染料木黄酮防治大鼠移植动脉硬化的实验研究

Experimental study of genistein in preventing and treating transplant arteriosclerosis in arotic transplantation of rat

目的探讨染料木黄酮(Genistein,GEN)对移植动脉硬化的保护作用及其机制。方法建立Brown-Norway至Lewis大鼠腹主动脉移植模型,将受体鼠随机分为四组:1组)、(n=8):高剂量染料木黄酮组(20mg/kg·d);2组(n=8):高低剂量结合的染料木黄酮组[20mg/(kg·d)14d+2mg/(kg·d)46d]腹腔注射60d染料木黄酮(GEN);3组(n=5):溶剂对照组,腹腔注射溶剂二甲基亚砜(DMSO)60d;4组(n=5):异系对照组,不给予任何处理。移植后60d取移植血管作病理组织学检查并利用算机图像分析系统测量内膜厚度;免疫组织化学检测CD4+、CD8+T淋巴细胞的浸润;ELISA法检测血清中血管内皮生长因子(VEGF)的变化。结果病理组织学检查两实验组分别与两对照组比较,内膜增厚不明显(P<0.001)且淋巴细胞和巨噬细胞浸润减少,免疫组织学检测CD4+、CD8+T淋巴细胞浸润减少(P<0.01);两对照组血清中VEGF表达显著高于实验1组即高剂量组(P<0.01),但与实验2组即高低剂量结合组无明显差别。结论染料木黄酮对移植动脉硬化有保护作用,其机制可能与减少了淋巴细胞的浸润,影响生长因子的表达有关,但其主要作用机制有待进一步研究。

[Objective ] To study the protective effects of genistein（GEN） in preventing and treating transplant arteriosclerosis and potential mechanisms. [Methods] Arotic transplantation was performed with male Brown-Norway rats as donors and male Lewis rats as recipients. The recipients in the experiment was randomly divided into four groups： （1） high--dose group （n=8）, the recipients were administered with GEN at a dose of 20 mg/kg body weight for 60 days continuously; （2） high-low dose group （n=8）, at a dose of 20 mg/kg for 14 days and at dose of 2 mg/kg for 46 days continuously; （3） solvent control （n=5）, 0.5 mL DMSO for 60 days; （4） control （n=5）, no treatment. Histological changes were monitored 60 days after transplantation, and the average depth of intima was calculated with computer imaging analysis systes. Immunohistochemistry was used to detect infiltration of CD4^＋, CD8^＋ T lymphocyte in the transplanted vessels, and ELLSA to detect the change of VEGF in different group. [Results] Compared with two control groups, there were less lymphocytes and macrophages infiltration, the average depth of intima in two experimental groups markedly lowed （seperatedly P 〈0.001） and infiltration of CD4^＋, CD8^＋ T lymphocyte in the transplanted vessels was less （seperatedly P 〈0.01）. Expression of VEGF in high dose group was lower than that in two control （8.6731±1.0466 Vs 21.8340±0.7558 and 23.2068±1.2028 P 〈0.01）, but there was no statistic difference compared with high-low dose group. [Conclusion] Genistein can prevent the development of graft arteriosclerosis and the lowed infiltration of lymphocytes and the influenced expression of growth factors may be involved in the mechanisms.