塞来昔布对糖尿病大鼠视网膜缝隙连接蛋白Cx43的作用
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摘要
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.
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.
引文
1 Eisma JH,Dulle JE,Fort PE.Current knowledge on diabetic retinopathy from human donor tissues.World J Diabetes 2015;6(2):312-320
2 Wan TT,Li XF,Sun YM,et al.Recent advancesin understanding the biochemical and molecular mechanism of diabetic retinopathy.Biomed Pharmacother 2015;74(3):145-147
3 Zheng L,Howell SJ,Hatala DA,et al.Salicylate-based antiinflamatory drus inhibit the early lesion of diabetic retinopathy.Diabetes2007;56(2):337-345
4 Bobble MW,Roy S,Trodeau K,et al.Reduced connexin 43 expression and its effect on the development of vascular lesions in retinas of diabetic mice.Invest Ophthalmol Vis Sci 2010;51(7):3758-3763
5 Tien T,Barrette KF,Chronopoulos A,et al.High glucose alters Cx43expression and gapjunction intercellular communication in retinal müller cells:promotes Müller cell and peficyte apoptosis.Invest Ophthalmol Vis Sci 2014;55(7):4327-4337
6 Tien T,Muto T,Zhang J,et al.Association of reduced Connexin 43expression with retinal vascular lesions in human diabetic retinopathy.Exp Eye Res 2016;146:103-106
7 孙国玲,李筱荣,孙靖.环氧化酶-2在早期糖尿病大鼠视网膜中表达的研究.天津医科大学学报2008;14(3):335-338
8 EL-Asrar AM,Missotten L,Geboes K.Expression of cyelooxygenase-2and downstream enzymes in diabetic fibrovascular epiretina I membranes.Br J Ophthalmol 2008;92(11):1534-1539
9 Schoenberger SD,Kim SJ,Sheng J,et al.Increased prostaglandin E2(PGE2)levels in proliferative diabetic retinopathy.and correlation with VEGF and inflammatory eytokines.Invest Ophthalmol Vis SCi 2012;53(3):5906-5911
10 Tien T,Muto T,Barrette K,et al.Downregulation of Connexin 43promotes vascular cell loss and excess permeability associated with the development of vascular lesions in the diabetic retina.Mol Vis 2014;20(12):732-741
11 Li K,Yao J,Shi L,et al.Reciprocal regulation between proinflammatory cytokine-induced inducible NO synthase(iN OS)and connexin43 in bladder smooth muscle cells.J Biol Chem 2011;286(48):41552-41562
12 Tsuchida S,Arai Y,Kishida T,et al.Silencing the expression of connexin 43 decreases inflammation and joint destruction in experimental arthritis.J Orthop Res 2013;31(4):525-530
2 Wan TT,Li XF,Sun YM,et al.Recent advancesin understanding the biochemical and molecular mechanism of diabetic retinopathy.Biomed Pharmacother 2015;74(3):145-147
3 Zheng L,Howell SJ,Hatala DA,et al.Salicylate-based antiinflamatory drus inhibit the early lesion of diabetic retinopathy.Diabetes2007;56(2):337-345
4 Bobble MW,Roy S,Trodeau K,et al.Reduced connexin 43 expression and its effect on the development of vascular lesions in retinas of diabetic mice.Invest Ophthalmol Vis Sci 2010;51(7):3758-3763
5 Tien T,Barrette KF,Chronopoulos A,et al.High glucose alters Cx43expression and gapjunction intercellular communication in retinal müller cells:promotes Müller cell and peficyte apoptosis.Invest Ophthalmol Vis Sci 2014;55(7):4327-4337
6 Tien T,Muto T,Zhang J,et al.Association of reduced Connexin 43expression with retinal vascular lesions in human diabetic retinopathy.Exp Eye Res 2016;146:103-106
7 孙国玲,李筱荣,孙靖.环氧化酶-2在早期糖尿病大鼠视网膜中表达的研究.天津医科大学学报2008;14(3):335-338
8 EL-Asrar AM,Missotten L,Geboes K.Expression of cyelooxygenase-2and downstream enzymes in diabetic fibrovascular epiretina I membranes.Br J Ophthalmol 2008;92(11):1534-1539
9 Schoenberger SD,Kim SJ,Sheng J,et al.Increased prostaglandin E2(PGE2)levels in proliferative diabetic retinopathy.and correlation with VEGF and inflammatory eytokines.Invest Ophthalmol Vis SCi 2012;53(3):5906-5911
10 Tien T,Muto T,Barrette K,et al.Downregulation of Connexin 43promotes vascular cell loss and excess permeability associated with the development of vascular lesions in the diabetic retina.Mol Vis 2014;20(12):732-741
11 Li K,Yao J,Shi L,et al.Reciprocal regulation between proinflammatory cytokine-induced inducible NO synthase(iN OS)and connexin43 in bladder smooth muscle cells.J Biol Chem 2011;286(48):41552-41562
12 Tsuchida S,Arai Y,Kishida T,et al.Silencing the expression of connexin 43 decreases inflammation and joint destruction in experimental arthritis.J Orthop Res 2013;31(4):525-530