双调蛋白抗体对双眼诱导型近视豚鼠眼部的影响

摘要

目的 探讨双调蛋白抗体对双眼诱导型近视豚鼠眼轴长度、屈光度及后极部巩膜厚度等的影响.方法 60只三色短毛豚鼠随机分为5组:正常对照组、近视模型组、低剂量组、中剂量组、高剂量组,每组12只.豚鼠双眼戴-10 D透镜诱导近视:正常组不作任何处理,戴镜2周后,低、中、高剂量组豚鼠右眼玻璃体内注射低、中、高剂量双调蛋白抗体,同时左眼玻璃体内注射同等剂量的乳酸钠林格注射液(buffer组),注射抗体前后持续戴镜.于戴镜前、戴镜后2周、注射3次抗体后测量豚鼠的屈光度和眼轴长度.注射3次抗体后,过碘酸雪夫染色法检测豚鼠眼球后极部视网膜、脉络膜及巩膜的厚度,实时荧光定量PCR和免疫荧光法检测视网膜中双调蛋白及表皮生长因子受体mRNA、蛋白的表达.结果 透镜诱导2周后,近视模型组与正常对照组比较眼轴长度增长、屈光度缩短,差异均有统计学意义(均为P＜0.01).注射3次抗体后,与buffer组相比,低、中、高剂量组眼轴长度均缩短,屈光度均上升,后极部巩膜厚度均明显增加,差异均有统计学意义(均为P ＜0.05).各组后极部视网膜厚度无明显变化.实时荧光定量PCR结果和免疫荧光检测结果显示近视模型组及buffer组视网膜中双调蛋白mRNA和蛋白表达增加,高、中、低剂量组视网膜中双调蛋白mRNA和蛋白表达均下降,与buffer组相比,低剂量组视网膜中表皮生长因子受体表达下降(t=2.606,P=0.022).结论 在近视造模豚鼠的玻璃体内注射双调蛋白抗体后,豚鼠屈光度上升、眼轴长度明显缩短、后极部巩膜厚度增加,其机制可能与双调蛋白表达的减少有关.%Objective To investigate the effects of amphiregulin antibody on the axial length,diopter and posterior sclera thickness in eyes of bilateral lens-induced myopic guinea pigs.Methods A total of 60 healthy three-colored short-hair guinea pigs were randomly divided into 5 groups:myopic model group,low dose group,medium dose group,high dose group and normal control group,12 rats in each group,and both eyes of the guinea pigs in the former four groups were induced by-10 D lens,while the normal group did not make any treatment.After wearing of goggles for 2 weeks,the right eyes of guinea pigs were intravitreally administrated with 5 μg,10 μg,20 μg anphiregulin antibody for the low,medium and high dose group,respectively.At the same time,the left eyes of these groups were given the same dose of Ringer' s buffer (buffer group).Continuous wearing of the lenses before and after injection of antibody was allowed.Diopter and axial length of guinea pigs were measured before wearing of the lenses,2 weeks after wearing lenses and 5 weeks after the experiment (after 3-times injections of antibody).Moreover,the thicknesses of retina,choroid and sclera in the posterior pole were detected by PAS staining.Finally,the expression of amphiregulin and EGFR mRNA and protein was detected by real time quantitative PCR and immunofluorescence.Results After 2 weeks of lens induction,the axial length of the myopic model group increased,but the diopter decreased when compared with the normal control group,and the differences were statistically significant (both P ＜ 0.05).Compared with the buffer group,after intravitreal injection of amphiregulin antibody,the axial length in the low,medium,and high dose groups decreased,while the diopter increased,and the scleral thickness at the posterior pole increased significantly in a dose-dependent manner,with statistically significant differences (all P ＜ 0.05).However,there was no significant change in retinal thickness at the posterior pole of all groups.Real-time quantitative PCR and immunofluorescence showed the expression of amphiregulin mRNA and protein in the retina was upregulated the myopic model group and buffer group,but downregulated in the high,medium,and low dose groups.Furthermore,when compared with the buffer group,the expression level of epidermal growth factor receptor in the retina of the low dose group was decreased (t =2.606,P =0.022),but there was no significant difference in the other groups.Conclusion After injection of amphiregulin antibody into the eyes of bilateral lens-induced myopic guinea pigs,the diopter increases,but the axial length is significantly shortened,and the posterior sclera is thickened,which may involve a decrease in the expression of amphiregulin.