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[豚鼠实验性形觉剥夺性和离焦性近视中视蛋白表达的变化]

[Changes of opsin expression in experimental form-deprivation and defocus myopia in guinea pig].

作者信息

Hu Min, Liu Rui, Liu Yong-song, Qian Yi-shan, Chen Chong-da, Yuan Yuan-sheng, Jiang Chun-guang, Chu Ren-yuan

机构信息

Department of Ophthalmology, the Second People's Hospital of Yunnan Province, Kunming, China.

出版信息

Zhonghua Yan Ke Za Zhi. 2012 Feb;48(2):148-52.

Abstract

OBJECTIVE

To investigate the opsin expression in form-deprived and defocus myopia in guinea pig and to study the relationship between the opsin expression and the experimental myopia.

METHODS

Fifty guinea pigs were randomized into form-deprived group, defocus group (n = 20 in each group) and normal group (n = 10). Guinea pigs in form-deprived group wore a diffuser RGP on one eye since one week after birth. Those in defocus group wore -4.00 D RGP on one eye. The contralateral eyes wore 0 D RGP were used as the control. Refraction, axial length and depth of vitreous cavity were measured after 1 and 2 weeks. Animals were sacrificed and the retina were dissected at 10:00 - 12:00 AM. The level of opsin and its mRNA were measured by Western-blot and real-time PCR, respectively.

RESULTS

Two weeks after the experiment, the refraction in form-deprived group and defocus group were (-4.00 ± 0.87) and (-2.00 ± 1.17) D respectively, which were significant different compared with contralateral eyes or normal control group (F = 203.98, 88.66, P < 0.05). These also accompany with increase of axial length and depth of vitreous cavity in form-deprived group and defocus group. Expression of S-opsin mRNA were increased both in form-deprived and defocus groups and the ratios of S-opsin mRNA/β-actin expression were 0.752 ± 0.05 and 1.117 ± 0.13 in two weeks treatment, which were significant different from contralateral eyes (the ratios of S-opsin mRNA/β-actin expression were 0.536 ± 0.04 and 0.772 ± 0.10.t = 6.10, 6.28, P < 0.05). Similar findings were also demonstrated in the expression of L-opsin mRNA, which were increased in form-deprived group and defocus group (the ratios of L-opsin mRNA/β-actin expression were 0.42 ± 0.01) compared with contralateral eyes (the ratios were 0.24 ± 0.0 and 0.34 ± 0.04. t = 6.30, 4.93, P < 0.05) after two weeks experiment. The western-blot results also indicated the high expression of S-opsin and L-opsin level compared to contralateral eyes in form-deprived and defocus groups.

CONCLUSIONS

Cone might be the detector receiving the signal of form-deprivation and defocus. Changes of opsin expression might play a role in the occurrence of experimental myopia in guinea pig.

摘要

目的

研究豚鼠形觉剥夺性近视和离焦性近视中视蛋白的表达情况,探讨视蛋白表达与实验性近视的关系。

方法

将50只豚鼠随机分为形觉剥夺组、离焦组(每组20只)和正常组(10只)。形觉剥夺组豚鼠自出生后1周起,单眼佩戴漫射式硬性透氧性角膜接触镜(RGP);离焦组豚鼠单眼佩戴-4.00 D的RGP。对侧眼佩戴0 D的RGP作为对照。分别于1周和2周后测量屈光、眼轴长度和玻璃体腔深度。上午10:00 - 12:00处死动物,取出视网膜。分别采用蛋白质免疫印迹法(Western-blot)和实时荧光定量聚合酶链反应(real-time PCR)检测视蛋白及其mRNA水平。

结果

实验2周后,形觉剥夺组和离焦组的屈光分别为(-4.00±0.87)D和(-2.00±1.17)D,与对侧眼或正常对照组相比差异有统计学意义(F = 203.98, 88.66, P < 0.05)。形觉剥夺组和离焦组的眼轴长度和玻璃体腔深度也增加。形觉剥夺组和离焦组S-视蛋白mRNA表达均增加,2周处理后S-视蛋白mRNA/β-肌动蛋白表达比值分别为0.752±0.05和1.117±0.13,与对侧眼相比差异有统计学意义(S-视蛋白mRNA/β-肌动蛋白表达比值分别为0.536±0.04和0.772±0.10,t = 6.10, 6.28, P < 0.05)。L-视蛋白mRNA表达也有类似结果,实验2周后,形觉剥夺组和离焦组L-视蛋白mRNA表达增加(L-视蛋白mRNA/β-肌动蛋白表达比值分别为0.42±0.01),与对侧眼相比差异有统计学意义(比值分别为0.24±0.0和0.34±0.04,t = 6.30, 4.93, P < 0.05)。蛋白质免疫印迹结果也显示,形觉剥夺组和离焦组S-视蛋白和L-视蛋白水平高于对侧眼。

结论

视锥细胞可能是接受形觉剥夺和离焦信号的感受器。视蛋白表达变化可能在豚鼠实验性近视的发生中起作用。

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