸结合盒转运蛋白A1(ABCA1)、B族Ⅰ型清道夫受体(SR-B1)、ATP结合盒转运子G5抗体(ABCG5)、三磷酸腺苷结合盒转运体G8(ABCG8)和细胞色素P4507A1(CYP7A1)mRNA表达水平。结果与空白组比较,模型组小鼠HE染色可见主动脉内膜大面积增厚,肝脏脂肪变性及炎性浸润严重;血清中ox-LDL、MCP-1、VCAM-1、ICAM-1升高;肝脏TC、TG、LDL-C升高;血浆中TMA、TMAO水平升高;以及肝脏中FMO3 mRNA与蛋白表达水平升高,ABCA1、SR-B1、ABCG5、ABCG8、CYP7A1 mRNA表达水平升高(P<0.01)。与模型组比较,茱萸丸低、中、高剂量组及阿托伐他汀钙组HE染色随着茱萸丸浓度的增加,斑块富集区域逐渐缩小,肝脏脂肪变性及Objective This article aims to investigate the effect of Zhuyu Pills(茱萸丸)on atherosclerosis and the underlying mechanism.The mouse model of atherosclerosis was induced by a high-fat diet,and the total modeling period was 12 weeks.Methods A total of 47 ApoE-/-mice successfully modeled were randomized into 5 groups,including 10 in the model group,9 in each of low,medium and high dose(130.54,261.08 and 522.16 mg·kg^(-1),respectively)Zhuyu Pills groups,and 10 in the ator-vastatin calcium(10.40 mg·kg^(-1))group.In addition,10 C57BL/6J mice were included as the normal group.The mice in the normal group and model group were administrated with an equal volume of sterile distilled water,and those in the other groups were administrated with the corresponding agents by gavage once a day for 12 weeks.The pathological changes of aorta and liver were observed by hematoxylin-eosin(HE)staining after administration.The levels of oxidized low-density lipoprotein(ox-LDL),monocyte chemotactic egg-1(MCP-1),vascular cell adhesion-1(VCAM-1)and intercellular adhesion molecule-1(ICAM-1)in serum were detected by enzyme-linked immunosorbent assay(ELISA).The levels of total cholesterol(TC),tri-glyceride(TG)and low density lipoprotein cholesterol(LDL-C)in liver were detected by biochemical method.Ultra high per-formance liquid chromatography-mass spectrometry(UHPLC-MS/MS)was used to detect the plasma trimethylamine(TMA)and trimethylamine oxide(TMAO).The expression level of flavin containing monooxygenase 3(FMO3)protein in mouse liver was detected by immunofluorescence.Real-time fluorescence quantitative polymerase chain reaction(real-time PCR)was em-ployed to measure the mRNA levels of FMO3,adenosine triphosphate binding cassette transporter A1(ABCA1),scavenger recep-tor B1(SR-B1),ATP binding cassette transporter G5(ABCG5),adenosine triphosphate-binding cassette transporter G8(AB-CG8)and cytochrome P4507A1(CYP7A1)in the liver tissues.Results Compared with the control group,the model group showed large area thickening of the aortic in

关 键 词：茱萸丸 动脉粥样硬化 胆固醇逆向转运 TMA/FMO3/TMAO脂代谢通路 

分 类 号：R289.5[医药卫生—方剂学]