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目的探讨糖尿病大鼠尿道α1肾上腺素能受体与神经生长因子(NGF)表达的改变及其对尿道功能的影响。方法 20只雌性Wistar大鼠随机分为正常对照组(10只)和糖尿病组(10只)。采用尿流动力学方法检测大鼠膀胱与尿道压力;RT-q PCR与Western blotting测定两组大鼠尿道中α1肾上腺素能受体的表达;RT-q PCR与ELISA法测定两组大鼠尿道中NGF的表达。Western blotting测定两组大鼠尿道中NGF前体(pro NGF)、P75神经营养因子受体(P75NTR)及Sortilin的表达水平。结果与对照组比较,糖尿病组大鼠尿道舒张时的最低尿道灌注压增高;静脉注射α1肾上腺素能受体拮抗剂(盐酸坦索罗辛)可降低糖尿病大鼠的最低尿道灌注压以及尿道灌注压的基线水平。RT-q PCR与Western blotting结果显示,糖尿病组大鼠尿道α1a和α1d受体表达水平升高(P<0.05)。RT-q PCR与ELISA结果显示,糖尿病组大鼠尿道NGF的表达明显减少。Western blotting结果显示,糖尿病组大鼠pro NGF的表达升高(P<0.05)、大鼠尿道中P75NTR表达减少(P<0.05),两组比较Sortilin的表达差异无统计学意义(P>0.05)。结论糖尿病大鼠尿道中α1肾上腺能受体表达水平的升高以及NGF/pro NGF通路的改变可能是导致糖尿病尿道功能紊乱的机制。
Objective To investigate the changes of urethral α1-adrenergic receptor and nerve growth factor (NGF) expression in diabetic rats and their effects on urethral function. Methods Twenty female Wistar rats were randomly divided into normal control group (n = 10) and diabetic group (n = 10). Urinary flow dynamics was used to detect bladder and urethral pressure in rats. RT-q PCR and Western blotting were used to detect the expression of α1-adrenergic receptor in the urethra in both groups. RT-q PCR and ELISA were used to determine the urinary tract NGF expression. Western blotting was used to detect the expression of pro-NGF, P75NTR and Sortilin in the urethra of the two groups. Results Compared with the control group, the minimum urethral perfusion pressure was increased in the urethral diastole of diabetic rats. Intravenous α1 adrenergic receptor antagonist (tamsulosin hydrochloride) could reduce the lowest urinary perfusion pressure and urethral perfusion in diabetic rats Pressure baseline level. RT-q PCR and Western blotting results showed that urethral α1a and α1d receptor expression in diabetic rats increased (P <0.05). The results of RT-q PCR and ELISA showed that the expression of NGF in urethra of diabetic rats decreased significantly. The results of Western blotting showed that the expression of pro NGF in diabetic rats increased (P <0.05) and the expression of P75NTR in urethra decreased (P <0.05). There was no significant difference in the expression of Sortilin between the two groups (P> 0.05) . Conclusion The increased expression of α1 adrenergic receptor in the urethra and the alteration of NGF / pro NGF pathway may be the mechanism of diabetic urethral dysfunction.