填精通络方对糖尿病勃起功能障碍大鼠阴茎海绵体舒缩功能的影响

Effects of Tianjing Tongluo Recipe on corpus cavernosum diastolic function in rats with diabetes-induced erection dysfunction

目的:探讨填精通络方对糖尿病勃起功能障碍(DIED)大鼠阴茎海绵体舒缩功能的影响。方法:选取90只SD雄性大鼠,其中75只腹腔注射链脲佐菌素(STZ,50mg/kg)制备糖尿病模型大鼠,阿朴吗啡阴茎勃起实验筛选勃起功能障碍大鼠,将造模成功大鼠随机分为模型组,对照组(他达拉非,0.2mg/kg),中药高、中、低剂量组(填精通络方,5.40、2.70、1.35g/kg),每组10只;另选取10只未造模大鼠为正常组;各组均连续给药4周。末次给药后12h处死大鼠、取材,苏木素-伊红(HE)染色观察大鼠阴茎海绵体组织病理学改变,ELISA法检测大鼠血清中睾酮(T)、黄体生成素、促卵泡素水平,实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测大鼠阴茎海绵体组织内皮型一氧化氮合酶(eNOS)、苏氨酸激酶(Akt)、RhoA、Rho、PDE5 mRNA和蛋白表达水平,全自动生化分析仪检测各组大鼠血清Ca^(2+)水平的变化。结果:与模型组比较,各给药组大鼠首次爬背时间显著缩短(P<0.01),舔嗅次数、骑跨次数、插入次数显著增加(P<0.01),血清中T水平显著升高(P<0.01),Ca^(2+)水平显著降低(P<0.01),大鼠阴茎海绵体血管破坏,红细胞数目减少,结缔组织增生有明显改善,大鼠阴茎海绵体组织中eNOS、Akt mRNA和蛋白表达水平显著升高(P<0.01),RhoA、ROCK1、ROCK2、PDE5 mRNA和蛋白表达水平显著降低(P<0.01)。结论:填精通络中药可通过调控PI3K/Akt和RhoA/Rho信号通路,纠正大鼠阴茎海绵体收缩功能异常,改善DIED大鼠的勃起功能。

Objective: To investigate the effects of Tianjing Tongluo Recipe on the diastolic function of corpus cavernosum in diabetes-induced erection dysfunction rats. Methods: A total of 90 SD male rats were selected, and 75 rats were prepared for diabetic model by intraperitoneal injection of streptozotocin(STZ, 50 mg/kg), and erectile dysfunction rats were screened by apomorphine penis erectile test. The successful modeling rats were randomly divided into the model group, the control group(Tadalafil,0.2 mg/kg), and the high-, medium-and low-dose groups(Tianjing Tongluo Recipe, 5.40, 2.70 and 1.35 g/kg) of traditional Chinese medicine groups, with 10 rats in each group. Another 10 rats were selected into the normal group. Each group was administrated for 4 weeks. Rats were sacrificed at 12 h after the last administration. The histopathological changes of rats corpus cavernosum were observed by hematoxylin-eosin(HE) staining. The levels of testosterone(T), luteinizing hormone and follicle stimulating hormone in serum of rats were detected by ELISA. The mRNA and protein expression levels of eNOS, Akt, RhoA, Rho and PDE5 in rats corpus cavernosum were detected by real-time quantitative polymerase chain reaction(Real-time PCR) and Western blot. The change of serum Ca^(2+) level in each group was detected by automatic biochemical analyzer. Results: Compared with the model group, the first time of climbing back was significantly shortened(P<0.01), the times of licking and sniffing, the times of riding and inserting were significantly increased in treatment groups(P<0.01), the level of T in serum was significantly increased(P<0.01), the level of Ca^(2+)was significantly decreased(P<0.01), the blood vessels of corpus cavernosum were damaged(P<0.01), the number of red blood cells was decreased, the proliferation of connective tissue was significantly improved, the mRNA and protein expressions of eNOS and Akt in corpus cavernosum were significantly increased(P<0.01), and the mRNA and protein expressions of RhoA, ROCK1, ROCK2 and PDE5 were significantly decreased(P<0.01). Conclusion: Tianjing Tongluo Recipe can improve erectile function of DIED rats by regulating PI3K/Akt and RhoA/Rho signaling pathways.