糖尿病性大鼠阴茎海绵体平滑肌细胞凋亡和增殖特征分析.PDFVIP

2012;32(2) 南方医科大学学报（J South Med Univ ） ·155 · 基础研究 糖尿病性大鼠阴茎海绵体平滑肌细胞凋亡和增殖特征分析 韦安阳，刘 洋，何书华，张 涛，吴子云，刘路浩 南方医科大学南方医院惠侨楼泌尿外科，广东 广州 510515 摘要：目的 了解糖尿病性大鼠阴茎海绵体平滑肌细胞（CCSM）凋亡和增殖特征，探讨其对糖尿病性大鼠CCSM 数量的影响。 方法利用链脲佐菌素建立糖尿病性大鼠模型。CCSM 原代培养，并进行免疫细胞化学染色鉴定。采用WST- 1检测细胞增殖 率，采用流式细胞仪检测细胞凋亡率，采用qRT-PCR 检测阴茎海绵体组织增殖细胞核抗原（PCNA）和凋亡蛋白caspase-3 mRNA 的表达。结果 造模后12周，糖尿病组和正常对照组大鼠体质量和随机血糖水平差异显著（P0.001）。糖尿病组大鼠阴 茎勃起功能较正常对照组大鼠差。原代培养所获细胞绝大部分为CCSM。糖尿病组CCSM 的增殖速度明显低于正常对照组 （P0.001）。在生长因子刺激下，糖尿病组CCSM增殖速率也慢于正常对照组（P0.05）。糖尿病组CCSM 凋亡细胞个数及凋亡 率均显著高于正常对照组（P0.001）。与正常对照组比较，糖尿病性大鼠阴茎海绵体组织中PCNA mRNA 表达下降，而 caspase-3 mRNA 表达增强，差异显著（P0.001）。结论 在长期糖尿病状态下，细胞凋亡增加与细胞增殖减慢并存，它们共同作 用导致CCSM数量明显减少。 关键词：勃起功能障碍；糖尿病；平滑肌细胞；细胞凋亡；增殖 中图分类号：R322.64; R587.1 文献标志码：A 文章编号：1673-4254（2012）02-0155-05 DOI: CNKI:44-1627/R1453.022 /kcms/detail/44.1627.R1453.022.html Apoptosis and proliferation of corpus cavernosum smooth muscle cells in diabetic rats WEI Anyang, LIU Yang, HE Shuhua, ZHANG Tao, WU Ziyun, LIU Luhao Department of Urology, Medical Center for Overseas Patients, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China Abstract: Objective To explore the characteristics of cell apoptosis and proliferation of corpus cavernosum smooth muscle (CCSM) cells in diabetic rats. Methods From a SD rat model of diabetes induced by a single dose of streptozotocin, CCSM cells were isolated for primary culture and identified using immunocytochemical assays for SMα-actin. The proliferation of CCSM cells was evaluated by WST-1 assay, and flow cytometry was used to detect the cells apoptosis. Real-time fluorescence quantitative RT-PCR (qRT-PCR) was used to analyze the relative expression of proliferation cell nucleus antigen (PCNA) and caspase-3 mRNA. Results The proliferation rate of the primarily cultured CCS