降钙素基因相关肽对高血压性勃起功能障碍大鼠阴茎海绵体平滑肌细胞增殖及表型转化的影响

Effect of calcitonin gene-related peptide on proliferation and phenotype transformation of corpus cavernosum smooth muscle cells in hypertensive rats with erectile dysfunction

目的探讨降钙素基因相关肽(CGRP)对高血压性勃起功能障碍(ED)大鼠阴茎海绵体平滑肌细胞(CCSM)增殖及表型转化的影响。方法选取20只自发性高血压Wistar大鼠(SHR)及10只同系正常血压健康大鼠(WKY),测量大鼠尾部动脉收缩压,通过颈部皮下注射阿朴吗啡后观察大鼠阴茎勃起情况,SHR大鼠中无阴茎勃起的归为高血压勃起功能障碍(HBP-ED)组,出现阴茎勃起的归为高血压(HBP)组,WKY大鼠归为对照组。取各组大鼠阴茎海绵体平滑肌组织行细胞原代培养。不同浓度(1、10和100nmol/L)CGRP作用于培养的细胞后,采用MTT法检测细胞增殖情况,采用实时荧光定量PCR法检测各组细胞碱性调宁蛋白(Cnn1)和骨桥蛋白(OPN)mRNA表达水平。结果不同浓度CGRP均可抑制高血压性ED大鼠CCSM的增殖,且抑制程度随CGRP浓度增加而加剧。当CGRP作用的终浓度为100nmol/L时,HBP组和HBP-ED组Cnnl m RNA表达水平较组内0nmol/L浓度均上调(均P<0.05),OPN mRNA表达水平较组内0nmol/L浓度均下调(均P<0.05)。结论 CGRP可抑制高血压性ED大鼠CCSM的增殖,可使高血压性ED大鼠CCSM表型发生逆转化,即由合成型向收缩型转化。

Objective To investigate the effect of calcitonin gene-related peptide（CGRP） on the proliferation and phenotype transformation of corpus cavernosum smooth muscle cells（CCSM） of hypertensive rats with erectile dysfunction （ED）. Methods Male spontaneously hypertensive rats（SHR, n=20） and normotensive Wistar Kyoto rats（WKY, n=10） were used in this experiment.After measuring the systolic blood pressure of the caudal artery and examining the erectile function with subcutaneous injection of apomorphine（APO）, the rats were divided into three groups： namely control group, hypertension （HBP） group and hypertension with erectile dysfunction（HBP-ED） group. The CCSM from three groups were primarily cultured. Then the cells from three groups were exposed to 1,10 and 100nmol/L of CGRP, the proliferation of cells was measured with MTT method, and the relative expressions of calponin1（Cnn1） and osteopontin（OPN） mRNA were determined by real-time fluorescence quantitative RT-PCR. Results Different concentrations of CGRP all had an effect of inhibition on the CCSM of hyperten-sive rats with ED, and the degree of inhibition increased with the concentration of CGRP increasing. Exposure to 100nmol/L of CGRP significantly upregulated the expression of Cnnl mRNA and downregulated the expression of OPN mRNA in HBP and HBP-ED groups（all P〈0.05）. Conclusion CGRP has an inhibition effect on the CCSM in hypertensive rats with ED, and it can transform the phenotype of CCSM of hypertensive rats with ED from synthetic type to contractile type.