CAJ | 학술논문

目的建立豚鼠形觉剥夺性近视动物模型,玻璃体腔内注射不同浓度的N-甲基-D-天冬氨酸受体1(NMDAR1)反义寡核苷酸(ASON),观察NMDAR1 ASON对豚鼠近视的调节,探讨其在近视发病机制中的作用.方法实验研究.3周龄三色豚鼠随机分为6组:A组(正常对照,10只)、B组(右眼遮盖3周,10只)、C1组(右眼遮盖3周+玻璃体腔注射100 ng NMDAR1正义寡核苷酸)、C2组(右眼遮盖3周+玻璃体腔注射生理盐水)、C3组(右眼遮盖3周+玻璃体腔注射10 ng NMDAR1 ASON)、C4组(右眼遮盖3周+玻璃体腔注射100 ng NMDAR1 ASON).C1～C4组在遮盖2周时予以玻璃体腔注药1周.实验前及实验3周时对各组进行视网膜检影和A超测眼轴,免疫组化法测NMDAR1的蛋白表达变化,RT-PCR定量检测NMDAR1的mRNA含量变化.各组测量值之间的差异采用one-way ANOVA检验,将各组测量值与NMDAR1 ASON注射浓度进行二元线性相关分析.结果 A、B、C1～C4组之间屈光度、眼轴长度、NMDAR1蛋白表达及mRNA含量差异有统计学意义(F=55.247、142.213、43.215、592.435,P＜0.05).B组及所有C组右眼呈近视状态,眼轴较自身对侧眼长.B组与C1、C2组遮盖眼屈光状态及眼轴、NMDAR1的蛋白表达及mRNA含量变化表达差异无统计学意义.C2、C3、C4组遮盖眼依次近视屈光度数下降,眼轴延长减慢,NMDAR1的蛋白含量和mRNA含量下调.屈光度与注射浓度呈正相关(r=-0.695,P＜0.05),眼轴长度、NMDAR1蛋白表达、NMDAR1 mRNA表达与其呈负相关(r=-0.731、-0.625、-0.821,P＜0.05).结论形觉剥夺可明显上调豚鼠视网膜NMDAR1的蛋白及mRNA的表达,NMDAR1 ASON玻璃体腔注射能通过下调NMDAR1的蛋白含量及mRNA表达,来减缓近视的进展.
목적 건립돈서형각박탈성근시동물모형,파리체강내주사불동농도적N-갑기-D-천동안산수체1(NMDAR1)반의과핵감산(ASON),관찰NMDAR1 ASON대돈서근시적조절,탐토기재근시발병궤제중적작용.방법 실험연구.3주령삼색돈서수궤분위6조:A조(정상대조,10지)、B조(우안차개3주,10지)、C1조(우안차개3주+파리체강주사100 ng NMDAR1정의과핵감산)、C2조(우안차개3주+파리체강주사생리염수)、C3조(우안차개3주+파리체강주사10 ng NMDAR1 ASON)、C4조(우안차개3주+파리체강주사100 ng NMDAR1 ASON).C1～C4조재차개2주시여이파리체강주약1주.실험전급실험3주시대각조진행시망막검영화A초측안축,면역조화법측NMDAR1적단백표체변화,RT-PCR정량검측NMDAR1적mRNA함량변화.각조측량치지간적차이채용one-way ANOVA검험,장각조측량치여NMDAR1 ASON주사농도진행이원선성상관분석.결과 A、B、C1～C4조지간굴광도、안축장도、NMDAR1단백표체급mRNA함량차이유통계학의의(F=55.247、142.213、43.215、592.435,P＜0.05).B조급소유C조우안정근시상태,안축교자신대측안장.B조여C1、C2조차개안굴광상태급안축、NMDAR1적단백표체급mRNA함량변화표체차이무통계학의의.C2、C3、C4조차개안의차근시굴광도수하강,안축연장감만,NMDAR1적단백함량화mRNA함량하조.굴광도여주사농도정정상관(r=-0.695,P＜0.05),안축장도、NMDAR1단백표체、NMDAR1 mRNA표체여기정부상관(r=-0.731、-0.625、-0.821,P＜0.05).결론 형각박탈가명현상조돈서시망막NMDAR1적단백급mRNA적표체,NMDAR1 ASON파리체강주사능통과하조NMDAR1적단백함량급mRNA표체,래감완근시적진전.
Objective To explore the possible mechanisms of myopia.Form-deprivation myopia was induced in guinea pigs and N-methyl-D-aspartate receptor 1 (NMDAR1) antisense oligonucleotides were injected intravitreously.Methods In this experimental study,3-week-old guinea pigs were divided into six groups (each n=10):group A was the control group; group B underwent monocular form-deprivation (MD) for 3 weeks; group C1 received intravitreous injection of 100 ng NMDAR1 sense oligonucleotides to MD eyes; group C2 received intravitreous injection of saline to MD eyes;group C3 received intravitreous injection of 10 ng NMDAR1 antisense oligonucleotides to MD eyes;group C4 received intravitreous injection of 100 ng NMDAR1 antisense oligonucleotides to MD eyes.Groups C1～C4 received the intravitreous injection at the second week of deprivation.Refraction and axial length of the eyes were measured and the expression of NMDAR1 was assayed by immunohistochemistry and RT-PCR before the test and 3 weeks later.Differences between groups were assessed using one-way ANOVA,and the correlation between NMDAR1 antisense oligonucleotides and refraction,axial length were analyzed with linear correlation.Results There were statistical differences in the degree of myopia,axial length and level of NMDAR1 protein and mRNA among groups A,B,C1～C4 (F=55.247,142.213,43.215,592.435,P＜0.05).In group B and groups C1～C4,myopia was more severe and eye axial length was longer in MD eyes.There were no significant differences in the degree of myopia,axial length or the level of NMDAR1 protein and mRNA among groups B,C1 and C2.The degree of myopia was gradually less severe,axial length was gradually shortened and the level of NMDAR1 was gradually down regulated among groups C2,C3 and C4 and the effect was concentration-dependent.The degree of myopia had a positive correlation to the concentration gradient (r=-0.695,P＜0.05) while axial length and the level of NMDAR1 expression had a negative correlation (r=-0.731,-0.625,-0.821,P＜0.05).Conclusion Significant up-regulation can be observed in the expression of NMDAR1 in the level of mRNA and protein in MD eyes.With NMDAR1 antisense oligonucleotides injected intravitreously,the expression of NMDAR1 is down-regulated along with reduced development of myopia.