转醛醇酶与大鼠脉络膜新生血管形成的相关性及其作用机制  

Correlation between transaldolase and choroidal neovascularization and its mechanism in rat models

作　　者：覃珂 许燕[1] 周希瑗[1] 刘丹宁[1] QIN Ke;XU Yan;ZHOU Xiyuan;LIU Danning(Department of Ophthalmology,Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆医科大学附属第二医院眼科,重庆400010

出　　处：《眼科学报》2020年第6期393-399,共7页Eye Science

基　　金：重庆市科学技术委员会(cstc2016jcyjA0122);重庆市卫生和计划生育委员会(ZY201702079)。

摘　　要：目的:探讨转醛醇酶(t ransaldolase,TAL)在氪激光诱导的BN大鼠脉络膜新生血管(choroida l neovascularization,CNV)中的表达变化及其可能的作用机制。方法:给予2组20只BN大鼠行氪激光光凝视网膜,诱导BN大鼠建立CNV模型。在光凝后21 d行荧光素眼底血管造影(fluorescein fundus angiography,FFA),确定模型建立成功后摘除眼球。采用蛋白质印迹法检测TAL、还原型烟酰胺腺嘌呤二核苷酸磷酸(nicotinamide adenine dinucleotide phosphate,NADPH)和还原型谷胱甘肽(glutathione,GSH)蛋白质表达,并测定TAL活性。采用小干扰RNA(small interfering RNA,siRNA)抑制下TAL在人视网膜色素上皮(retinal pigment epithelium,RPE)ARPE-19细胞中的表达,蛋白质印迹法检测抑制前后TAL,NADPH和GSH表达水平的变化。结果:光凝后21 d见明显新生血管生成,表明模型成功建立。CNV增殖区TAL活性明显高于正常大鼠中TAL活性(67.16±3.38 vs 182.57±1.83,P<0.001);TAL蛋白质表达明显高于正常大鼠中TAL表达(0.87±0.33 vs 2.09±0.21,P=0.005);NADPH和GSH表达明显低于其在正常大鼠中的表达(0.92±0.20 vs 0.14±0.05,P<0.01;0.84±0.31 v s 0.29±0.11,P<0.05)。通过对人视网膜色素上皮ARPE-19细胞不做干预、加入LipofectamineTM 2000稀释液、加入TAL-siRNA/LipofectamineTM 2000复合物,将其分为正常对照组、阴性对照组及TAL-siRNA组,干预48 h后,TAL-siRNA组中TAL蛋白质表达明显低于正常对照组和阴性对照组(0.26±0.13 vs 1.39±0.42和1.15±0.19,P<0.01);NADPH和GSH表达明显高于正常对照组(P<0.01)和阴性对照组(P<0.05)。结论:CNV形成的过程中,TAL的活性增强、表达上调,证实TAL与CNV发生发展密切相关,其可能通过调控NADPH和GSH的表达参与CNV的发生发展。Objective:To observe the expression of transaldolase(TAL)in choroidal neovascularization(CNV)induced by krypton laser in BN rats and investigate its possible mechanism.Methods:Twenty rats in two groups were given with krypton laser photocoagulation retina to induce CNV BN rat models.After fluorescein fundus angiography(FFA)was performed 21 days after photocoagulation,the eyeballs were removed,and TAL,nicotinamide adenine dinucleotide phosphate(NADPH)and reduced glutathione(GSH)proteins were detected by Western blot,and TAL activity was determined.The siRNA was used to inhibit the expression of TAL in ARPE-19 cells.The changes of TAL,NADPH and GSH expression levels before and after inhibition were compared by Western blot.Results:Significant neovascularization was observed 21 days after photocoagulation.The TAL activity in the CNV group was significantly higher than that in the control group(67.16±3.38 vs 182.57±1.83,P<0.001);TAL protein expression in the CNV group was significantly higher than that in the control group(0.87±0.33 vs 2.09±0.21,P=0.005);NADPH and GSH expression in the CNV group was significantly lower than that in the control group(0.92±0.20 vs 0.14±0.05,P<0.01;0.84±0.31 vs 0.29±0.11,P<0.05).After TAL-siRNA transfection of ARPE-19 cells for 48h,the expression of TAL protein in the CNV group was significantly lower than that of the control group and negative control group(0.26±0.13 vs 1.39±0.42 vs 1.15±0.19,P<0.01).The expression levels of NADPH and GSH in the CNV group were significantly higher than those in the control group(P<0.01)and negative control group(P<0.05).Conclusion:CNV is positively correlated with TAL expression,and TAL can participate in the development of CNV by regulating the expression levels of NADPH and GSH.

关 键 词：转醛醇酶 脉络膜新生血管 磷酸戊糖途径 

分 类 号：R773.4[医药卫生—眼科]