摘要
目的:通过球旁注射单胺囊泡转运体2(VMAT2)非竞争性抑制剂丁苯那嗪(TBZ)探讨胞外多巴胺水平下调对三色豚鼠屈光发育以及近视的影响。方法:实验研究。152只正常3周龄豚鼠,随机分为正常视觉环境(NV)组和形觉剥夺(FD,右眼单眼剥夺)组。NV组又随机分为空白对照组(NC,n=18),溶剂对照组[注射二甲基亚枫(DMSO),NV+DMSO,n=18)],低剂量TBZ组(NV+30 ng,n=22),高剂量TBZ组(NV+300 ng,n=20)。FD组又随机分成单纯FD组(FD,n=17),FD+溶剂对照组(FD+DMSO,n=18),FD+低剂量TBZ组(FD+30 ng,n=21),FD+高剂量TBZ组(FD+300 ng,n=18)。溶剂注射组和TBZ给药组每天右眼球旁注射0.1%DMSO或者相应剂量TBZ 100μl。实验前、实验后2周和4周分别检测屈光度和眼轴相关参数。实验4周后采用过量10%水合氯醛处死豚鼠,免疫荧光染色检测多巴胺受体下游信号分子p-CREB的变化。数据采用独立样本t检验、单因素方差分析进行分析。结果:在正常视觉环境下,TBZ注射2周可诱导豚鼠产生近视,NV+DMSO、NV+30 ng和NV+300 ng组双眼屈光度差值(右眼-左眼)分别为(0.04±0.31)D、(-0.90±0.84)D和(-0.75±0.66)D(F=11.133,P<0.001),同时伴玻璃体腔深度加深,3组双眼玻璃体腔深度差值分别为(-0.01±0.03)mm、(0.02±0.03)mm和(0.02±0.04)mm(F=6.001,P=0.004)。注射TBZ 4周豚鼠的近视量没有进一步加深,3组豚鼠双眼屈光度的差值分别为(-0.06±0.41)D、(-1.14±1.61)D和(-0.84±1.41)D(F=3.569,P=0.035)。正常屈光发育的豚鼠注射TBZ 4周后,神经节细胞层和内核层的p-CREB表达下调(t=4.825,P=0.003;t=8.109,P<0.001)。TBZ注射对FD豚鼠2周和4周时的屈光和眼轴都没有影响。结论:通过TBZ抑制单胺囊泡转运体功能可诱导正常豚鼠出现近视,但对形觉剥夺性近视没有显著影响。单胺囊泡转运体功能受抑制可能通过使单胺类递质的释放减少,引起相应递质,特别是多巴胺水平的降低进而诱导出近视。
Objective:To investigate the effect of reduced extracellular dopamine(DA)levels on refractive development in guinea pigs by periocularly injecting a noncompetitive vesicular monoamine transporter 2(VMAT2)antagonist,tetrabenazine(TBZ).Methods:Three-week-old guinea pigs were randomly divided into 8 groups in this experimental group.Seventy-eight animals were raised under normal vision(NV)and divided into the following groups:the normal control(NC,n=18),vehicle(dimethyl sulfoxide,NV+DMSO,n=18),low-dose TBZ(NV+30 ng,n=22),and high-dose TBZ(NV+300ng,n=20)groups.Seventy-four animals raised with form deprivation(FD)were divided into the following groups:FD(n=17),FD+DMSO(n=18),FD+30 ng(n=21)and FD+300 ng(n=18)groups.DMSO,TBZ(30 ng or 300 ng)was periocularly injected into the right eyes daily in 100μl of 0.1%DMSO.Refractive changes were evaluated prior to treatment and 2 weeks and 4 weeks after treatments.p-CREB immunofluorescent staining evaluated DA receptor activation.Data were analyzed using one-way ANOVA and an independent samples t-test.Results:After 2 weeks of TBZ injections,guinea pigs developed myopia in the NV environment.The inter-ocular difference in refraction for NV+DMSO,NV+30 ng and NV+300 ng were 0.04±0.31 D,-0.90±0.84 D and-0.75±0.66 D,respectively(F=11.133,P<0.001).The vitreous chamber depth in these three groups was increased consistently with the inter-ocular differences of-0.01±0.03 mm,0.02±0.03 mm and 0.02±0.04 mm(F=6.001,P=0.004).Four weeks of TBZ injection could not further increase myopia.The inter-ocular differences between NV+DMSO,NV+30 ng and NV+300 ng were-0.06±0.41 D,-1.14±1.61 D and-0.84±1.41 D,respectively(F=3.569,P=0.035).Four weeks of TBZ application significantly reduced the number of cells that expressed p-CREB in the retinal ganglion cell layer and inner nuclear layer(t=4.825,P=0.003,t=8.109,P<0.001).TBZ injections did not affect the development of form deprivation myopia(FDM)in guinea pigs.Conclusions:The VMAT2 function inhibition by TBZ induces myopia in guinea pigs raised
作者
祝瑞雪
裴晓梦
胡洋
支知娜
张森
周翔天
瞿佳
Ruixue Zhu;Xiaomeng Pei;Yang Hu;Zhina Zhi;Sen Zhang;Xiangtian Zhou;Jia Qu(Eye Hospital,Wenzhou Medical University,Wenzhou 325027,China)
出处
《中华眼视光学与视觉科学杂志》
CAS
CSCD
2018年第6期326-332,共7页
Chinese Journal Of Optometry Ophthalmology And Visual Science
基金
国家自然科学青年基金(80214057)
国家自然科学基金(81470659、80215089)
国家自然科学基金优秀青年科学基金(81422007)
浙江省自然科学基金重点项目(LZ14H120001)
浙江省自然科学基金面上项目(LY18H120006)
关键词
近视
胞外多巴胺
单胺囊泡转运体2
豚鼠
丁苯那嗪
myopia
extracellular dopamine
vesicular monoamine transporter 2
guinea pig
tetrabenazine
