高糖环境下西格列汀对大鼠心肌细胞纤维化过程中转录激活蛋白-1及骨膜蛋白表达的影响  

Effect of sitagliptin on the expression of transcriptional activator-1 and periostinin in the process of cardiomyocyte fibrosis under the environment of high sugar

作　　者：弓慧杰[1] 张光明 GONG Huijie;ZHANG Guangming(Department of Endocrinology and Nephrology,Xindu District People′s Hospital of Chengdu City,Chengdu 610500,Sichuan Province,China;Department of Pathophysiology,Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]成都市新都区人民医院内分泌/肾内科,四川成都610500 [2]西南医科大学病理生理学研究室,四川泸州646000

出　　处：《新乡医学院学报》2020年第9期830-834,共5页Journal of Xinxiang Medical University

摘　　要：目的探讨在高糖环境下西格列汀对大鼠心肌细胞纤维化过程中骨膜蛋白(POSTN)及转录激活蛋白-1表达(AP-1)的影响。方法将大鼠心肌细胞H9c2复苏后接种于含体积分数10%胎牛血清的改良型洛斯维1640培养基持续培养,并用无血清培养基继续培养使其同步化;将同步化的H9c2细胞分为对照组、高糖损伤组及西格列汀处理组,对照组H9c2细胞予以普通培养液,高糖损伤组H9c2细胞予以葡萄糖浓度为33.0 mmol·L^-1的培养基,西格列汀处理组H9c2细胞予以终浓度为33.0 mmol·L^-1葡萄糖+0.1μmol·L^-1西格列汀的培养基。采用细胞计数试剂盒-8检测细胞活力。酶联免疫吸附试验检测H9c2细胞上清液中Ⅲ型胶原蛋白(COLⅢ)表达水平;免疫组织化学法检测H9c2细胞中AP-1蛋白表达水平;反转录-聚合酶链反应检测H9c2细胞中POSTN和AP-1 mRNA表达水平。结果高糖损伤组和西格列汀处理组H9c2细胞增殖活力均显著高于对照组(P<0.05);西格列汀处理组H9c2细胞增殖活力显著低于高糖损伤组(P<0.05)。高糖损伤组和西格列汀处理组H9c2细胞中COLⅢ表达水平均显著高于对照组(P<0.05);西格列汀处理组H9c2细胞中COLⅢ表达水平均显著低于高糖损伤组(P<0.05)。高糖损伤组和西格列汀处理组H9c2细胞中AP-1表达水平显著高于对照组(P<0.05);西格列汀处理组H9c2细胞中AP-1表达水平显著低于高糖损伤组(P<0.05)。高糖损伤组和西格列汀处理组H9c2细胞中POSTN、AP-1 mRNA表达水平均显著高于对照组(P<0.05);西格列汀处理组H9c2细胞中POSTN、AP-1 mRNA表达水平均显著低于高糖损伤组(P<0.05)。结论体外高糖环境可促进大鼠心肌细胞的增殖活力,并使心肌细胞纤维化;西格列汀可通过抑制H9c2细胞中AP-1的过表达下调POSTN的表达,从而减缓纤维化进程,延缓心肌细胞损伤。Objective To investigate the effect of sitagliptin on the expression of periostin(POSTN)and transcriptional activator protein-1(AP-1)in the process of cardiomyocyte fibrosis under a high glucose environment.Methods Rat′s cardiomyocyte cells(H9c2 cells)were inoculated with 10%fetal bovine serum modified park roswell memorial institute 1640 medium for continuous culture after resuscitation,and then were further cultured in serum-free medium for synchronization;the synchronized cardiomyocytes were divided into the control group,the high glucose injury group and the sitagliptin treatment group.H9c2 cells in the control group was cultured in a normal medium,H9c2 cells in the high glucose injury group were cultured in a medium with concentration of 33.0 mmol·L^-1 glucose,and H9c2 cells in the sitagliptin treatment group were cultured in a medium with 33.0 mmol·L^-1 glucose and 0.1μmol·L^-1 sitagliptin.The cell activity was measured by cell counting kit-8 method.Enzyme linked immunosorbent assay was used to detect the expression level of typeⅢcollagen(COLⅢ)in the supernatant of H9c2 cells;immunohistochemistry was used to detect the expression of AP-1 in H9c2 cells;reverse transcription-polymerase chain reaction was used to detect the expression of POSTN and AP-1 mRNA in H9c2 cells.Results The proliferation activity of H9c2 cells in the high glucose injury group and the sitagliptin treatment group was significantly higher than that in the control group(P<0.05),while the proliferation activity of H9c2 cells in the sitagliptin treatment group was significantly lower than that in the high glucose injury group(P<0.05).The expression of COLⅢin the H9c2 cells in the high glucose injury group and the sitagliptin treatment group was significantly higher than that in the control group(P<0.05),and the expression of COLⅢof H9c2 cells in the sitagliptin treatment group was significantly lower than that in the high glucose injury group(P<0.05).The expression levels of POSTN and AP-1 mRNA in the high glucose injury group

关 键 词：西格列汀 大鼠心肌细胞 心肌细胞纤维化 骨膜蛋白 转录激活蛋白-1 

分 类 号：R542.23[医药卫生—心血管疾病]