晚期糖基化终末产物诱导人卵巢颗粒细胞株COV434凋亡并上调促炎因子HMGB1自分泌

Advanced glycation end products induce apoptosis of human ovarian granulosa COV434 cells and up-regulate pro-inflammatory cytokine HMGB1 autocrine

目的:探讨晚期糖基化终末产物(advanced glycation end products,AGEs)是否诱导人卵巢颗粒细胞株COV434凋亡,并探究其可能的机制。方法:采用不同浓度AGEs牛血清白蛋白与人卵巢颗粒细胞株COV434共同孵育,流式细胞术观察细胞凋亡率,Western blot观察caspase-3和cleaved caspase-3的蛋白水平,ELISA检测细胞培养液上清中高迁移率族蛋白B1(high mobility group box 1 protein,HMGB1)的含量。结果:与对照组比较,100 mg/L AGEs组和200 mg/L AGEs组早、晚期凋亡率显著增加,各组caspase-3蛋白水平的差异无统计学显著性,但与对照组比较,100 mg/L AGEs组和200 mg/L AGEs组cleaved caspase-3的蛋白水平显著增加(P<0.05)。此外,与对照组比较,100 mg/L AGEs组和200 mg/L AGEs组细胞培养液上清中HMGB1促炎介质的水平显著上升(P<0.05)。结论:晚期糖基化终末产物诱导人卵巢颗粒细胞株COV434凋亡可能与促炎反应有关。

AIM:To study whether advanced glycation end products(AGEs)induce the apoptosis of human ovarian granulosa COV434 cells,and to explore the possible mechanism.METHODS:Human ovarian granulosa COV434 cells were treated with AGEs at different concentrations.Flow cytometry was used to observe the apoptotic rate.The protein levels of caspase-3 and cleaved caspase-3 were determined by Western blot.The release of high mobility group box 1 protein(HMGB1)in the culture supernatant was measured by ELISA.RESULTS:Compared with control group,early apoptotic rate and late apoptotic rate in 100 mg/L AGEs group and 200 mg/L AGEs group were significantly increased(P<0.05).No obvious difference of caspase-3 protein level in each group was observed,while the protein levels of cleaved caspase-3 in 100 mg/L AGEs group and 200 mg/L AGEs group were significantly increased compared with control group(P<0.05).In addition,compared with control group,pro-inflammatory factor HMGB1 in the culture medium in 100 mg/L AGEs group and 200 mg/L AGEs group was significantly increased.CONCLUSION:The apoptosis of human ovarian granulosa COV434 cells induced by AGEs may be related to pro-inflammatory reaction.