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作 者:许雪娟[1] 李焱[2] 刘珊英[2] 梁颖[2] 严励[2] 傅祖植[2]
机构地区:[1]中山大学附属佛山市第一人民医院内分泌科,528000 [2]中山大学附属第二医院内分泌科
出 处:《中华内分泌代谢杂志》2010年第5期402-405,共4页Chinese Journal of Endocrinology and Metabolism
基 金:教育部回国人员启动基金资助项目;广东省自然科学基金资助项目(6021329);国家自然科学基金资助项目(30671974)
摘 要:目的探讨抑制内源性微小RNA-375(miR-375)是否影响NIT-1胰岛β细胞的脂性凋亡。方法将NIT-1细胞分为4组:空白组(无脂质体)、空转组(脂质体)、阴性对照miRNA组(脂质体+阴性对照miRNA)、2'-O-me-375组(脂质体+2'-O-me-375,抑制内源性miR-375的作用)。转染72h后,各组细胞分别以500μmol/L棕榈酸孵育48h。以Hochest 33342染色和脱氧核糖核酸末端转移酶介导的缺口末端标记法(TUNEL)检测细胞凋亡,以Western印迹法检测miR-375靶基因肌营养素(myotrophin,V1)的表达。结果与其他3组比较,2’-O-me-375RNA组NIT-1脂性凋亡率最低(P〈0.01),同时V1表达水平最高(P〈0.01)。结论抑制内源性miR-375可降低胰岛β细胞的脂性凋亡。Objective To investigate the effect of miR-375 inhibitied by 2'-O-me-375 on lipoapoptosis of NIT-1 pancreatic β cells. Methods NIT-1 ceils were divided and treated according to the optimal condition:mock ( without lipofectamine), lipofectamine ( transfected only with lipofectamine), NC-miRNA ( transfected with negative control miRNA), and 2'-O-me-375 (transfected with 2'-O-me-375 )groups. 72 hours later, all ceils in each group were cultured with 500 μmol/L palmitate for 48 h. The percentage of apoptotic cells was detected by Hochest33342 staining and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL). The protein expression of myotrophin (V1), a target gene of miR-375, was detected by Western blotting. Results Compared to the other three groups, the cell apoptosis rate of 2'-O-me-375 group was the lowest (P〈0.01) ,along with the highest V1 expression level(P〈0.01 ). Conclusion Inhibition of miR-375 decreases pancreatic β-cell lipoapoptosis.
关 键 词:RNA干扰 Β细胞凋亡 微小RNA-375 肌营养素 2'-O-me-375
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