CAJ | 학술논문

目的探讨辅助性T细胞(Th细胞)在多聚物-1( Cop-1)对高眼压大鼠视网膜神经节细胞保护中可能发挥的作用.方法 Wistar大鼠分别结扎3根巩膜上静脉,建立高眼压模型96只,完全随机法分入Cop-1免疫组48只,磷酸盐缓冲液(PBS)免疫组48只,另10只正常大鼠作对照.于免疫后3、7、10、17、24、31 d,用免疫荧光染色法观察视网膜切片中Th1、Th2数量变化,并通过免疫蛋白印迹法检测不同组别大鼠视网膜中IL-4蛋白的变化.结果免疫荧光显示,免疫后7d,Cop-1组与PBS组视网膜上Th1数量增多,分别为(216±21 )/mm2与(194±27 )/mm2,此后两组Th1数量随时间延长而减少,在各个时间点均差异无统计学意义;免疫后7d,Cop-1组视网膜上Th2数量远大于PBS组,差异有统计学意义[(300±28)/mm2和( 129±27)/mm2,P＜O.01],随后Cop-1组的rh2数量随时间延长而减少,仍大于PBS组.蛋白印迹表明,免疫后3d,Cop-1组视网膜上IL-4相对蛋白含量已高于PBS组;7 d,Cop-1组IL-4含量为2.11±0.06,高于PBS组0.57±0.05,随后,每个时间点Cop-1组的IL-4含量均高于PBS组(均P＜0.05).结论 Cop-1引起高眼压大鼠视网膜中Th2型细胞因子IL-4聚集,提示Cop-1对视网膜神经节细胞保护可能是通过特异性Th2细胞发挥效应.
목적 탐토보조성T세포(Th세포)재다취물-1( Cop-1)대고안압대서시망막신경절세포보호중가능발휘적작용.방법 Wistar대서분별결찰3근공막상정맥,건립고안압모형96지,완전수궤법분입Cop-1면역조48지,린산염완충액(PBS)면역조48지,령10지정상대서작대조.우면역후3、7、10、17、24、31 d,용면역형광염색법관찰시망막절편중Th1、Th2수량변화,병통과면역단백인적법검측불동조별대서시망막중IL-4단백적변화.결과 면역형광현시,면역후7d,Cop-1조여PBS조시망막상Th1수량증다,분별위(216±21 )/mm2여(194±27 )/mm2,차후량조Th1수량수시간연장이감소,재각개시간점균차이무통계학의의;면역후7d,Cop-1조시망막상Th2수량원대우PBS조,차이유통계학의의[(300±28)/mm2화( 129±27)/mm2,P＜O.01],수후Cop-1조적rh2수량수시간연장이감소,잉대우PBS조.단백인적표명,면역후3d,Cop-1조시망막상IL-4상대단백함량이고우PBS조;7 d,Cop-1조IL-4함량위2.11±0.06,고우PBS조0.57±0.05,수후,매개시간점Cop-1조적IL-4함량균고우PBS조(균P＜0.05).결론 Cop-1인기고안압대서시망막중Th2형세포인자IL-4취집,제시Cop-1대시망막신경절세포보호가능시통과특이성Th2세포발휘효응.
Objective To explore the morphological and expressional changes of Th1 cells and Th2 cells in retina of a rat model of glaucoma vaccinated by Cop-1 ( Copolymer-1 ) and elucidate the possible neuroprotection roles played by Th1/Th2.Methods After modeling,the aqueous outflow from the right eyes was blocked by a ligation of three of four episcleral veins.There were 48 rats with elevated IOP ( intraocular pressure) immunized by Cop-1 ( Cop-1 group),48 rats with elevated IOP immunized by PBS ( phosphate buffered saline) (PBS group) and 10 rats without any treatment (normal group).The experimental rats were immunized with Cop-1/PBS emulsified in a total volume of 0.4 ml complete Freund's adjuvant.The immunization was administered subcutaneously at the base of tail.Immunofluorescence was employed to test the distribution and activation of Thl and Th2 cells in retina at Days 3,7,10,17,24 and 31 post immunization respectively for each group.Western blot was selectively performed according to the results of immunofluorescence to verify if there was a similar variation of the retinal expression of IL-4 protein.Results The results of immunofluorescence showed the numbers of Thl cells peaked at Day 7 in both Cop-1 ( (216 ± 21 )/mm2) and PBS groups (( 194 ±27 )/mm2 ).And no statistical significance existed between two groups (P＞0.05).The numbers of Th2 cells in the experimental groups peaked at Day 7 with statistical significance ( Cop-1 group:300 ± 28/mm2 vs PBS group:129 ± 27/mm2 ) ( P ＜ 0.01 ).With the prolongation of experimental period,the number of Th2 cells decreased gradually in the Cop-1 group but remained greater than that of the PBS group afterward ( P ＜ 0.05 ).The Western blot results showed that the expression of IL-4 in the Cop-1 group ( 1.91 ± 0.05 ) was significantly higher than that of the PBS group (0.51 ±0.04) from Day 3 and peaked at Day 7 (2.11 ±0.06 vs 0.57 ±0.05).Then the IL-4 expression decreased gradually in the COP-1 group but still represented statistical significance versus the PBS group until Day 31 post-immunization ( P ＜0.001 ).Conclusion The retinal activation and accumulation of IL-4 are found in a rat model of chronic glaucoma immunized by Cop-1.Thus Th2 cells may play vital roles in the Cop-l-induced neuroprotective autoimmune responses.