史美娜 刘寒强 张磊 杨瑞华 王枫.microRNA-195 在棕榈酸诱导肝细胞L02 葡萄糖摄取障碍中的作用[J].,2016,16(11):2041-2044 |
microRNA-195 在棕榈酸诱导肝细胞L02 葡萄糖摄取障碍中的作用 |
A Novel Function of microRNA-195 in Human Liver Cell L02 GlucoseUptake Disorder Induced by PA |
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DOI: |
中文关键词: microRNA-195(miR-195) 棕榈酸 葡萄糖摄取 转染 |
英文关键词: MicroRNA-195(miR-195) PA Glucose uptake Transfection |
基金项目:国家自然科学基金面上项目(81072303) |
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中文摘要: |
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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