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目的：探究microRNA-195 对棕榈酸诱导肝细胞L02 葡萄糖摄取障碍中的影响及其作用机制。方法：体外培养人肝细胞 L02，用不同浓度棕榈酸(0.6 mmol/L，1 mmol/L)诱导人肝细胞L02(10 h)，显微镜观察其形态学变化。应用葡萄糖氧化酶法检测细胞培养液中剩余的葡萄糖含量，Real Time-PCR 检测棕榈酸诱导肝细胞L02 葡萄糖摄取障碍后microRNA-195 (miR-195)表达变化，Real Time-PCR检测转染miR-195 mimics 及miR-195 inhibitor后miR-195 的表达水平，激光共聚焦检测转染效率，miR-195 过表达或低表达后葡萄糖摄取的变化。结果：高浓度棕榈酸可诱导肝细胞L02葡萄糖摄取障碍，培养液上清糖浓度升高约2.5 mmol/L， miR-195 表达量明显升高；转染miR-195 mimic 后miR-195表达升高约200 倍，转染miR-195 inhibitor 后miR-195 表达降低约40 倍；过表达miR-195 后细胞培养液上清中葡萄糖量升高2.5 mmol/L，即葡萄糖摄取降低；低表达miR-195 后上清液中葡萄糖量降低约1 mmol/L，即葡萄糖摄取作用加强。结论：miR-195 有可能参与调控肝细胞L02 葡萄糖摄取障碍。

英文摘要:

Objective:To study the influence and Novel Function mechanismof microRNA-195 in the process of Glucose Uptake Disorder Induced By PA in Human Liver Cell L02.Methods:Human Liver Cell L02 was cultured in vitro and treated with different concentrations of PA (0.6 mmol/L, 1 mmol/L) for 10 h. The morphological changes of Human Liver Cell L02 were detected by microscope. GOD-POD detect the rest glucose content in cell culture, the expression levels of miR-195 was analyzed by Real time-PCR after induced by PA. Measured fluorescence intensity by Laser confocal scanning microscopy, and the change of glucose uptake by up-regulated and down-regulated of miR-195.Results:High concentration PA could induced Liver Cell L02 Glucose Uptake Disorder, the glucose content increased 2.5 mmol/L, and miR-195 over-expressioned. After transfection of miR-195 mimics, the expression of miR-195 up-regulated, and transfection of miR-195 inhibitor the expression of miR-195 down regulated. The concentration of glucose in cell culture was increased 2.5 mmol/L when miR-195 over-expressioned, that is to say glucose uptake reduced. The concentration of glucose decreased 1 mmol/L in cell culture decreased when miR-195 low-expressioned.Conclusion:miR-195 is very likely to regulate the disorder of glucose uptaken in.

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