妥拉唑啉控制雏鸡实验性近视进展的有效性和安全性

Effectiveness and Safety of Tolazoline in the Experimental Myopia Control in Chicks

目的::研究肾上腺素α受体阻滞剂妥拉唑啉控制雏鸡实验性近视的有效性和安全性,并探讨其作用机制。方法::实验研究。于2021年3月将61只6日龄白莱航鸡随机分为形觉剥夺对照组(FDM)、形觉剥夺后妥拉唑啉组(FDM+T100)、形觉剥夺后妥拉唑啉联合L-NAME组(FDM+T100+L-NAME)和离焦诱导对照组(LIM)、离焦诱导后妥拉唑啉组(LIM+T100)、离焦诱导后妥拉唑啉联合L-NAME组(LIM+T100+L-NAME)进行有效性研究;空白对照组(Blank)、单纯妥拉唑啉干预组(Blank+T100)进行安全性研究。所有雏鸡均取右眼进行实验干预,妥拉唑啉干预组在造模的同时每天进行玻璃体腔注射12.5μL妥拉唑啉,妥拉唑啉联合L-NAME干预组注射等量妥拉唑啉和L-NAME,对照组注射等体积0.9%氯化钠溶液,持续6 d。记录和统计分析各组处理前后的屈光度和眼球生物参数。安全性研究中,对Blank组和Blank+T100组雏鸡诱导结束后进行视网膜电图检查。雏鸡处死后眼球固定切片进行TUNEL染色观察视网膜功能损害程度。不同近视模型中对照组与T100组、T100组与T100+L-NAME组两两比较,采用独立样本t检验和Mann-Whitney U检验。结果::经过6 d处理后,在形觉剥夺模型中,与FDM组相比,FDM+T100组的屈光度近视化减少(t=-16.16,P<0.001),眼轴增长量减少(t=12.41,P<0.001),脉络膜厚度减薄量减少(t=-2.46,P=0.031);与FDM+T100组相比,FDM+T100+L-NAME组屈光度近视化增加(t=4.20,P=0.001),眼轴增长量增加(t=-2.33,P=0.033)。在离焦诱导模型中,与LIM组相比,LIM+T100组屈光度近视化减少(t=-5.77,P<0.001),眼轴增长量减少(t=4.19,P<0.001);与LIM+T100组相比,LIM+T100联合L-NAME组屈光度近视化增加(t=3.74,P=0.001),眼轴增长量增加(t=-3.20,P=0.005)。安全性研究中,Blank组和Blank+T100组的屈光度及眼球生物参数变化值之间差异均无统计学意义(均P>0.05);ERG结果显示,Blank组和Blank+T100组相比,双眼各检测指标比值之间差异均无统计学意义(均P>0.05)。TUNEL染色显示各组均未见显著凋亡。结论::妥拉唑啉能够有效延缓雏鸡实验性近视的屈光度和眼轴进展,但在应用L-NAME后减弱,提示在妥拉唑啉扩张脉络膜血管时可能也有一氧化氮共同参与发挥作用。

Objective:To investigate the efficacy and safety of the adrenergicα-receptor blocker tolazoline in the treatment of experimental myopia in chicks,and explore its mechanism.Methods:In this experimental study,totally 616-day-old White Lehorn chickens were randomly divided into form deprivation control group(FDM),tolazoline treatment group after form deprivation(FDM+T100),tolazoline combined with NOS inhibitor L-NAME group after form deprivation(FDM+T100+L-NAME),lens induction control group(LIM),tolazoline treatment group after lens induction(LIM+T100),and tolazoline combined with NOS inhibitor L-NAME group after lens induction(LIM+T100+L-NAME)for efficacy study.Chickens were also divided into blank control group(Blank)and tolazoline treatment group(Blank+T100)for safety study.The experimental intervention was performed on the right eyes of all chicks.Eyes requiring tolazoline(T100)treatment were injected with 12.5μL tolazoline,eyes requiring tolazoline combined L-NAME(T100+L-NAME)were injected with equal equivalent tolazoline and L-NAME,and the control group was injected with normal saline for 6 days.The diopter and eye biological parameters of each group before and after treatment were recorded and analyzed statistically.In the safety study,ERG examinations were performed on blank group and blank+T100 group.After sacrificing the chicks,the eyeballs were fixed and sliced for TUNEL staining to observe retinal damage.The experimental group and the control group in different myopia model were compared in pairs in this study.The independent samples t-test was used for diopter,vitreous depth,choroid thickness,and axial length which satisfy the normal distribution,and the Mann-Whitney U test was used for the electroretinogram ratio results which did not meet the normal distribution.Results:After 6 days of treatment,in the form deprivation model,the FDM+T100 group had reduced refractive myopia(t=-16.16,P<0.001),reduced amount of axial length growth(t=12.41,P<0.001),and reduced amount of choroidal thickness thinning(t=-2.46,P=0.031)compared to the FDM group.Compared to the FDM+T100 group,the FDM+T100+L-NAME group had increased refractive myopia(t=4.20,P=0.001),increased amount of axial length growth(t=-2.33,P=0.033).In the lens induction model,when compared with the LIM group,the refractive diopter was reduced in the LIM+T100 group(t=-5.77,P<0.001),axial length growth was reduced(t=4.19,P<0.001).When compared with the LIM+T100 group,the refractive diopter was increased in the LIM+T100+L-NAME group(t=3.74,P=0.001),and increased amount of axial length growth(t=-3.20,P=0.005).In the safety study,there was no significant difference in the diopter,the changes of eyeball biological parameters and ERG right/left eye ratio between the blank control group and the simple tolazoline treatment group.No significant apoptosis was found among the groups in TUNEL staining.Conclusions:Tolazoline can effectively delay the diopter and axial progression of experimental myopia in chicks.But this effect was weakened by NOS inhibitor L-NAME.It is suggested that nitric oxide(NO)may also play an important role in the dilation of choroidal blood vessels by tolazoline.