P38丝裂素活化蛋白激酶信号通路阻断对糖尿病鼠早期血视网膜屏障和视网膜节细胞的保护作用  

Protective effect of blocking the signal path of p38 mitogen-activated protein kinase on blood-retinal barrier and retinal ganglion cells in early diabetic rats

作　　者：李永浩[1] 吕林[1] 陈凌燕[1] 黄新华[1] 张静琳[1] 李石毅[1] 王祥硅[1] 

机构地区：[1]中山大学中山眼科中心,中山大学眼科学国家重点实验室,广州510060

出　　处：《中华眼底病杂志》2010年第2期139-142,共4页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金（30672277） 广东省科技计划项目（2008B030301317,2008B030301095）

摘　　要：目的 组织化学法检测视网膜上caspase-3和血管内皮生长因子（VEGF）的荧光表达.糖尿病鼠建模后2周,对实验组行玻璃体腔注射p38 MAPK抑制剂SB203580,注射后6周检测caspase-3和VEGF的荧光表达,caspase-3免疫荧光染色法计数RGC凋亡数量.采用SPSS 13.0应用软件进行统计分析.结果 正常对照组大鼠中,IgG染色局限于血管腔内,几乎没有渗漏的迹象.糖尿病鼠建模后8周,IgG渗漏明显增加.SB203580璃体腔注射6周后的糖尿病鼠IgG渗漏减少明显.荧光免疫组织化学及相对定量分析结果显示,SB203580玻璃体腔注射6周后的糖尿病鼠视网膜上VEGF荧光表达呈下降趋势,VEGF相对荧光量从糖尿病鼠8周时较正常对照组高2.9倍减少到仅高于正常对照组1.8倍,两者比较差异有统计学意义（t=5.203,P〈0.01）.caspase-3免疫荧光染色法RGC凋亡计数结果显示,SB203580玻璃体腔注射6周后,caspase-3-阳性节细胞凋亡数与未注射SB203580相比明显减少,两者差异有统计学意义（t=5.731,P〈0.01）.结论 p38 MAPK抑制剂SB203580能够减轻糖尿病早期血视网膜屏障的破坏和视网膜节细胞的调亡,提示p38 MAPK信号通路在糖尿病早期对DR的发展起着一定的作用.Objective To investigate the protective effect of blocking the signal path of p38 mitogen-activated protein kinase on blood-retinal barrier （BRB） and retinal ganglion cells （RGC） in early diabetic rats. Methods A total of 60 Wistar rats were divided into the control and diabetes group, with 30 rats in each group. Diabetes was induced in rats in diabetes group by peritoneal injection of streptozotocin （STZ） the plasma glucose level of 〉16.7 mmol/L indicated that the diabetes model was set up successfully. The rats in the control group underwent peritoneal injection of equivalent sodium citrate solution. IgG leakage method was used to measure the damage of BRB function and vascular leakage. The expression and localization of caspase-3 and vascular endothelial growth factor （VEGF） in retina of diabetic rats were examined by immunohistochemistry analyses. Two weeks after the establishment of the diabtes model, the rats in diabtes group underwent intravitreal injection with SB203580, a p38 inhibitor six weeks after the injection, the expression of caspase-3 and VEGF was detected, and the number of apoptosis RGC was counted via immunofluorescence technique. Results In the contral group, IgG staining located in the blood vessels with little leakage while the IgG leakage was much more obvious in the diabetes group eight weeks after the establishment of the model. Six weeks after intravitreal injection with SB203580, the leakage decreased in diabtes rats. The results of semi-quantitative analysis and fluorescence immunohistochemistry showed that compared with the results in diabetes rats 8 weeks after intravitreal injection （2.9 times much more than that in the control group）, the fluorescence expression of VEGF decreased in diabetes rats six weeks after intravitreal injection （1.8 times much more than that in the control group）. The apoptisis RGC number in rats 6 weeks after intravitreal injection of SB203580 was much less than that in rats without intravitreal injection （t=5. 731, P〈0.

关 键 词：糖尿病视网膜病变/病理生理学 p38丝裂原活化蛋白激酶类/生理学 血视网膜屏障/生理学 视网膜神经节细胞/生理学 糖尿病 实验性 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]