摘要
目的探讨血管紧张素Ⅱ1型受体(AT1R)在氧化三甲胺(TMAO)促动脉粥样硬化中的作用。方法用分子对接预测TMAO与AT1R可能的相互作用。将21只6周龄ApoE^(-/-)小鼠随机分为对照组、TMAO组、TMAO+替米沙坦组,每组7只。TMAO组在饲料中加1%胆碱复制高TMAO血症模型。TMAO+替米沙坦组采用替米沙坦10 mg/(kg·d)灌胃治疗。12周后采血,采用高效液相色谱串联质谱法测定血浆TMAO含量;油红O染色确定主动脉根部斑块面积;免疫组织化学法检测斑块内炎症因子单核细胞趋化蛋白-1(MCP-1)和白细胞介素-6(IL-6)的浸润;构建AT1R表达质粒,转染293T细胞,加入TMAO(200μmol/L)或TMAO(200μmol/L)+替米沙坦(1μmol/L)作用0、5、10、15、30和60 min,检测AT1R下游信号通路ERK、PKC磷酸化活化情况。结果分子对接预测到TMAO与AT1R之间的存在直接结合位点。3组小鼠主动脉根部斑块面积占比、斑块内MCP-1和IL-6表达比较,差异有统计学意义(P<0.05);TMAO组较对照组主动脉根部斑块面积占比增加(P<0.05),斑块内MCP-1和IL-6表达升高(P<0.05),而替米沙坦组较TMAO组主动脉根部斑块面积占比下降(P<0.05),斑块内MCP-1和IL-6表达降低(P<0.05)。3组主动脉组织的AT1R、p-ERK1/2、p-PKC蛋白相对表达量比较,差异有统计学意义(P<0.05);TMAO组较对照组升高(P<0.05),而替米沙坦组较TMAO组下降(P<0.05)。在转染AT1R质粒的293T细胞中,TMAO组(200μmol/L)不同时间点的p-ERK1/2、p-PKC蛋白相对表达量比较,差异有统计学意义(P<0.05);与0 min比较,作用15和30 min时p-ERK1/2蛋白相对表达量升高(P<0.05),作用10和15 min时p-PKC蛋白相对表达量升高(P<0.05)。而替米沙坦(1μmol/L)作用后,各个时间点的p-ERK1/2和p-PKC蛋白相对表达量比较,差异无统计学意义(P>0.05)。结论TMAO加重ApoE^(-/-)小鼠动脉粥样硬化机制可能包含了其对AT1R的作用。
Objective To explore the role of angiotensinⅡtype 1 receptor(AT1R)in the promotion of atherosclerosis by trimethylamine N-oxide(TMAO).Methods Molecular docking was used to predict the possible interaction between TMAO and AT1R.Twenty-one 6-week-old ApoE^(-/-)mice were randomly divided into control group,TMAO group,and TMAO+telmisartan group,with 7 mice in each group.The TMAO group was fed with 1%choline in the diet to replicate the high TMAO blood model.The TMAO+telmisartan group was treated with telmisartan 10 mg/(kg·d)by gavage.After 12 weeks,blood was collected,and plasma TMAO levels were determined by high-performance liquid chromatography-tandem mass spectrometry.Oil Red O staining was used to determine the plaque area at the aortic root.Immunohistochemistry was used to detect the infiltration of inflammatory factors monocyte chemoattractant protein-1(MCP-1)and interleukin-6(IL-6)in the plaque.AT1R expression plasmids were constructed,transfected into 293T cells,and treated with TMAO(200μmol/L)or TMAO(200μmol/L)+telmisartan(1μmol/L)for 0,5,10,15,30,and 60 min.The activation of the downstream signaling pathways ERK and PKC was detected.Results Molecular docking predicted a direct binding site between TMAO and AT1R.The proportion of plaque area at the aortic root,expression of MCP-1,and IL-6 in the plaque were statistically different among the three groups(P<0.05).The TMAO group showed an increase in the proportion of plaque area at the aortic root and an elevation of MCP-1 and IL-6 expression compared to the control group(P<0.05),while the telmisartan group showed a decrease compared to the TMAO group(P<0.05).The expression of AT1R,p-ERK1/2,and p-PKC in aortic tissues differed significantly among the three groups(P<0.05).The TMAO group increased significantly compared to the control group(P<0.05),while the telmisartan group decreased compared to the TMAO group(P<0.05).In 293T cells transfected with AT1R plasmids,the relative expression of p-ERK1/2 and p-PKC protein at different time points in the TMAO g
作者
李天翔
李素娟
郝翔宇
祝志波
郑梦
郭建强
Li Tian-xiang;Li Su-juan;Hao Xiang-yu;Zhu Zhi-bo;Zheng Meng;Guo Jian-qiang(Department of Cardiology,The Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia 010050,China;Department of Gastroenterology,The Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia 010050,China)
出处
《中国现代医学杂志》
CAS
2024年第1期34-43,共10页
China Journal of Modern Medicine
基金
内蒙古自治区自然科学基金(No:2023LHMS08065)。
