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中文摘要

《亚洲男性学杂志》 2010; 12 (5): 744-752

Tankyrase 1对老年性勃起功能障碍大鼠阴茎海绵体平滑肌细胞自噬的影响及其机制

比较老年性勃起功能障碍(ED)大鼠和勃起功能正常大鼠阴茎海绵体平肌细胞(CSMCs)中Tankyrase 1和自噬差别,探讨Tankyrase 1对老年性勃起功能障碍大鼠CSMCs中自噬和细胞增殖的影响及其机制。通过阴茎海绵体压力/平均动脉压(ICP/MAP)测定评价雄性Wistar大鼠阴茎勃起功能,从而挑选8月龄勃起功能正常和24月龄勃起功能障碍两组大鼠;CSMCs酶消化差速贴壁法体外分离原代培养、鉴定、纯度分析;比较两组CSMCs的Tankyrase 1和自噬水平。按LipofectamineTM方法将Tankyrase 1重组质粒转染大鼠CSMCs。以Tankyrase 1过表达前后的老年ED大鼠模型组CSMCs为对象,用MTT法分析细胞生长曲线,用流式细胞仪测定分析细胞周期,用透射电镜、MDC染色和GFP-LC3荧光定位显微镜检测CSMCs中自噬体,Western blot检测CSMCs中LC3-I/II、Beclin 1、p70S6K和p-p70S6K(Thr389)的表达。体外培养CSMCs经免疫荧光鉴定是平滑肌细胞,流式细胞仪分析其纯度为99.1%。与性功能正常大鼠组相比,老年ED大鼠组原代培养CSMCs中Tankyrase 1和自噬体含量明显降低(P < 0.01)。老年ED大鼠组Tankyrase 1过表达CSMCs的生长曲线增加(P < 0.05),细胞周期S期明显增加(P < 0.01),透射电镜、MDC染色和GFP-LC3显微镜下荧光定位检测自噬体含量明显增加(P < 0.01),自噬标记物LC3-I/II和自噬重要的正调节因子Beclin 1表达均明显增加(P < 0.01,P < 0.05),自噬负调节因子p70S6K表达未见明显差异,p-p70S6K(Thr389)表达均明显降低(P < 0.05)。本研究结果提示,老年性勃起功能障碍大鼠CSMCs中Tankyrase 1和自噬明显降低,Tankyrase 1过表达能明显增强CSMCs增殖,这可能与其明显增强自噬及其调节mTOR信号通路相关,相关机制有待于进一步研究。

关键词:衰老,自噬,阴茎海绵体,阴茎海绵体平滑肌细胞,勃起功能障碍,老年,tankyrase

 
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