形觉剥夺性近视眼巩膜组织中M受体5种亚型表达的变化

Changes of 5 muscarinic receptor subtypes in scleral tissues of eyes with form deprivation myopia

目的探讨豚鼠形觉剥夺性近视眼中,巩膜组织中M受体5种亚型的表达变化。方法1～2周龄的三色豚鼠36只,随机分为形觉剥夺性近视眼组(FDM组,21眼)、自身对照组(FC组,21只21眼)和正常组(N组,15只30眼)。FDM组制作动物模型,21d后测量3组动物眼屈光度和眼轴长度。提取各组眼巩膜组织总RNA和蛋白质,半定量RT-PCR和Westernblotting检测M受体5种亚型的表达变化。结果形觉剥夺21d后,FDM组与FC组、N组的屈光度和眼轴之间的差异均有统计学意义(均为P<0.05)。FDM组产生相对近视(-4.28±1.70)D,眼轴相对增长(0.27±0.17)mm。半定量RT-PCR结果显示:后极部巩膜组织中,FDM组与FC组、N组相比较,M1和M4亚型的表达均有显著性增加(均为P<0.05),而M2、M3和M5亚型的表达差异均无统计学意义(均为P>0.05),其中FDM组中M1亚型的表达增加了18.67%,M4亚型增加了26.48%(均为P<0.01)。Westernblotting结果显示:后极部巩膜组织中,FDM组与FC组、N组相比较,M1和M4亚型蛋白表达均有显著增加(均为P<0.01),而M2、M3和M5亚型的表达差异均无统计学意义(均为P>0.05),其中FDM组中M1亚型蛋白表达增加了24.65%,M4亚型蛋白增加了49.11%(均为P<0.01)。结论在豚鼠形觉剥夺性近视的形成中,后极部巩膜组织的M1和M4受体的表达显著性增加,提示在近视眼发生和发展中,巩膜组织的M受体可能参与调控眼球的生长。

Objective To investigate the changes of 5 muscarinic（M）receptor subtypes in scleral tissues of guinea pigs with form deprivation myopia（FDM）.Methods Thirty six tri-color guinea pigs at postnatal 1 week to 2 weeks were randomly divided into FDM group（21 guinea pigs,21 eyes）,self-control group（FC group;21 eyes）and normal control group（N group;15 guinea pigs,30 eyes）.Animal models were made in FDM group,diopter and axial length of guinea pigs in 3 groups were measured at 21 days later.Total RNA and protein of scleral tissue were extracted. Changes of 5 M receptor subtypes were measured by semi-quantitative RT-PCR and Western blotting. sulta At 21 days after form deprivation, there was statistical difference in diopter and axial length in FDM group compared with FC group or N group（ both P 〈0.05 ）. The relative myopia was （ -4.28±1.70） D and axial length increased （0. 27± 0. 17） mm in FDM group. Semi-quantitative RT-PCR showed that there was significant increase of Ml and M4 subtypes at posterior slcera in FDM group compared with FC group or N group （ both P 〈 0.05 ）, but no difference was found in M2, M3 and M5 subtypes （ all P 〉 0.05 ）. The expression of M1 subtypes increased 18.57% and M4 subtypes increased 25.48% in FDM group （both P 〈 0.01 ）. Western blotting showed that there was significant increase of M1 and M4 subtypes at posterior slcera in FDM group compred with FC group or N group （ both P 〈 0.01 ）, but no difference was found in M2, M3 and M5 subtypes （ all P 〉 0.05 ） The expression of M1 subtypes increased 24.55% and M4 subtypes increased 49. 11% in FDM group （ both P 〈 0.01 ）. Conelusion In the process of FDM in guinea pigs,the expression of M1 receptor and M4 receptor at posterior sclera significantly increase, which indicate that M receptor may pay a role in regulating the growth of eyeball at scleral tissue in the occurrence and development of myopia.