益气活血法通过PKG/Myocardin抑制缺氧诱导海绵体平滑肌细胞合成型转化的机制研究

Hypoxia induced corpus cavernosum smooth muscle cell phenotype transformation is the main cause of its dysfunction. It's the common pathophysiological basis in patients with impotence. Traditional Chinese medicine holds that long illness leading to qi deficiency. All kinds of pathogenic factors can cause qi and blood performing poorly in penis, or even blood stasis blocking in the penis. Qi deficiency can't regulate blood and blood stasis cause to impotence. Thus qi deficiency and blood stasis is the ultimate pathogenesis of impotence. Clinical formular Rhodiola rosea I that reinforcing qi and activating blood can effectively improve the erectile hardness of patients with impotence. Research found that it can improve the rat’s corpus cavernosum smooth muscle cell phenotype transformation induced by hypoxia, but the mechanism is unclear. This project adopts the bilateral cavernous nerve resection in rats to establish impotence model, and with the intervention of formular Rhodiola rosea I. The primal generation of rat’s corpus cavernosum smooth muscle cell will be culruted. Hypoxia model will be established with hypoxia chamber, and with the intervention of medicated serum. PKG and Myocardin will be silenced and overexpressed respectively. Detection of phenotypic regulation factors as PKG, Myocardin, SRF and phenotypic proteins as alpha SMA, SM - 22 alpha, Vimentin, Desmin, Calponin 1 in vivo and in vitro. To clarify the molecular mechanism of formular Rhodiola rosea I inhibiting the corpus cavernosum smooth muscle cell phenotype transformation induced by hypoxia through the PKG/Myocardin pathway. For providing theoretical basis for the prevention and treatment of impotence with the reinforcing qi and activating blood decoction.

缺氧诱导海绵体平滑肌细胞（SMC）表型转化是SMC功能紊乱的主要原因，是阳痿患者的共同病理生理学基础。中医认为病久气虚，各种致病因素都可导致阴茎气血运行不畅，甚或瘀血阻滞于阴茎脉络，气虚运血无力、血行瘀滞而成阳痿，因此气虚血瘀是阳痿的最终病机。临床益气活血的红景I号方能有效改善阳痿患者的勃起硬度，改善缺氧诱导的大鼠海绵体SMC表型转化，但机制不明确。本项目采用大鼠双侧海绵体神经切除建立阳痿模型，红景Ⅰ号干预；以大鼠原代海绵体SMC为靶细胞，缺氧小室建立缺氧模型，红景Ⅰ号含药血清干预，分别沉默及过表达PKG、Myocardin；体内外检测表型调控因子PKG、Myocardin、SRF和表型蛋白α-SMA、SM-22α、Desmin、Calponin 1、Vimentin，明确红景Ⅰ号通过PKG/Myocardin抗缺氧诱导海绵体SMC表型转化的分子机制，为中药益气活血防治阳痿提供理论依据。

海绵体平滑肌功能紊乱是阳痿患者的病理生理学共性，而缺氧是导致海绵体平滑肌功能紊乱的关键因素。中医认为气虚运血无力、血行瘀滞而成阳痿，气虚血瘀是阳痿的最终病机。益气活血的红景I号方能有效改善阳痿患者的勃起硬度，改善缺氧诱导的大鼠海绵体SMC表型转化，但机制不明确。本项目采用大鼠双侧海绵体神经损伤建立阳痿模型，红景Ⅰ号干预，检测大鼠勃起功能，海绵体内压，TGF-β1、Collogen I、Collogen III蛋白，表型调控因子PKG、Myocardin和表型蛋白α-SMA、Desmin、Vimentin，红景Ⅰ号通过Myocardin抗缺氧诱导海绵体SMC表型转化的分子机制。 .研究表明双侧海绵体神经（CN）损伤大鼠海绵体内压显著下降，而中、高剂量红景I号方可改善双侧CN损伤大鼠勃起功能。CN损伤大鼠海绵体组织中平滑肌与胶原纤维含量比例显著下降、胶原纤维含量增多、平滑肌含量减少。而红景I号方干预治疗后，中、高剂量组大鼠组织结构得到明显的改善。CN损伤大鼠海绵体组织中HIF-1α显著上升，红景I号方干预后HIF-1α表达明显下降。此外，CN损伤大鼠海绵体组织中TGF-β1、Collogen I、Collogen III阳性表达上升明显，红景I号方干预后可一定程度抑制TGF-β1、Collogen I 、Collogen III表达。CN损伤大鼠海绵体中收缩型蛋白α-SMA、Desmin表达明显下降，合成型蛋白Vimentin表达明显上调，即大鼠海绵体平滑肌发生表型转化。CN损伤大鼠PKG、Myocardin表达均下降明显，红景I号干预后PKG、Myocardin表达增加。红景I号方干预后通过PKG、Myocardin抑制了海绵体平滑肌细胞表型转化，为中药益气活血防治阳痿提供理论依据，可应用于临床。