维生素C促进高糖环境下人脂肪干细胞的增殖及相关机制

发布时间：2019-01-26 21:15

【摘要】：背景:研究证实,维生素C除了具有抗氧化作用外,还具有促进细胞增殖的重要作用,但至今为止,有关维生素C对高糖环境下人脂肪干细胞的增殖作用及相关分子机制尚不明确。目的:研究维生素C促进高糖环境下人脂肪干细胞增殖的作用及相关分子机制。方法:取高塘环境中培养的第3代人脂肪干细胞,分别以0,100,150,200,250,300μmol/L维生素C处理,以低糖环境中培养的细胞为对照,MTT法筛选最佳维生素C干预浓度及处理时间,进行Western Blot检测。EdU检测:将高糖环境中培养的人脂肪干细胞分4组培养,空白对照组不进行任何干预,维生素C组加入最佳浓度维生素C,LY294002组加入PI3K/AKT信号通路抑制剂LY294002与最佳浓度维生素C,U0126组加入ERK/MAPK信号通路抑制剂U0126与最佳浓度维生素C,以低糖环境中培养的细胞为对照,干预48 h后,检测Ed U标记阳性细胞数。结果与结论:(1)CCK-8法检测:200μmol/L维生素C促进高糖环境下人脂肪干细胞的增殖作用最强,所以此为最佳干预浓度,最佳处理时间为48 h;(2)Western Blot检测:对照组、0μmol/L维生素C处理组、200μmol/L维生素C处理组p-AKT蛋白表达无差异,200μmol/L维生素C处理组p-ERK蛋白表达高于0μmol/L维生素C处理组(P0.01);(3)Ed U检测:维生素C组、LY294002组、对照组EdU标记阳性细胞数高于空白对照组(P0.01),维生素C组EdU标记阳性细胞数高于U0126组(P0.01);(4)结果表明:维生素C通过ERK/MAPK信号通路促进高糖环境下人脂肪干细胞的增殖。
[Abstract]:Background: studies have confirmed that vitamin C not only has antioxidant effects, but also plays an important role in promoting cell proliferation. The effect of vitamin C on the proliferation of human adipose stem cells in high glucose environment and its molecular mechanism are not clear. Aim: to study the effect of vitamin C on proliferation of human adipose stem cells in high glucose environment and its molecular mechanism. Methods: the third generation of human adipose stem cells cultured in Gaotang was treated with 0100150200250300 渭 mol/L vitamin C, and the cells cultured in low-sugar environment were used as control. The best intervention concentration and treatment time of vitamin C were screened by MTT method. Western Blot detection. EdU detection: human adipose stem cells cultured in high sugar environment were divided into 4 groups, the blank control group did not intervene, vitamin C group added the best concentration of vitamin C, LY294002 group added PI3K/AKT signaling pathway inhibitor LY294002 and best concentration of vitamin C, U0126 group added ERK/MAPK signal pathway inhibitor U0126 and optimum concentration of vitamin C, and cultured cells in low glucose environment were treated for 48 h. The number of Ed U labeled positive cells was detected. Results and conclusion: (1) CCK-8 assay showed that 200 渭 mol/L vitamin C had the strongest effect on the proliferation of human adipose stem cells in high glucose environment, so this was the best intervention concentration and the best treatment time was 48 hours. (2) Western Blot: there was no difference in p-AKT protein expression among control group, 0 渭 mol/L vitamin C treatment group and 200 渭 mol/L vitamin C treatment group. The expression of p-ERK protein in 200 渭 mol/L vitamin C treatment group was higher than that in 0 渭 mol/L vitamin C treatment group (P0.01). (3) Ed U detection: the number of EdU labeled positive cells in vitamin C group, LY294002 group and control group was higher than that in blank control group (P0.01), and the number of EdU labeled positive cells in vitamin C group was higher than that in U0126 group (P0.01). (4) the results showed that vitamin C promoted the proliferation of human adipose stem cells in high glucose environment through ERK/MAPK signaling pathway.
【作者单位】：广东医科大学附属医院整形外科研究所;
【基金】：2015年国家自然科学基金面上项目(81570260) 2014年广东省自然科学基金(2014A030313535) 2016广东省科技计划项目(2016A020214018) 2014湛江市财政资金科技专项经费资助项目(2014C01022) 2015广东医科大学博士基金(BJ201510) 2015年广东医科大学附属医院博士启动基金(BJ201505)~~
【分类号】：R587.2

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