益肾活血汤对糖尿病性ED大鼠阴茎海绵体平滑肌细胞calponin1和OPN表达的影响

The Effects of YiShenHuoXueTang on the expression of calponin 1 and OPN in corpus cavernosum smooth muscle cells of diabetic ED rats

目的 研究益肾活血汤对糖尿病性ED大鼠阴茎海绵体平滑肌细胞中碱性调宁蛋白（calponin1）和骨桥蛋白（osteopontin，OPN）表达的影响，探讨其治疗糖尿病性ED的可能机制。方法取雄性SD大鼠60只，从中随机取7只为正常对照组，余53只以链脲佐菌素（STZ）诱导建立糖尿病（DM）模型，阿朴吗啡阴茎勃起试验筛选糖尿病性ED模型后，随机选28只分为模型组、益肾活血汤组、达美康组、达美康＋益肾活血汤（联合）组，每组7只，给药12周后，应用光镜及透射电镜观察各组大鼠阴茎海绵体的形态结构。用免疫组化染色、原位杂交技术和彩色图文分析系统检测各组大鼠阴茎海绵体平滑肌细胞calponin1和0PN的表达，实时荧光定量RT-PCR分析各组大鼠阴茎海绵体平滑肌细胞calponin1mRNA和0PNmRNA的相对表达量。结果与正常对照组比较，模型组大鼠阴茎海绵体平滑肌细胞calponinl及其mRNA表达显著减少，差异均具统计学意义（P〈0．01），OPN及其mRNA表达水平显著上调，差异均具统计学意义（P〈0．01）；与模型组比较，益肾活血汤组、达美康组、联合组大鼠阴茎海绵体平滑肌细胞calponin1及其mRNA表达显著上调，OPN及其mRNA表达水平显著下降，差异均具统计学意义（P〈0．05），但与达美康组、益肾活血汤组比较，联合组calponin1及其mRNA表达水平上调较高，差异均具统计学意义（P〈0．05），同时OPN及其mRNA表达下降较多，差异均具统计学意义（P〈0．05）。联合组与正常对照组之间的差异则无统计学意义（P〉0．05）。结论益肾活血汤可以显著提高糖尿病性ED大鼠阴茎海绵体平滑肌细胞calponin1及calponinlmRNA表达水平，使平滑肌细胞表型从合成型向收缩型转化。

Objective To investigate the effect of YiShenHuoXueTang on the expression of calponin 1 and osteopontin （OPN） in corpus cavernosum smooth muscle cells of diabetic ED rats, and explore its possible mechanism in treatment of diabetic ED. Methods Among 60 male SD rats, 7 of them were set as the normal control, and the other 53 were induced to be diabetes mellitus（DM） model by streptozotocin（STZ）, and then rat models with diabetic erectile dysfunction were screened by apomorphine penis erectile test. 28 rats with diabetic erectile dysfunction were randomly recruited and divided them into the model group, YiShenHuoXueTang group, Diamicron group and Diamicron plus YiShenHuoXueTang group, there were 7 rats in each group. After 12 weeks of drug delivery, the content of the elastic fibers in the albuginea penis was observed using Light and Electron Microscopy, the expressions of calponin 1 and OPN in each group were measured by immunohistochemical staining,in situ hybridization and color image analysis. The relative expression of calponin 1 and OPN were detected by Real-time RT-PCR）. Results Compared with that of the normal control group, the expression of calponin 1 and calponin 1 mRNA in model group decreased significantly （P〈0.01, P〈0.01）, while that of OPN and OPN mRNA increased significantly （P〈0.01, P〈0.01）. Compared with that of the model group, the expression of calponin 1 and calponin 1 mRNA in YiShenHuoXueTang group, Diamicron group and Diamicron plus YiShenHuoXueTang group increased significantly, while that of OPN and OPN mRNA decreased significantly （P〈0.05）, but compared with that of Diamicron group, YiShenHuoXueTang group, the expression of calponin 1 and calponin 1 mRNA in Diamicron plus YiShenHuoXueTang group increased obviously, and there was a significant difference （P〈0.05）, simultaneously the expression of OPN and OPN mRNA reduced obviously, and there was a significant difference （P〈0.05）. There was no statistical significance between Diamicron plus YiShenHuoXueTang group and normal control group（P〉0.05）. Conclusion YiShenHuoXueTang could significantly increase the expression of calponin 1 and calponin 1 mRNA in smooth muscle cells of in corpus cavernosum tissue and transform them from contractile phenotype into synthetic phenotype.