生精胶囊通过调节一氧化氮诱导阴茎海绵体平滑肌舒张改善去势大鼠勃起功能

Shengjing Capsule improves the erectile function by regulating NO-induced relaxation in corpus cavernosum smooth muscle in a castrated rat model

目的探讨生精胶囊改善雄性去势大鼠勃起功能的潜在作用机制。方法2015年4—6月将3个月龄雄性SD大鼠40只,按随机数字法随机分为假手术组、去势组、生精胶囊组和睾酮组。假手术组作为正常对照,其余组均行双侧睾丸切除术建立勃起功能障碍模型。术后假手术组、去势组给予蒸馏水每天灌胃1次,生精胶囊组给予310.2mg/(kg·d)剂量的生精胶囊混悬液每天灌胃1次,睾酮组给予十一酸睾酮26.3mg/kg肌注4周1次。连续干预8周后,剖取阴茎海绵体组织,Massons三色染色法检测平滑肌与胶原纤维的比例;免疫组织化学技术检测α-肌动蛋白(α-SMA)和碱性调宁蛋白(calponin)的表达;逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹法(Western blot)检测神经型一氧化氮合酶(nNOS)、诱导型一氧化氮合酶(iNOS)、内皮型一氧化氮合酶(eNOS)的表达。结果生精胶囊组大鼠阴茎海绵体平滑肌与胶原纤维的比例为1.03±0.091,显著高于去势组0.11±0.025(P=0.000)。生精胶囊组阴茎海绵体组织α-SMA和calponin表达量分别为(16.51±0.79)%、(20.84±1.36)%,明显高于去势组(7.37±0.59)%、(1.25±1.03)%(P=0.000,P=0.000)。生精胶囊组阴茎海绵体组织nNOS、iNOS、eNOS的mRNA和蛋白表达均显著高于去势组(P值均<0.05)。结论生精胶囊可能通过保护阴茎海绵体组织含量以及增加海绵体组织一氧化氮合酶活性从而改善去势ED大鼠的勃起功能。

Objective To investigate the underlying mechanism of Shengjing Capsule in the improvement of erectile function in a male castrated rat model. Methods A total of 40 adult male Sprague-Dawley rats were randomly divided into 4 groups：sham-operated group,castration group,Shengjing Capsule group and testosterone group. Sham-operated group served as normal controls,while other groups were subjected to bilateral orchiectomy to construct models of erectile dysfunction （ED）. Rats in the sham-operated group and castration group were treated with distilled water by gavage,while rats in the Shengjing Capsule group and testosterone group were given 310.2 mg/（kg · day） Shengjing Capsule suspension by gavage and intramuscularly injected with 26.3 mg/（kg · month） testosterone undecanoate, respectively. After 8 weeks, the rats＇ penile tissues were harvested. The ratio of smooth muscle cell/collagen fibril （SMC/CF） was evaluated with Masson＇s trichrome staining. The expressions of smooth muscle alpha-actin （α-SMA） and calponin were detected with immunohistochemical staining. The expressions of neuronal nitric oxide synthase （nNOS）,inducible NOS （iNOS） and endothelial NOS （eNOS） in cavernous tissues were detected with real time reverse transcription PCR and Western blotting. Results The SMC/CF ratio was significantly higher in Shengjing Capsule group （1. 034±0. 091） than in the castration group （0.11±0. 025） （P=0. 000）. The Shengjing Capsule group had markedly higher expressions of α-SMA and calponin [（16.51 ±0.79）%, （20.84±1.36）%] than the castration group [（ 7.37 ± 0.59 ） %, （ 1.25 ± 1.03 ）%], （ P = 0.000, P = 0.000）. The gene and protein expressions of nNOS, iN OS, eN OS in cavernous tissues in Shengjing Capsule-treated rats were significantly higher than in the castrated models （all P〈0. 05）. Conclusion Shengjing Capsule improves the erectile function by protecting the smooth muscle content and enhancing NOS activity in penile tissues of castrated male rats.