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目的研究雄激素对eNOS和PDE5蛋白在同一动物模型中表达的影响。方法用化学发光法测定新西兰白兔睾酮基线水平和去势后各个时间点的睾酮水平;用Western blot和免疫组化方法分析各组兔阴茎海绵体eNOS和PDE5蛋白的表达。结果假手术对照组(C组)兔血清睾酮水平和基线值没有显著性差异;去势组(Ox组)睾酮水平在2、4、8周时下降为基线值的17%、14%和13%;睾酮补充治疗组(OxT组)睾酮水平在不同剂量睾酮补充之后有所回升。C组eNOS和PDE5蛋白的表达不随时间的推移而变化;Ox组中,eNOS的表达速度、程度和时间推移的关系不明显,但是,PDE5蛋白的表达水平随去势时间的延长而降低;OxT组中eNOS蛋白表达水平没有明显差异,PDE5蛋白的表达水平在补充睾酮前后差异明显。结论在去势兔模型中,eNOS和PDE5蛋白表达不均衡,致使cGMP的生成和降解不能保持动态平衡,提示运用雄激素维持cGMP的动态平衡是ED治疗机理上的新视角。
Objective To investigate the effect of androgen on the expression of eNOS and PDE5 in the same animal model. Methods The levels of testosterone in New Zealand white rabbits at baseline and at various time points after castration were determined by chemiluminescence method. The expressions of eNOS and PDE5 protein in the penis of each group were detected by Western blot and immunohistochemistry. Results There was no significant difference between baseline and serum testosterone levels in the sham operation control group (C group). Testosterone levels in the castrated group (Ox group) dropped to 17%, 14% and 13 of the baseline at 2, 4 and 8 weeks %; Testosterone supplementation group (OxT group) testosterone levels increased after different doses of testosterone supplementation. The expression of eNOS and PDE5 in group C did not change with the passage of time. In Ox group, the expression of eNOS was not related to the time, but the expression level of PDE5 protein decreased with the increase of time of ovariectomy. There was no significant difference in eNOS protein expression between groups. The expression of PDE5 protein was significantly different between before and after testosterone supplementation. Conclusions The expression of eNOS and PDE5 protein is not balanced in castrated rabbit model, which leads to that the generation and degradation of cGMP can not maintain the homeostasis. It is suggested that the use of androgen to maintain the homeostasis of cGMP is a new perspective on the mechanism of ED treatment.