ERβ基因敲除对小鼠阴茎海绵体eNOS-NO通路基因表达的影响

Effects of ERb Knockout on the Expression of eNOS-NO Pathway in Mouse Corpus Cavernosum Penis

目的:探讨雌激素受体β(ERβ)基因敲除对小鼠阴茎海绵体eNOS-NO通路基因表达的影响。方法:选取ERβ基因敲除(ERβKO)雄性小鼠和相应野生型雄性小鼠各12只,随机分为4组:正常对照组(A组)、ERβKO组(B组)、野生型+肿瘤坏死因子-α(TNF-α)处理组(C组)和ERβKO+TNF-α处理组(D组)。C组和D组每日给予TNF-α6μg/kg腹腔注射,A组及B组则以等量生理盐水替代,连续14 d。采用RT-PCR技术检测小鼠阴茎组织通路相关分子:eNOS、小窝蛋白-1(caveolin-1)、钙调蛋白(Cam)mRNA表达情况;Western blotting检测小鼠阴茎组织eNOS-NO通路相关分子蛋白表达水平的变化情况。结果:RT-PCR结果与Western blotting结果相符:与A组相比,B组eNOS、Cam表达减少,小窝蛋白-1表达上调(P<0.05);TNF-α给药后,与C组比较,D组的eNOS、Cam表达减少,小窝蛋白-1表达上调(P<0.05)。结论:ERβ基因敲除后,尤其在内皮损伤因素TNF-α作用下,eNOS-NO通路表达下调,提示ERβ对阴茎海绵窦内皮具有保护效应。

Objective： To investigate the effects of ERb knockout on the expression of eNOS-NO pathway in mouse corpus cavernosum penis.Methods： Twelve ERbKO male mice and 12 C57BL/6 male mice were randomly selected,and divided into 4 groups： normal wide control group（group A）,ERbKO group（group B）,wild-type ＋ TNF-a-treated group（group C） and ERbKO＋ TNF-a treated-group（group D）.TNF-a treated-groups were received intraperitoneal injection of TNF-a 6 mg/kg for 14 d,group A and group B were treated with saline instead.Then,the total mRNA was extracted from mouse penis,RT-PCR technique was used to detect the mRNA expression of relevant molecules in eNOS-NO pathway,such as eNOS,caveolin-1 and Cam;the total protein was extracted in mouse penile tissue to detect the protein expression of relevant molecules by Western blotting.Results： The results of RT-PCR consistent with the Western blotting： compared with group A,the expressions of eNOS,Cam were decreased,but the caveolin-1 was increased in group B,the difference was statistically significant（P〈0.05）;after the administration of TNF-a,compared with group C,the expressions of eNOS,Cam were decreased,but the expression of caveolin-1 was increased in group D,the difference was statistically significant（P〈0.05）.Conclusion： After the ERb knockout,especially treated with the endothelial injury factor of TNF-a,the expression of eNOS was reduced,and eNOS-NO pathway was down-regulated,suggesting that ERb has a protective effect on the penile cavernous sinus endothelium.