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目的:探讨ERβ基因过表达对ERβKO小鼠阴茎血管内皮的影响及相关分子机制。方法:选取ERβKO雄性小鼠12只,随机分为两组:ERβKO+TNFα+pAdxsi-ERβ组和ERβKO+TNFα+空病毒组。ERβKO+TNFα+pAdxsi-ERβ组给予ERβ基因重组腺病毒转染处理14 d,ERβKO+TNFα+空病毒组转染空病毒作对照,同时两组均给予TNFα6μg/(kg·d)腹腔注射,连续14 d。采用APO法观察小鼠的阴茎勃起功能;免疫组化检测内皮标志物CD34、vWF的变化情况;RT-PCR、Western印迹和免疫组化检测eNOS-NO通路相关分子表达情况。结果:与ERβKO+TNFα+空病毒组相比,ERβKO+TNFα+pAdxsi-ERβ组小鼠的变化情况如下:①勃起次数增多(2.17±0.41 vs 0.50±0.55,P<0.05),勃起潜伏期缩短[(24.0±1.27)min vs(28.83±1.33)min,P<0.05];②内皮标志物CD34、vWF表达更丰富[(1.50±0.55;1.33±0.52)vs(0.67±0.52;0.50±0.55),P<0.05];③eNOS、Cam表达减少,小窝蛋白-1表达上调(P均<0.05),RT-PCR与Western印迹结果相符合。结论:ERβ基因对阴茎血管内皮具有保护作用,eNOS-NO通路是其发挥作用的机制之一。
Objective: To investigate the effect of ERβ gene overexpression on the penile vascular endothelium in ERβKO mice and its related molecular mechanisms. Methods: Twelve ERβKO male mice were randomly divided into two groups: ERβKO + TNFα + pAdxsi-ERβ group and ERβKO + TNFα + empty virus group. ERβKO + TNFα + pAdxsi-ERβ group were transfected with ERβ gene recombinant adenovirus for 14 days, ERβKO + TNFα + empty virus group transfected with empty virus as control, while both groups were given TNFα6μg / (kg · d) intraperitoneal injection, continuous 14 d. The erectile function of mice was observed by APO method. The changes of endothelial markers CD34 and vWF were detected by immunohistochemistry. The expressions of eNOS-NO pathway related molecules were detected by RT-PCR, Western blotting and immunohistochemistry. Results: The changes of ERβKO + TNFα + pAdxsi-ERβ group were as follows: ①the number of erection increased (2.17 ± 0.41 vs 0.50 ± 0.55, P <0.05), and the latent period of erection was shortened compared with ERβKO + TNFα + (24.0 ± 1.27) min vs (28.83 ± 1.33) min, respectively, P <0.05]; ② The expression of endothelial markers CD34 and vWF were more abundant P <0.05]. ③ The expression of eNOS and Cam decreased and the expression of caveolin-1 was up-regulated (all P <0.05). The results of RT-PCR and Western blot were consistent. CONCLUSION: The ERβ gene has a protective effect on the penile vascular endothelium, and eNOS-NO pathway is one of the mechanisms.