小檗碱通过P38MAPK/ERK通路抑制阴茎海绵体细胞凋亡以增强糖尿病大鼠勃起功能

Berberine inhibited penile sponge cell apoptosis and enhanced erectile function in diabetic rats through p38MAPK/ERK pathway

目的 探讨小檗碱对糖尿病勃起功能障碍(DMED)大鼠影响及作用机制。方法 将60只SD大鼠随机分为正常组、DMEM组和小檗碱组,每组20只。除正常组外,其余各组以腹腔注射链脲佐菌素(STZ)诱导糖尿病大鼠模型,造模成功后,采用阿朴吗啡诱导阴茎勃起实验(APO)以筛选DMED大鼠模型。成模后,小檗碱组每日予以200 mg/kg的小檗碱进行灌胃,正常组和DMED组分别灌以等量生理盐水。给药4周后测定大鼠阴茎海绵体内压(ICP)和平均动脉压(MAP)以评估各组大鼠勃起功能;HE染色观察大鼠阴茎海绵体组织的病理变化;Masson染色检测阴茎海绵体组织的纤维化水平;Tunel染色检测阴茎海绵体组织细胞凋亡,Western blot检测海绵体组织中Bcl-2、Bax、Caspase-3以及p38MAPK/ERK通路相关蛋白p38MAPK(p38MAPK/p-p38MAPK)和ERK1/2(p-ERK1/2/ERK1/2)磷酸化水平。结果 与正常组比较,DMED组和小檗碱组最大ICP、MAP值和Bcl-2蛋白表达均显著降低(P<0.05),阴茎海绵体组织细胞凋亡指数、Bax、Caspase-3、p38MAPK/p-p38MAPK和p-ERK1/2/ERK1/2表达水平均显著升高(P<0.05);与DMED组相比,小檗碱组最大ICP、MAP值和Bcl-2蛋白表达水平均显著升高(P<0.05);阴茎海绵体组织细胞凋亡指数、Bax、Caspase-3、38MAPK/p-p38MAP和p-ERK1/2/ERK1/2表达均显著下降(P<0.05)。结论 小檗碱能改善糖尿病大鼠阴茎海绵体纤维化并增强勃起功能,其机制可能是通过p38MAPK/ERK通路抑制阴茎海绵体细胞凋亡实现的。

Objective To investigate the effect and mechanism of berberine on erectile function in diabetic erectile dysfunction(DMED)rats. Methods Sixty SD rats were randomly divided into normal group,DMEM group and berberine group,with 20 rats in each group. In addition to the normal group,the rest of the group with intraperitoneal injection of chain urea cephalosporins(STZ)induced diabetic rats model,after the success of the building,with apomorphine induced penile erection test(APO)in screening of DMED model in rats. After modeling,berberine group was given 200 mg/kg berberine daily intragastric administration,normal group and DMED group were given the same amount of normal saline,respectively. ICP and MAP were measured 4 weeks after administration to evaluate erectile function. HE staining was used to observe the pathological changes of the cavernosum of the penis. Masson staining was used to detect the fibrosis level of penile cavernous tissue. Tunel staining was used to detect cell apoptosis in cavernosa,Western blot was used to detect the expression of Bcl-2,Bax,Caspase-3 and phosphorylation level of p38MAPK/ERK pathway-related proteins p38MAPK(p38MAPK/p-P38MAPK)and ERK1/2(p-ERK1/2/ERK1/2)in spongy tissues. Results Compared with normal group,the maximum ICP、MAP value and Bcl-2 protein expression level in DMED and berberine groups were significantly decreased(P<0.05). Apoptosis index,Bax,caspase-3 protein expression level,p38MAPK/P-P38MAP ratio and p-ERK1/2/ERK1/2 ratio were significantly increased(P<0.05),the maximum ICP/MAP value and bcl-2 protein were significantly increased in berberine group compared with DMED group(P<0.05). Apoptosis index,Bax,caspase-3 protein expression level,p38MAPK/p-p38MAPK ratio and p-ERK1/2/ERK1/2 ratio were significantly decreased(P<0.05). Conclusion Berberine can improve diabetes rat corpus cavernosum fibrosis and enhancing erectile function and its mechanism may be through p38MAPK/ERK pathway inhibits the penis sponge body cell apoptosis.