基于Rho/ROCK信号通路探讨抵当汤防治糖尿病下肢动脉病变介入术后再狭窄的作用及其机制

Interventional therapy is currently a new method for the treatment of the diabetic lower extremity arterial disease (DLEAD).But restenosis after interventional treatment is the main factors influencing the curative effect. Restenosis rate every 1 years is 20-60%.The key link of restenosis after interventional therapy is postoperative vascular smooth muscle cell proliferation.Our preliminary results indicated that the Didang decoction was in accordance with postoperative DLEAD “stasis context” TCM pathogenesis characteristics.Treatment of DLEAD after intervention in patients with the Didang decoction showed clinical advantage.Whether or not the mechanism is associated with the effect on the proliferation of vascular smooth muscle cells.Study was designed to investigate the effect of Didang decoction on smooth muscle cell proliferation after vascular injury and its mechanism.Contents include:1.During the proliferation of vascular smooth muscle cells induced by H2O2,how will do change Rho/ROCK signal transduction pathway and what is its effect on the proliferation of vascular smooth muscle cells;what is effect of Didang decoction on the proliferation of vascular smooth muscle cells mediated by Rho/ROCK signal transduction pathway. 2.What is effect of Didang decoction on Rho/ROCK and vascular images of diabetic rabbits injuried by balloon.What is the molecular biological mechanism of Didang decoction influencing the proliferation of vascular smooth muscle cells after injury from the changes of Rho/ROCK signal transduction.It would provide new ideas for the prevention of restenosis after interventional therapy by classical formulae.

介入治疗是目前糖尿病下肢动脉病变（DLEAD)治疗的新方法。但介入治疗后再狭窄是影响疗效的主要因素，1年再狭窄率20-60%，5年通畅率25%。再狭窄的关键环节是术后血管平滑肌细胞过度增殖。本人前期研究结果表明抵当汤符合DLEAD术后“瘀阻脉络”中医病机特征，治疗DLEAD介入术后患者具有临床优势，其机理是否与影响血管平滑肌细胞过度增殖相关；本研究旨在探讨抵当汤对血管损伤后平滑肌细胞增殖的影响及机制。内容包括：1、H2O2诱导的血管平滑肌细胞增殖过程中，Rho/ROCK信号传导通路的变化及对血管平滑肌细胞增殖的影响，抵当汤对Rho/ROCK信号传导通路介导的血管平滑肌细胞增殖的影响。2、抵当汤对糖尿病实验兔球囊损伤血管后Rho/ROCK及血管图像的影响。从Rho/ROCK信号传导的变化探讨抵当汤影响损伤后血管平滑肌细胞增殖的分子生物学机制，为经方防治介入术后再狭窄提供新思路。

介入治疗是目前糖尿病下肢动脉病变（DLEAD)治疗的新方法。但介入治疗后再狭窄是影响疗效的主要因素，其原因在于术后血管平滑肌细胞过度增殖。本人前期研究结果表明抵当汤治疗DLEAD介入术后患者具有临床优势，本研究旨在探讨抵当汤对血管损伤后平滑肌细胞增殖的影响及机制。内容包括：1、体外实验：大鼠胸主动脉平滑肌细胞体外模型的建；抵当汤含药血清的制备；实验分组：正常对照组、模型组 、抵当汤高剂量组、抵当汤中剂量组、抵当汤低剂量组、阳性对照组；结果表明：抵当汤含药血清对A7R5细胞具有抑制作用，且高剂量抵当汤含药血清抑制作用较为明显抵当汤含药血清通过G2/M期聚集来发挥抑制细胞增殖的作用；抵当汤含药血清组也通过抑制RHO/ROCK通路发挥了作用，但是与剂量不成可比关系。2.动物实验：方法：建立糖尿病实验兔腹主动脉球囊损伤模型，分为非手术组、手术模型组、抵当汤低剂量组、抵当汤高剂量组、Y-27632组。手术前两周开始根据分组分别胃管灌饲抵当汤、Y-27632或生理盐水，球囊损伤手术后继续灌饲2周。术后通过血管影像学、病理学方法观察抵当汤对糖尿病实验兔腹主动脉球囊损伤后血管内径、内膜、中膜、血流速度等的影响。结果：抵当汤以16.2g/kg.d的剂量干预糖尿病实验兔腹主动脉球囊损伤后的血管重构起到了最大的治疗效果。其疗效在于腹主动脉球囊损伤段组织病理学的逆转、增加的动脉内径和血流速度、减轻的内膜增殖等因素。结论：这些结果证实了我们提出的假说，即抵当汤干预糖尿病实验兔腹主动脉球囊损伤后的腹主动脉壁的血管重构。下调了腹主动脉球囊损伤段RHOA/ROCK 和 PCNA 的表达，上调了损伤段P27的表达。这些结果证实了我们提出的假说，即抵当汤干预糖尿病实验兔腹主动脉球囊损伤，其机理与对RHOA/ROCK 和 P27/PCNA介导的腹主动脉球囊损伤后的血管重构相关。