白藜芦醇通过抑制miR-27b表达促进白色脂肪细胞的棕色化功能
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国家自然科学基金青年基金(81400840)


Resveratrol promotes the browning of white adipocytes via suppressing miR-27b expression
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    摘要:

    目的:探讨白藜芦醇对小鼠原代白色脂肪细胞棕色化功能的影响,研究白藜芦醇调控白色脂肪棕色化功能可能的分子机制。方法:从C57BL/5J小鼠的腹股沟处分离皮下前脂肪细胞,诱导分化后使用不同浓度的白藜芦醇(0~50 μmol/L)刺激48 h,运用RT-PCR检测棕色标志因子基因及相关的miR-27b表达情况。为了验证miR-27b的作用,在细胞内过表达miR-27b,同时予白藜芦醇刺激,运用RT-PCR和Western blot检测相关重要转录因子的基因及蛋白表达。结果:白藜芦醇促进了小鼠原代白色脂肪细胞棕色化功能基因的表达,抑制了原代白色脂肪细胞miR-27b表达,并呈剂量效应。过表达miR-27b显著抑制了白藜芦醇增强棕色化标志基因mRNA表达的作用。在蛋白水平上,过表达miR-27b同样抑制了白藜芦醇增强其靶基因PRDM16和棕色标记基因UCP1蛋白表达的作用。结论:白藜芦醇通过抑制miR-27b表达增强了白色脂肪细胞的棕色化功能,从而为临床有效改善机体能量代谢-防治代谢综合征提供了新策略。

    Abstract:

    Objective:To study the role of resveratrol in regulating the browning of mouse primary white adipocytes and find out the related molecular mechanisms. Methods:Primary subcutaneous preadipocytes were isolated from the groin of C57BL/5J mice and induced for differentiation. The cells were treated with different concentrations(0-50 -滋mol/L) of resveratrol to analyze the resveratrol′s effect on the browning of white adipocytes. Real-time PCR was performed to detect the mRNA expression of BAT-specific genes and miR-27. mimic-miR-27b was transfected into primary cells to increase miR-27b expression,then,resveratrol (50 μmol/L)was performed to stimulate these cells. We used real-time PCR and Western blot to measure mRNA and protein levels of BAT-specific genes in these cells. Results:Resveratrol promoted the expression of BAT-specific genes and suppressed miR-27b expression in a concentration-dependent manner in mouse white adipocytes. Overexpression of miR-27b significantly inhibited the resveratrol action on the mRNA expression of BAT-specific genes. At the protein level,overexpression of miR-27b also suppressed the resveratrol action on the PRDM16 and UCP1 protein levels in mouse white adipocytes. Conclusion:Our data showed that resveratrol promoted the browning of mouse white adipocytes via suppressing miR-27b expression. MiR-27b might provide a potential therapeutic target for the treatment of obesity and metabolic syndrome.

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孔小岑,刘炳丽,严仍娜,马建华.白藜芦醇通过抑制miR-27b表达促进白色脂肪细胞的棕色化功能[J].南京医科大学学报(自然科学版),2015,(11):1547-1551

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  • 收稿日期:2015-07-21
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  • 在线发布日期: 2015-11-30
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