糖尿病大鼠视网膜血管内皮细胞p53、bcl-2及生长因子的表达与细胞周期阻滞  

The expression of p53, bcl-2, and growth factors and cell cycle arrest of retinal vascular endothelial cells of diabetic rats

作　　者：李爱冬[1] 张晓[1] 羊惠君[2] 韦纯义[3] 

机构地区：[1]第三军医大学成都军医学院分子生物实验中心,成都610083 [2]四川大学华西医学中心解剖教研室 [3]四川大学华西医院眼科

出　　处：《中华眼底病杂志》2003年第1期29-33,共5页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的　观察 p5 3、bcl- 2、bax基因、血管内皮细胞生长因子 (vascular endothelial cell growth fac-tor,VEGF)、碱性成纤维细胞生长因子 (basic fibroblastgrowth factor,b FGF)、胰岛素样生长因子 - I(insulin-like growth factor- I,IGF- I)及其受体在 1～ 2 0周糖尿病大鼠视网膜血管内皮细胞 (vascular endothelial cell,VECs)的表达以及与细胞周期阻滞的关系。　方法　四氧嘧啶 (alloxan)诱导制作糖尿病大鼠模型 ;免疫组织化学染色 ,光、电镜观察 ;斑点印迹 (dot blotting)测定 p5 3、bcl- 2 m RNA;Western印迹 (Western blotting)测定p5 3、bcl- 2蛋白。　结果　免疫组织化学染色后光学显微镜观察显示 8～ 2 0周糖尿病大鼠视网膜节细胞层、内核层的各类血管都呈 p5 3、bcl- 2免疫阳性反应。电镜观察显示阳性反应物沉淀在 VECs。VECs同时为 VEGF、b FGF、IGF- I蛋白及其受体阳性反应。视网膜内其它细胞及同窝对照大鼠和 1～ 6周病程糖尿病大鼠视网膜所有细胞均未见上述蛋白阳性反应。 Dot blotting测定结果显示 ,2 0周糖尿病大鼠视网膜组织有 p5 3和 bcl- 2m RNA表达。Western blotting测定证实 2 0周糖尿病大鼠视网膜表达 p5 3和 bcl- 2蛋白。对照组及 1～ 2 0周糖尿病大鼠视网膜内未见Objective To observe the expression of p53, bcl 2 genes, vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin like growth factor I (IGF I), and the receptors of these factors of retinal vascular endothelial cells (VECs) of 1 to 20 week diabetic rats, and the relationship between the expressions and cell cycle arrest. Methods Retinal sections of diabetic rats induced by alloxan were immunohistochemically stained and observed by light microscopy (LM) and electron microscopy (EM). Dot blotting and Western blotting were used to determine the expression of mRNA, proteins of p53 and bcl 2. Results Under LM, immunohistochemical positive expression of p53 and bcl 2 were found on the vessels of ganglion cell layer and inner nuclear layer of retinae of 8 to 20 week diabetic rats; under EM, these substances were observed depositing in VECs. The retinal VECs also expressed VEGF, bFGF, IGF I and their receptors. There was no positive expression of other cell types in these retinae, all cell types of retinae in control group, or all cells of retinae of diabetic rats with the course of disease of 1 to 6 weeks. The result of dot blotting revealed that retinal tissue of 20 week diabetic rat expressed p53 and bcl 2 mRNA, and the result of Western blotting revealed that they also expressed p53 and bcl 2 proteins. But retinal tissues of control group did not. Positive expression of bax was not found in the retinae in control group or 1 to 20 week diabetic rats. Conclusion p53, bcl 2 may introduce cell cycle arrest of VECs of retinae in 8 to 20 week diabetic rats. High glucose might stimulate the expression of VEGF, bFGF, IGF I and their receptors, and the growth factors may keep VECs surviving by self secretion.

关 键 词：细胞周期阻滞 糖尿病视网膜病 血管内皮 免疫组织化学 视网膜血管 内皮生长因子 BCL-2基因 p53基因 

分 类 号：R774.1[医药卫生—眼科]