不同浓度雌激素对大鼠膀胱颈平滑肌细胞增殖与凋亡的影响  

Effects of Different Concentrations of Estrogen on Proliferation and Apoptosis of Synthetic Bladder Neck Smooth Muscle Cells

作　　者：Wafi Walad Ali 李世文[1] 罗仪[1] 郑新民[1] 胡礼泉[1] 郑航[1] 胡万里[1] 

机构地区：[1]武汉大学中南医院泌尿外科,湖北武汉43007

出　　处：《武汉大学学报（医学版）》2008年第6期712-715,758,I0004,共6页Medical Journal of Wuhan University

基　　金：湖北省自然科学基金资助项目(编号:2007ABA285)

摘　　要：目的:探讨不同浓度17β-雌二醇(E2)对大鼠膀胱颈平滑肌细胞(BSMC)增殖与凋亡的影响。方法:无菌切取大鼠膀胱颈,应用酶消化法行原代细胞培养。取3-4代传代细胞,用α-平滑肌肌动蛋白抗体免疫组织化学法鉴定细胞成分,选取平滑肌成分大于85%者,分别加入不同浓度E2(0.1-100 nmol/L)。于处理72 h后收获细胞,应用流式细胞仪检测细胞周期、细胞凋亡及其相关蛋白CyclinD1,应用Western blotting法检测Bcl-2和Bax表达。结果:E2(0.1-10 nmol/L)促进BSMC从G1期向S过渡(G0/G1期比率显著低于对照组细胞,而S期和G2/M期比率显著高于对照组细胞,P<0.05),并伴随Cyclin D1蛋白表达显著增高。而大剂量E2(100 nmol/L)则使细胞凋亡率显著升高,并伴随Bax表达显著增加。结论:小剂量E2能够促进合成型BSMC增殖,其机制是影响Cyclin D1表达,加速G1期向S过渡;大剂量E2则通过增加Bax促进细胞凋亡。Objective： To investigate the effect of different concentrations of 17β-estradiol （E2） on proliferation and apoptosis of bladder neck smooth muscle cells（BSMCs） and the possible underlying mechanisms. Methods： The synthetic （subcultured for 3-4 passages） rat bladder neck smooth muscle cells were obtained by enzyme-digesting method and exposed to gradient concentrations （0.1-1 00 nmol/L） of E2for 72 h. The s-smooth muscle aetin antibody was used to identify the cells by immunohistochemistry. The progression of cell cycle, the apoptosis and the expressions of CyclinD1 protein were examined by flow cytometry. Bcl-2 and Bax proteins were detected via Western blotting method. Results： E2 （0.1-10 nmol/L） promoted BSMC proliferation by accelerating their cell cycle progression from G1 to S phases compared with that of control group（0 nmol/ L E2）. The rates of the BSMC at S and G2/M phase were significantly increased, while those at G0/G1 phase decreased. This was accompanied by an up-regulation of CyclinD1 expression. However, high dose of E2 （10 nmol/L and 100 nmol/L） increased the rate of apoptosis of the cells accompanied by a significant raise of Bax expression and the ratio of Bax/Bcl-2. Conclusion： The effect of E2 on subcultured （synthetic） BSMC is bilateral： it promotes BSMC proliferation by enhancing the expression of CyclinD1 which accelerates G1 to S phase transition, and on the other hand, it induces apoptosis of the cells by up-regulating the expression of Bax.

关 键 词：雌二醇 膀胱颈 平滑肌细胞 凋亡 细胞周期 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]