抑制NADPH氧化酶信号通路：葫芦巴丸改善糖尿病勃起功能障碍的分子机制

With the increase of peoples demand for higher living quality, diabetic erectile dysfunction (DMED) has become a rising concern, it is imperative to develop new drug which integrates lowering glycemia and improving DMED. Previous studies have shown that fenugreek pill and effective components can inhibit protein expression of NADPH oxidase in islet cells and testicular tissue, thus improving oxidative-stress damage of the organization. Recent studies found that the inhibition of NADPH oxidase expression can reduce the production of peroxides in the penile corpus cavernosum of diabetic rats, and then improve penile erectile function. Based on the previous research and the latest findings, we hypothesize that fenugreek pill improve DMED through inhibiting NADPH oxidase signaling pathway in penile corpus cavernosum. In this project, we will establish the model of diabetic ED to examine the levels of blood glucose and insulin sensitivity by combining with the animal and cell experiment. For mechanistic study, we will examine NADPH oxidase expression in corpora cavernosa, as well as subsequent changes in the injury of the endothelium and smooth muscle in corpora cavernosa and the apoptosis of cavernosa tissue. Different experimental techniques including immunofluorescence, real time PCR and Western blot. The project will explore that the molecular mechanism of fenugreek pill improving DMED is possibly NADPH oxidase-related oxidative stress. Additionally, the proposed research will provide new ideas to find a traditional Chinese medicine that integrates lowering glycemia and improving DMED.

随着人们对生活质量要求的提高，糖尿病勃起功能障碍（DMED）日益受到关注，寻找集降糖与改善DMED于一体的药物势在必行。前期研究显示：葫芦巴丸及有效成分可抑制胰岛细胞及睾丸组织中NADPH氧化酶蛋白表达，从而改善活性氧族对组织的氧化应激损伤。近期研究发现抑制NADPH氧化酶蛋白表达，可减少糖尿病大鼠阴茎海绵体内过氧化物的产生，使勃起功能得到改善。本项目基于前期研究和最新发现，提出“抑制NADPH氧化酶信号通路是葫芦巴丸改善DMED的分子机制”的假说。本课题采用动物实验和细胞实验相结合，在检测糖代谢和胰岛素敏感性的同时，采用免疫荧光、RT-PCR、Western Blot等方法，检测阴茎海绵体NADPH氧化酶分子表达、氧化应激引起的海绵体内皮、平滑肌损伤和组织凋亡，探讨葫芦巴丸抑制NADPH氧化酶介导的氧化应激改善DMED的可能性，为寻找具有降糖与改善DMED双重功效的中药提供了新思路。

随着人们对生活质量要求的提高，糖尿病勃起功能障碍（DMED）日益受到关注，寻找集降糖与改善DMED于一体的药物势在必行。前期研究显示：葫芦巴丸及有效成分可抑制胰岛细胞及睾丸组织中NADPH氧化酶蛋白表达，从而改善活性氧族对组织的氧化应激损伤。近期研究发现抑制NADPH氧化酶蛋白表达，可减少糖尿病大鼠阴茎海绵体内过氧化物的产生，使勃起功能得到改善。本项目基于前期研究和最新发现，提出“抑制NADPH氧化酶信号通路是葫芦巴丸改善DMED的分子机制”的假说。本课题采用动物实验，在检测糖代谢和胰岛素敏感性的同时，采用免疫荧光、RT-PCR、Western Blot等方法，检测阴茎海绵体NADPH氧化酶分子表达、氧化应激引起的海绵体内皮、平滑肌损伤和组织凋亡，探讨葫芦巴丸抑制NADPH氧化酶介导的氧化应激改善DMED的可能性，为寻找具有降糖与改善DMED双重功效的中药提供了新思路。与模型组相比，治疗组大鼠丙二醛（MDA）和过氧化物歧化酶（SOD）水平明显改善。与模型组相比，治疗组大鼠海绵体内压与颈动脉压之比上升，说明葫芦巴丸治疗后大鼠的勃起功能有所改善。与模型组相比，治疗组大鼠阴茎组织结构可见明显改善。与模型组相比，治疗组α-SMA与nNOS mRNA和蛋白表达明显提高，P47phox mRNA和蛋白表达明显降低。本实验研究结果证实葫芦巴丸能通过抑制NADPH氧化酶改善糖尿病大鼠勃起功能障碍。