CAJ | 학술논문

旨在研究microRNA126过表达时人正常肝细胞株葡萄糖代谢的改变。体外培养chang liver细胞系,优化转染条件确定核苷酸序列的转染浓度后,分别转染microRNA126的类似物、抑制物及相应的对照序列。实时荧光定量PCR测定microRNA126相对表达量以检测细胞转染效率。转染48 h后,加入100 nmol/L胰岛素和对应底物处理2 h,检测各组葡萄糖利用率、糖原合成量、糖异生以及糖酵解指标。结果显示实时荧光定量PCR测定microRNA126类似物组microRNA126相对表达量明显高于其他各组( P<0.05)。 microRNA126类似物组与其他各组相比,人正常肝细胞葡萄糖利用率下降,糖原合成量减少,糖异生增强,乳酸生成量减少,丙酮酸激酶活力下降(均P<0.05)。 microRNA126在肝细胞过表达时,可影响细胞的葡萄糖代谢,可能增加肝糖输出。
지재연구microRNA126과표체시인정상간세포주포도당대사적개변。체외배양chang liver세포계,우화전염조건학정핵감산서렬적전염농도후,분별전염microRNA126적유사물、억제물급상응적대조서렬。실시형광정량PCR측정microRNA126상대표체량이검측세포전염효솔。전염48 h후,가입100 nmol/L이도소화대응저물처리2 h,검측각조포도당이용솔、당원합성량、당이생이급당효해지표。결과현시실시형광정량PCR측정microRNA126유사물조microRNA126상대표체량명현고우기타각조( P<0.05)。 microRNA126유사물조여기타각조상비,인정상간세포포도당이용솔하강,당원합성량감소,당이생증강,유산생성량감소,병동산격매활력하강(균P<0.05)。 microRNA126재간세포과표체시,가영향세포적포도당대사,가능증가간당수출。
[Summary] To investigate the effect of microRNA126 on glucose metabolism in the normal liver cell lines. In vitro, the chang liver cell lines were cultured. Under the most effective transfection conditions ascertained above, microRNA126 mimic, microRNA126 inhibitor, and relative negative control were transfected into the cultured normal liver cells. And the transfection efficiency was tested by realtime fluorescent quantitative PCR. After 48 hours, the cells were stimulated with synthetic insulin ( 100 nmol/L ) and respective substrates for 2 hours. Then the glycogenesis, gluconeogenesis, and glycolysis in cells were measured. The level of microRNA126 of the microRNA126 mimic group was higher than the other groups, and the difference was statistically significant ( P<0. 05 ). MicroRNA126 mimic group significantly decreased glucose utilization, reduced glycogen synthesis, effectively increased the account of gluconeogenesis, reduced lactate production, and pyruvate kinase activity ( all P<0. 05). The over-expressing microRNA126 in hepatocytes may reverse the function of glucose metabolism, and enhance output of hepatic glucose.