二甲双胍对糖基化终末产物诱导的成纤维细胞凋亡及相关蛋白caspase-3、Bax及Bcl-2表达的影响  

Effects of metformin on apoptosis induced by advanced glycation end-products and expressions of caspase-3, Bax and Bcl-2 in human dermal fibroblasts in vitro

作　　者：庞若宇 关美萍[1] 郑宗基[1] 薛耀明[1] 

机构地区：[1]南方医科大学南方医院内分泌代谢科,广东广州510515

出　　处：《南方医科大学学报》2015年第6期898-902,共5页Journal of Southern Medical University

基　　金：广东省级产业技术研究与开发专项(2060403)

摘　　要：目的探讨晚期糖基化终末产物(AGEs)对原代人皮肤成纤维细胞凋亡的影响,以及二甲双胍(Metformin)对原代皮肤成纤维细胞凋亡是否存在保护作用。方法取对数生长期的皮肤成纤维细胞,分为空白组,BSA对照组(300μg/m L),AGEs刺激组(100、200、300μg/m L),药物组(AGEs 300μg/m L+Metformin 1 mmol/L),采用CCK-8检测24、48、72 h细胞凋亡情况;Western Blot检测细胞培养72 h后凋亡相关蛋白caspase-3、Bax、Bcl-2表达情况。结果 CCK-8结果显示AGEs诱导皮肤成纤维细胞凋亡呈浓度及时间依赖性,AGEs 300μg/m L诱导72 h后可见细胞凋亡明显增多(0.72±0.02 vs 1±0.04,P<0.05),加入二甲双胍后可对成纤维细胞起一定保护作用(0.98±0.02 vs 0.72±0.02,P<0.05)。Western Blot显示AGEs 300μg/m L刺激成纤维细胞72 h后,caspase-3、Bax蛋白表达增加(P<0.05),而Bcl-2蛋白表达下降(P<0.05),Bcl-2/Bax比值下降(P<0.05),二甲双胍作用后caspase-3、Bax蛋白表达水平较刺激组明显下降(P<0.05),而Bcl-2及Bcl-2/Bax比值则明显上升(P<0.05)。结论 AGEs可诱导人皮肤成纤维细胞凋亡增加,二甲双胍可以通过调控caspase-3、Bax及Bcl-2表达,从而起到抗凋亡作用。Objective To investigate the effect of metformin in protecting against advanced glycation end products（AGEs）-induced apoptosis in human primary dermal fibroblasts. Methods Fibroblasts were exposed to 100, 200, or 300 μg/m L AGEs,300 μg/m L bovine serum albumin（BSA）, or 300 μg/m L AGEs and 1 mmol/L metformin for 24, 48, or 72 h. The exposed cells were examined for cell apoptosis using a cell counting kit. The expressions of caspase-3, Bax and Bcl-2 protein in the fibroblasts treated for 72 h were detected with Western blotting. Results AGEs exposures caused significant dose- and time- dependent apoptosis in the fibroblasts. A 72- h exposure to 300 μg/m L AGEs resulted in obviously increased apoptosis of the fibroblasts compared to the control group（0.72±0.02 vs 1±0.04, P〈0.05）, and metformin significantly decreased AGEs-induced apoptosis（0.98± 0.02 vs 0.72 ±0.02, P〈0.05）. The expressions of caspase-3 and Bax protein were significantly increased（P〈0.05） and Bcl-2protein expression was decreased（P〈0.05） with a lowered Bcl- 2/Bax ratio in AGEs- treated fibroblasts（P〈0.05）, and such changes were significantly reversed by metformin treatment（P〈0.05）. Conclusion Metformin can antagonize AGEs- induced apoptosis in human dermal fibroblasts by regulating the expressions of caspase-3, Bax and Bcl-2.

关 键 词：糖尿病足溃疡 晚期糖基化终末产物 皮肤成纤维细胞 细胞凋亡 二甲双胍 

分 类 号：R96[医药卫生—药理学]