基于PKC通路探讨“活血通络起痿汤”改善糖尿病勃起功能障碍大鼠内皮细胞功能和血小板异常活化的机制研究

Diabetes-induced erectile dysfunction(DIED) is a common complication of ED, penile vascular endothelial injury is initiating links and pathophysiological basis of DIED. Excessive activation of PKC pathway is a key factor in diabetic vascular disease. Early application of "Huoxue Tongluo Qiwei soup" to treatment DIED achieved a significant effect which is better than “nourishing kidney and tonic yang medcine”, but the mechanism is not clear. Therefore, to explore whether it by regulating PKC pathway to improve endothelial function and then play a therapeutic role is a key problem to be solved. With this study, we use streptozotocin-induced DIED rat as models, with PKC pathway as the starting point, "Huoxue Tongluo Qiwei soup" and "PKCβ inhibitor" to intervene the models. Then measured the rats erectile function, vascular endothelial injury ,platelet activation markers and PKCβ and upstream and downstream DAG, NF-kB expression levels to explore a possible molecular mechanism for the treatment and provide a new treatment ideas and experimental evidence for application of Chinese medicine to treat DIED.

糖尿病勃起功能障碍（DIED）是男性勃起功能障碍常见类型之一，阴茎血管内皮损伤是DIED发生的始动环节和病理生理基础。PKC通路的过度激活是糖尿病血管病变的关键因素。前期应用活血化瘀方剂“活血通络起痿汤”治疗DIED效果优于传统补肾壮阳中药，但作用机制尚不明确。因此，探索本方是否通过调控PKC通路改善DIED血管内皮功能进而发挥治疗作用是研究的关键问题。本研究借助链脲佐菌素诱导的DIED大鼠模型，以PKC通路为研究的切入点，用“活血通络起痿汤”和“PKCβ”抑制剂分别进行干预，测定各组大鼠勃起功能、血管内皮损伤程度、血小板活化标志物、PKCβ及上下游位点DAG、NF-kB的表达的水平，探索本方治疗DIED可能的分子机制，为临床应用活血化瘀药物防治DIED提供新的治疗思路和实验依据。

糖尿病（DM）是一种以高血糖症为特征的慢性代谢性疾病，全球约4.51亿人患有糖尿病或有早期糖尿病风险。然而，大约50％的糖尿病男性在诊断后的10年内患有勃起功能障碍（ED）。课题组前期应用活血化瘀方剂“活血通络起痿汤”治疗DIED时发现，其治疗效果优于传统补肾壮阳中药，但作用机制尚不明确。本课题以药物干预DIED模型大鼠的实验研究为主要内容，来探讨“活血通络起痿汤”治疗DIED的潜在作用机制。实验结果发现，经药物干预后，干预组的DAG、PKCβ、NF-κB、ICAM-1的蛋白及mRNA水平均较M组有明显的下调（P＜0.05），干预组大鼠的阴茎海绵体血窦的分布较M组相对规则，内皮细胞密度增加，胶原纤维数量相对减少；超微结构上，干预组大鼠的血管内皮细胞较M组细胞核相对完整，线粒体、高尔基体、内质网等细胞器明显增多。说明活血通络起痿汤可对PKCβ信号通路起到一定的抑制作用，PKCβ信号通路的激活介导了内皮损伤、血小板异常活化等病理过程。本课题从分子层面探讨了DIED可能的发生机制，并进一步地阐明了活血通络起痿汤改善血管内皮损伤、抗血小板活化的分子机制，为该方今后的研究提供了丰富的实验数据支撑，并为临床应用活血化瘀药物防治DIED提供了新的治疗思路和实验依据。