塞来昔布下调JAML抑制糖尿病视网膜病变大鼠VEGF的表达及其机制  

Mechanism of celecoxib inhibiting the expression of retinal VEGF in diabetic retinopathy rats via JAML

作　　者：段梅 曹凡 桂衍超 陈可洋[2] 陶黎明[1] 蒋正轩[1] Duan Mei;Cao Fan;Gui Yanchao;Chen Keyang;Tao Liming;Jiang Zhengxuan(Dept of Ophthalmology,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601;Dept of Inspection and Quarantine Health,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学第二附属医院眼科,合肥230601 [2]安徽医科大学卫生检验与检疫教研室,合肥230032

出　　处：《安徽医科大学学报》2023年第8期1293-1299,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82070986、82171043);安徽省学术和技术带头人科研活动经费资助项目(编号:2021H255)。

摘　　要：目的探讨塞来昔布对糖尿病视网膜病变(DR)大鼠视网膜血管内皮生长因子(VEGF)的作用及其机制。方法45只SD大鼠,随机分为正常对照(NC)组、DR组、塞来昔布干预DR(DR+C)组。腹腔注射1%链尿佐菌素(STZ)建立糖尿病模型。1个月后,DR+C组给予塞来昔布(50 mg/kg)灌胃(1次/d)。2个月后检测血清总胆固醇(TC)、胰岛素(Insulin);观察视网膜组织病理特征;Western blot检测视网膜连接黏附分子样蛋白(JAML)、VEGF及其信号通路各蛋白的表达;检测视网膜白介素(IL)-10、血管细胞黏附分子-1(VCAM-1)等蛋白分布。HUVEC细胞分为正常葡萄糖组(NG)组、高糖(HG)组和高糖加药塞来昔布(HG+C)组,检测上述蛋白的表达水平。结果与DR组相比,DR+C组大鼠视网膜变薄;与NC组比较,DR+C组大鼠视网膜增厚。DR+C组视网膜JAML、磷脂酰肌醇激酶3(PI3K)、磷酸化磷脂酰肌醇激酶-3(P-PI3K)、缺氧诱导因子1-α(HIF1-α)、VEGF等蛋白低于DR组,高于NC组。DR+C组与DR组比较视网膜IL-10、VCAM-1表达下降。DR+C组和DR组TC含量高于NC组(P<0.01);DR+C组和DR组Insulin含量低于NC组(P<0.001)。与HG组相比,HUVEC细胞中HG+C组JAML、P-PI3K、HIF1-α、VEGF等表达下降(P<0.05),但高于NG组(P<0.05);三组细胞PI3K蛋白差异无统计学意义。结论塞来昔布能降低DR大鼠视网膜VEGF和炎症因子IL-10、VCAM-1的表达水平,其机制可能与JAML介导的PI3K/HIF1-α信号通路有关。Objective To investigate the effect and mechanism of celecoxib on retinal vascular endothelial growth factor(VEGF)in rats with diabetic retinopathy.Methods Forty-five SD rats were divided into normal control group(NC),diabetic retinopathy group(DR),celecoxib intervention diabetic retinopathy group(DR+C).The diabetic model was established by intraperitoneal injection of 1%STZ.After one month,celecoxib(50 mg/kg)was given intragastric administration(1/day)in the DR+C group.Two months later,serum total cholesterol(TC)and insulin were detected.The histopathological changes of the retina were observed.The expression of junctional adhesion molecule-like protein(JAML),VECF and their signal pathway proteins and the distributions of interleukin-10(IL-10),vascular cell adhesion molecules-1(VCAM-1)were detected by Western blot.HUVEC cells were divided into normal glucose group(NG),high glucose group(HG)and high glucose plus celecoxib group(HG+C)to detect the expression of the above proteins.Results Compared with DR,retina in DR+C group was thinner.The retina in the DR+C group was thicker than that in the NC group.The levels of retinal JAML,phosphatidylinositol kinase3(PI3K),phosphorylphosphatidylinositol kinase3(P-PI3K),hypoxia-inducible factorl-α(HIF1-α),and VECF in DR+C group were lower than those in DR group,while higher than those in NC group.The expression of retinal IL-10 and VCAM-1 decreased.The content of TC in DR+C and DR group was higher than those in NC group(P<0.O1),while the content of insulin in DR+C and DR group was lower than thlse in NC group(P<0.001).Compared with HG group,the expressions of JAML,PI3K,P-PI3K,HIF1-α,VEGF in HG+C group decreased,but was higher than those in NG group.There was no significant difference in PI3K among the three groups.Conclusion Celecoxib can decrease the expression of VECF,IL-10,VCAM-1 in retina of DR rats,which may be related to the PI3K/HIF1-αsignaling pathway mediated by JAML.

关 键 词：塞来昔布 JAML 糖尿病视网膜病变 磷脂酰肌醇 

分 类 号：R774.1[医药卫生—眼科]