雌二醇、丙酸睾酮对泪腺细胞MMP-9表达及细胞凋亡的作用机制

Effects of estradiol and testosterone propionate on MMP-9 expression and apoptosis of lacrimal gland cells

目的:研究雌二醇(estradiol,E2)和丙酸睾酮(testosterone propionate,TP)对兔泪腺上皮细胞凋亡及对MMP-9蛋白表达的影响,探讨雌二醇及丙酸睾酮对干眼的作用机制。方法:体外培养兔泪腺上皮细胞,分别给予雌二醇及丙酸睾酮处理,双氧水(H_2O_2)诱导细胞凋亡,流式细胞仪检测细胞凋亡和免疫细胞化学法检测细胞MMP-9蛋白表达。实验分4组:凋亡对照组(AC组)、雌二醇组(E2组)、丙酸睾酮组(TP组)和空白对照组(BC组)。AC组只给予H_2O_2诱导凋亡,E2组给予1×10^(-5) mol/L雌二醇处理,TP组给予丙酸睾酮处理,BC组未加药物干预及未加H_2O_2诱导凋亡。结果:经H_2O_2诱导细胞凋亡后,泪腺上皮细胞早期凋亡率和MMP-9蛋白表达累积光密度值与BC组相比,AC组、E2组及TP组均显著增加;同比AC组,E2组及TP组细胞凋亡率和MMP-9蛋白表达量降低;E2组凋亡率及蛋白表达量比TP组明显降低;其差异均具有统计学意义(P<0.01)。结论:雌二醇和丙酸睾酮对H_2O_2诱导兔泪腺上皮细胞凋亡有一定的抑制作用,同时泪腺上皮细胞中MMP-9表达含量也降低,提示雌二醇和丙酸睾酮抑制兔泪腺上皮细胞凋亡的作用机制可能与MMP-9有关。

Objective:To investigate the effects of estradiol(E2)and testosterone propionate(TP)on apoptosis and matrix metalloproteinase-9(MMP-9)expression in rabbit lacrimal gland epithelial cells.Methods:The rabbit lacrimal gland epithelial cells were cultured in vitro.H2O2 was used to induce apoptosis in cultured lacrimal gland epithelial cells and then treated with E2 and TP respectively.Cell apoptosis was detected by flow cytometer(FCM)and MMP-9 expression was evaluated by immunocytochemistry.There were four groups:apoptosis control group(AC),estradiol group(E2),testosterone propionate group(TP)and blank control group(BC)respectively.The cells of group AC were administrated with H2O2 only,group E2 and group TP with 1×10?5 mol/L E2 and TP respectively and group BC in treated.Statistical analysis were performed with one-way analysis of variance(ANOVA test)using SPSS 16.0,P<0.05 was considered statistically significant.Results:Compared with group BC,the early cells apoptosis rate and integrated optical density of MMP-9 expression of lacrimal gland in group AC,E2 and TP increased significantly after the cells were induced by H2O2;Compared with group AC,group E2 and TP reduced;group E2 were significantly lower than group TP;the differences were all statistically significant(P<0.01).Conclusion:E2 and TP had a certain inhibitory effect on rabbit lacrimal gland epithelial cells apoptosis induced by H2O2.Meanwhile,MMP-9 expression of the lacrimal gland cells was decreased.These results indicated that E2 and TP on the apoptosis inhibitory mechanism of lacrimal gland cells may be related with MMP-9.