塞来昔布对糖尿病大鼠视网膜缝隙连接蛋白Cx43的作用

Effect of celecoxib on the expression of connexin43 in retina of diabetic rats

目的:观察COX-2(cyclooxygenase,COX-2)抑制剂(塞来昔布)对糖尿病大鼠视网膜缝隙连接蛋白Cx43(connexin43,Cx43)表达的影响。方法:选取SD(Sprague-Dawley)大鼠45只,随机分为3组:对照组、糖尿病组、用药组,每组各15只,采用STZ腹腔注射造模的方式进行造模,快速血糖仪监测各组大鼠空腹血糖,正常对照组、糖尿病模型组、用药组分别用生理盐水、生理盐水、塞来昔布溶液进行灌胃,3mo后采用过量麻醉法处死大鼠,制备视网膜标本,采用免疫组化法观察Cx43蛋白,实时定量PCR技术检测大鼠视网膜Cx43mRNA的相对表达量,采用SPSS13.0统计软件进行统计分析,对各组检测结果进行比较。结果:免疫组织化学染色缝隙连接蛋白Cx43在视网膜大鼠的神经节细胞层、神经纤维层、内丛状层、色素上皮层、内皮细胞层可见表达不同程度点状、片状表达,计算机图像灰度分析发现塞来昔布对糖尿病大鼠视网膜Cx43表达有促进作用,正常对照组、糖尿病组、用药组灰度值分别是0.233±0.025、0.124±0.014、0.197±0.021,两两比较差异均有统计学意义(P<0.05);实时定量PCR检测发现塞来昔布促进糖尿病大鼠Cx43 mRNA表达量增加,正常对照组、糖尿病组、用药组相对表达量分别是0.635±0.084、0.110±0.061、0.367±0.074,两两比较差异有统计学意义(P<0.05)。结论:缝隙连接蛋白Cx43在糖尿病大鼠视网膜中表达减少,塞来昔布可以减缓糖尿病大鼠视网膜缝隙连接蛋白Cx43表达的减少。

AIM： To investigate the expression of connexin43（ Cx43） in retina of Sprague Dawley（ SD） rats with diabetes mellitus（ DM）,and the effects of celecoxib,inhibitor of cyclooxygenase 2（ COX-2）,on expression of Cx43 in retina of them. METHODS： DM was induced in SD rats by the administration of streptozotocin（ STZ）. DM rats were divided into two groups： COX-2 treated group（ CTG,n =15）,and diabetic group（ DG,n = 15）. Normal control group（ NCG） consisted of 15 rats was given. Three months＇ treatment was given after DM development.Sample of retina were made at the end of 3 mo.Expression of Cx43 in retina were investigated by immunohistochemistry. Expression of Cx43 mRNA in retina were analyzed by RT-PCR. RESULTS： Cx43 was expressed in the ganglion cell layer,the nerve fiber layer,the inner plexiform layer,and the pigment epithelium layer of retina with a different degree. Computer image analysis showed that celecoxib had elevated the expression of Cx43 in experimental diabetic rats,gray values were： 0. 233 ± 0. 025,0. 124 ±0. 014, 0. 197 ± 0. 021; multiple comparisons found statistically significant difference（ P〈0. 05）. Celecoxib had raised the expression of Cx43 mRNA in diabetic rats with real time quantitative PCR. Relative value was 0. 635 ±0. 084,0. 110±0. 061,0. 367±0. 074,multiple comparisons found statistically significant difference（ P〈0. 05）.CONCLUSION： Expression of Cx43 decreases in retina of SD rats with DM. Celecoxib can relieve the decrease of Cx43 expression in retina of them.