摘要
目的观察高脂诱导的血管性ED家兔阴茎海绵体平滑肌(Corpus Cavernosum Smooth Muscle,CCSM)细胞形态、结构变化及高脂对CCSM细胞凋亡、增殖的影响。方法雄性新西兰家兔10只,随机分为模型组和对照组,每组各5只。模型组为采用经高脂、高胆固醇饮食结合球囊导管扩张双侧髂内动脉建立、鉴定的血管性ED模型;对照组喂以正常饮食。8周后切取阴茎CCSM组织观察并做体外细胞培养。经处理后分别在光镜、透射电镜下观察,流式细胞仪检测细胞凋亡,MTT比色法观察细胞增殖活性,RT-PCR法研究Caspase-3 mRNA的表达情况,免疫荧光法检测Caspase-3活性。结果与对照组相比,模型组光镜和电镜下CCSM细胞内含较多脂质囊泡,线粒体、内质网等细胞器出现损伤,凋亡小体明显增多。流式细胞仪检测证实模型组CCSM细胞凋亡率较对照组明显增高(1.44±0.20%Vs5.96±1.60%,P<0.01)。MTT比色法显示高脂对CCSM的增殖有明显抑制作用(模型组与对照组A值分别为:0.52±0.08Vs1.42±0.04,P<0.01)。模型组的Caspase-3 mRNA表达和活性值均显著高于对照组(0.74±0.00Vs0.65±0.00,P<0.01;活性值0.03±0.00Vs0.01±0.00,P<0.05)。结论高脂、高胆固醇饮食可严重影响CCSM结构,促进CCSM细胞凋亡并抑制其增殖。CCSM细胞凋亡/增生的失衡可能是高脂导致ED发生发展的重要病理环节。
Objective To study the ultrastructure changes, poliferation and apoptosis of Corpus Cavernous Smooth Muscle （CCSM） cells derived from hyperlipidemia-induced vasculogenic erectile dysfunction rabbits. Methods CCSM cells derived from experimental hyperlipidemia rabbit ED model＇s Smooth Muscle were cultured and its ultrastructural changes were observed by transmission electron microscope. Its proliferation was detected by MTT colorimetric assay. Then cell apoptosis was analyzed by flow cytometer. The expression of caspase-3 and activity were measured by RT-PCR and immunofluorescence. Results Significant pathological changes of ultrastructural were observed in the CCSM cells of ED rabbits such as condensation of cytoplasma, chromatin condensation, chromal cluping and margination and so on. The percentage of apoptotic CCSM cells in ED group was significantly higher than those in control groups （ 1.44±0.20% Vs 5.96±1.60%, P〈0.01 ）. and an inhibition effect on proliferation was found in ED group by MTT colorimetric assay （ 0.52±0.08 Vs 1.42±0.04, P〈0.01）. The mRNA expression and activity of Caspase-3 of CCSM cells in the ED group was significiantly higher than that in control group （mRNA exprssion： 0.74±0.00 Vs 0.65±0.00, P〈0.01; activity value： 0.03±0.00 Vs 0.01±0.00, P〈0.05）. Conclusion Hyperlipidimea can affect the the ultrastructure of the CCSM cell. It can also induce the CCSM cells＇ apoptosis and inhibit their proliferation, which might be one of the key mechanisms in ED induced by hyperlipidemia
出处
《中国男科学杂志》
CAS
CSCD
2008年第9期19-24,共6页
Chinese Journal of Andrology
关键词
高脂血症
勃起功能障碍
兔
细胞凋亡
细胞增殖
hyperlipidemia
erectile dysfunction
rabbits
apoptosis
cell proliferation
