脂肪酸结合蛋白4在同型半胱氨酸致肝细胞脂质沉积中的作用  

Role of Fatty Acid Binding Protein 4 in Homocysteine-induced Lipid Deposition in Hepatocytes

作　　者：邓梅[1,2,3] 李桂忠 谢琳[1,2,3] 张辉 郭伟[1,2,3] 丁宁 刘圆[1,2,3] 孙卓成 马胜超[1,2,3] 张鸣号 杨晓玲 姜怡邓 DENG Mei;LI Guizhong;XIE Lin;ZHANG Hui;GUO Wei;DING Ning;LIU Yuan;SUN Zhuocheng;MA Shengchao;ZHANG Minghao;YANG Xiaoling;JIANG Yideng(Department of Preclinical Medicine College,Ningxia Medical University,Yinchuan750004,China;Ningxia Key Laboratory of Vascular Injury and Repair Research,Yinchuan750004,China;Key Laboratory of Metabolic Cardiovascular Diseases Research of National Health Commission,Yinchuan750004,China)

机构地区：[1]宁夏医科大学基础医学院,银川750004 [2]宁夏血管损伤与修复研究重点实验室,银川750004 [3]国家卫健委代谢性心血管疾病研究重点实验室,银川750004

出　　处：《宁夏医科大学学报》2020年第6期541-546,共6页Journal of Ningxia Medical University

基　　金：国家自然科学基金(81870225);宁夏回族自治区重点研发计划重大项目(2018BEG02004);宁夏高等学校一流学科建设(宁夏医科大学西部一流建设学科基础医学)项目(NXYLXK2017B07);大学生创新创业项目(NXCY2016126)。

摘　　要：目的探讨脂肪酸结合蛋白4(fatty acid binding protein 4,FABP4)对同型半胱氨酸(homocysteine,Hcy)致肝细胞脂质沉积的促进作用。方法胱硫醚β-合成酶(cystathionineβ-synthase,CBS)基因缺陷杂合体(CBS+/-)小鼠CBS基因正常(CBS+/+)小鼠给予高蛋氨酸饮食12周,设CBS+/+Met组(CBS+/+小鼠高蛋氨酸饮食)和CBS+/-Met组(CBS+/-小鼠高蛋氨酸饮食);体外培养人正常肝细胞HL-7702并给予100μmol·L^-1Hcy干预,FABP4过表达腺病毒转染肝细胞。免疫荧光双染观察小鼠肝脏组织中脂质分化相关蛋白(adipose differentiation-related protein,ADFP)表达;实时定量PCR(RT-qPCR)和Western blot分别检测小鼠肝脏和人肝细胞HL-7702中FABP4的表达;肝细胞HL-7702油红O染色观察肝细胞内脂质状况。结果 CBS+/+Met和CBS+/-Met组小鼠的肝脏组织中肝细胞表面标记物Cytokeratin18呈现绿色荧光,与CBS+/+Met组相比,CBS+/-Met组小鼠肝脏中ADFP表达明显增多(P<0.01);肝脏中FABP4的mRNA(P<0.01)和蛋白(P<0.05)表达均升高。Hcy干预肝细胞HL-7702后胞质中红色脂质颗粒明显增多(P<0.05),FABP4的表达趋势与动物水平一致,均明显升高(P<0.01);在有或无Hcy干预肝细胞的情况下转染FABP4过表达腺病毒后肝细胞胞质内红色脂质颗粒均明显增多(P均<0.01)。结论 FABP4的上调可能促进Hcy诱导的肝细胞脂质沉积。Objective To investigate the effect of fatty acid binding protein 4(FABP4)on lipid deposition in hepatocytes induced by homocysteine(Hcy). Methods Cystathionine beta-synthase(CBS) gene deficient heterozygote(CBS+/-)mice with normal CBS gene(CBS+/+)were fed with high methionine diet for 12 weeks. The mice were divided into two groups:CBS+/+Met group(CBS+/+high methionine diet)and CBS+/-Met group(CBS+/-mice high methionine diet). Human normal hepatocytes HL-7702 were cultured in vitro and treated with100 μmol·L^-1 Hcy. FABP4 overexpression adenovirus was transfected into hepatocytes. The expression of lipid differentiation associated protein(ADFP)in mouse liver was observed by immunofluorescence double staining;the expression of FABP4 in mouse liver and human hepatocyte HL-7702 was detected by real-time quantitative PCR(RT-qPCR)and Western blot,and the lipid status of hepatocyte was observed by HL-7702 oil red O staining. Results Cytokeratin18,a hepatocyte surface marker,showed green fluorescence in CBS+/+Met and CBS+/-Met groups. Compared with CBS+/+Met group,the expression of ADFP in CBS+/-Met group was significantly increased(P<0.01),and the m RNA(P<0.01)and protein(P<0.05)expression of FABP4 in liver were increased.The number of red lipid particles in cytoplasm of hepatocytes treated with Hcy was significantly increased(P<0.05),and the expression trend of FABP4 was the same as that of animals(P<0.01),and the number of red lipid particles in cytoplasm of hepatocytes transfected with adenovirus overexpressed by FABP4 was significantly increased with or without Hcy intervention(P all<0.01). Conclusion The up-regulation of FABP4 may promote the lipid deposition of hepatocytes induced by Hcy.

关 键 词：脂肪酸结合蛋白4 同型半胱氨酸 肝细胞 脂质沉积 

分 类 号：R575.5[医药卫生—消化系统]