载脂蛋白M对1–磷酸鞘氨醇受体–1表达的影响  

Effect of apolipoprotein M on the expression of sphingosine-1-phosphate receptor 1

作　　者：王敏[1] 罗光华[2] 张友浦 黄波[1] 于洋[2] 刘宏[1] 徐宁[3] 张晓膺[1] Wang Min;Luo Guanghua;Zhang Youpu;Huang Bo;Yu Yang;Liu Hong;Xu Ning;Zhang Xiaoying(Department of Cardiothoracic Surgery,the Third Affiliated Hospital of Soochow University,Changzhou 213000,China;Comprehensive Laboratory,the Third Affiliated Hospital of Soochow University,Changzhou 213000,China;Department of clinical chemistry,Medical Laboratory of Lund University,Sweden)

机构地区：[1]苏州大学附属第三医院心胸外科,江苏常州213000 [2]苏州大学附属第三医院综合实验室,江苏常州213000 [3]瑞典隆德大学医学实验室临床化学系

出　　处：《中华肥胖与代谢病电子杂志》2017年第2期91-96,共6页Chinese Journal Of Obesity and Metabolic Diseases：Electronic Edition

基　　金：国家自然科学基金(81370372,81201352);江苏省自然科学基金(BK2012154,BK20130244,BK20151179);江苏省“333工程”科研资助项目(BRA2013062);常州市国际科技合作项目(CZ20120017);常州市卫生人才培养工程资助(2016ZCJ002)

摘　　要：目的探讨载脂蛋白M(apoM)对1–磷酸鞘氨醇受体–1(S1P1)表达的影响。方法体外实验:采用载有人apoM基因序列(实验组)和不携带apoM基因序列(对照组)的慢病毒分别感染人脐静脉内皮融合细胞(EA.hy926细胞),获得稳定表达的细胞株后提取细胞总RNA和总蛋白,采用RT-PCR技术检测apoM mRNA的过表达情况,并采用RT-PCR和Western Blot法检测S1P1受体的表达情况。体内实验:随机选取8~10周,体质量约25 g的健康apoM基因野生型(apoM^(+/+),实验组)和apoM基因敲除型(apoM^(–/–),对照组)C57BL/6小鼠各8只,处死小鼠后取其主动脉提取肝脏RNA,逆转录成cDNA后采用RTPCR法检测apoM和S1P1 mRNA的表达情况。分别比较体内和体外实验中实验组和对照组apoM mRNA和S1P1蛋白及mRNA的表达情况。结果 (1)实验组EA.hy926细胞的S1P1 mRNA和蛋白表达水平均明显高于对照组细胞(P<0.05);(2)实验组apoM^(+/+)小鼠主动脉S1P1 mRNA和蛋白表达水平也明显高于对照组apoM^(–/–)小鼠(P<0.05)。结论 ApoM能够增加体内和体外S1P1 mRNA及蛋白表达水平。Objective To investigate the effect of apolipoprotein M(apoM)on the expression ofsphingosine-1-phosphate receptor1(S1P1).Methods In the vitro experiment,lentiviruses with apoM gene(the experimental group)and without apoM gene(the control group)were used to infect the endothelialcells of human umbilical vein(EA.hy926cells).Then the total mRNAs and proteins were obtained fromthe cell lines with stable expression,and RT-PCR was used to detect the expression level of apoM,RT-PCRand Western Blot were used to detect the expression level of S1P1receptor.In the vivo experiment,8~10week-old and25-gram-weight healthy wild-type apoM gene(apoM+/+,the experimental group)and apoMknockout(apoM-/-,the control group)C57BL/6mice were selected,with8mice in each group.After theywere sacrificed,the liver mRNAs were extracted from the aortas and reversely transcribed to cDNAs.ThenRT-PCR was used to detect the expression level of apoM and S1P1mRNA.The expressions levels of apoMmRNA,S1P1mRNA and protein were compared between the experimental and control group in the vitro andvivo experiments.Results(1)The expression levels of S1P1mRNA and protein of the EA.hy926cells inthe experimental group were significantly higher than those of the EA.hy926cells in the control group(P<0.05);(2)the expression levels of S1P1mRNA and protein in the aorta of apoM+/+mice were significantly higher thanthose of the apoM-/-mice(P<0.05).Conclusion ApoM can increase the expression level of S1P1mRNAand protein in vivo and vitro.

关 键 词：载脂蛋白M 1–磷酸鞘氨醇受体–1 信使RNA 受体表达 

分 类 号：R543.5[医药卫生—心血管疾病]