驻景丸加减方对形觉剥夺性近视小鼠视网膜厚度及细胞凋亡的影响
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国家自然科学基金项目(No.81674031); 四川省科技计划项目(No.2021YJ0252); 成都中医药大学“杏林学者”项目(No.XSGG2020003)


Effect of modified Zhujing pill on retinal thickness and apoptosis in form deprivation myopia mice
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National Natural Science Foundation of China(No.81674031); Sichuan Science and Technology Program Project(No.2021YJ0252); Chengdu University of Traditional Chinese Medicine(TCM)“Xinglin Scholars” Project(No.XSGG2020003)

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    摘要:

    目的:探讨驻景丸加减方对形觉剥夺性近视小鼠视网膜厚度及细胞凋亡的影响。

    方法:将72只3周龄C57BL/6J小鼠随机分为对照组1、模型组1、干预组1、对照组2、模型组2及干预组2,每组12只小鼠,前三组实验进行3wk,后三组实验进行6wk,除对照组1和对照组2外,所有小鼠均用半透明眼罩遮盖右眼诱导形觉剥夺性近视动物模型,干预组1在造模同时而干预组2则在实验3wk后灌胃驻景丸加减方混悬液0.546g/(kg·d)(0.1mL/d),其余组小鼠灌胃等量生理盐水0.1mL/d,实验前后均测眼轴,实验结束时取小鼠视网膜行HE染色用于观察视网膜各层厚度改变,免疫组织化学(IHC)和western blotting(WB)用于检测Bcl-2和Caspase3蛋白的表达。

    结果:在实验结束时模型组1的眼轴比对照组1显著增加(P<0.01),干预组1的眼轴显著低于模型组1(P<0.01),模型组2的眼轴比对照组2显著增加(P<0.01),干预组2的眼轴明显低于模型组2(P<0.01); 模型组1的NFL和ONL的厚度比对照组1显著变薄(P<0.01),模型组1的INL厚度比对照组1显著变薄(P<0.05),干预组1的NFL、INL和ONL厚度较模型组1显著增加(P<0.05); 模型组2的NFL、INL和ONL的厚度比对照组1显著变薄(P<0.01); IHC检测显示Bcl-2蛋白平均光密度模型组1显著低于对照组1(P<0.05),干预组1显著高于模型组1(P<0.01),模型组2显著低于对照组2(P<0.01),干预组2显著高于模型组2(P<0.01); Caspase3蛋白平均光密度模型组1显著高于对照组1(P<0.01),干预组1显著低于模型组1(P<0.01),模型组2显著高于对照组2(P<0.05),干预组2显著低于模型组2(P<0.01); WB检测证明,Bcl-2的蛋白相对表达水平模型组1显著低于对照组1(P<0.01),干预组1显著高于模型组1(P<0.01),模型组2显著低于对照组2(P<0.01),干预组2显著高于模型组2(P<0.01); Caspase3的蛋白相对表达水平模型组1显著高于对照组1(P<0.01),干预组1显著低于模型组1(P<0.01),干预组2显著低于模型组2(P<0.05)。

    结论:驻景丸加减方可通过调控凋亡相关蛋白Bcl-2和Caspase3的表达,减轻近视形成过程中及已形成近视小鼠视网膜细胞凋亡,从而起到干预形觉剥夺近视小鼠视网膜厚度变薄的作用。

    Abstract:

    AIM: To investigate the effect of modified Zhujing pill on retinal thickness and apoptosis in form deprivation myopia(FDM)mice.

    METHODS: Totally 72 C57BL/6J mice aged 3-week-old were randomly divided into control group 1, model group 1, intervention group 1, control group 2, model group 2 and intervention group 2, with 12 mice in each group. The first three groups were tested for 3wk and the last three groups were tested for 6wk, except for the groups of control 1 and control 2, all the mice used translucent goggles to cover their right eyes for form deprivation. The mice of intervention 1 and intervention 2 were respectively given intragastric administration modified Zhujing pill suspension 0.546g/(kg·d)(0.1mL/d)for 3wk at the beginning and after 3wk of the experiment. Same amount of saline was given to mice in other groups at the same time of modeling. The eye axis was measured before and after the experiment. At the end of the experiment, the eye of mice was taken for HE staining to observe the thickness changes of each layer of retina. Immunohistochemistry(IHC)and western blotting(WB)were used to detect Bcl-2 and Caspase3 expression of protein.

    RESULTS: At the end of the experiment, the axis of model group 1 was significantly higher than that of control group 1(P<0.01), the axis of intervention group 1 was significantly lower than that of model group 1(P<0.01), and the axis of model group 2 was significantly higher than that of control group 2(P<0.01), the eye axis of intervention group 2 was significantly lower than that of model group 2(P<0.01); HE staining showed that the thickness of NFL and ONL of model group 1 was significantly thinner than that of control group 1(P<0.01). The thickness of INL of model group 1 was significantly thinner than that of control group 1(P<0.05), and the thickness of NFL, INL and ONL of intervention group 1 was significantly higher than that of model group 1(P<0.05); The thickness of NFL, INL and ONL model group 2 was significantly thinner than that of control group 1(P<0.01); IHC testing showed that the average optical density of Bcl-2 protein in model group 1 was significantly lower than that of control group 1(P<0.05), which in intervention group 1 was significantly higher than that of model group 1(P<0.01), and which in the average optical density of Bcl-2 protein of model group 2 was significantly lower than that of control group 2(P<0.01), which in intervention group 2 was significantly higher than that of model group 2(P<0.01); Caspase 3 protein average optical density of model group 1 was significantly higher than that of control group 1(P<0.01), which in intervention group 1 was significantly lower than that of model group 1(P<0.01), which in model group 2 was significantly higher than that of control group 2(P<0.05), which in intervention group 2 was significantly lower than that of model group 2(P<0.01); WB test proved that the relative expression level of Bcl-2 protein in model group 1 was significantly lower than that of control group 1(P<0.01), which in intervention group 1 was significantly higher than that of model group 1(P<0.01), and which in model group 2 was significantly lower than that of control group 2(P<0.01), which in intervention group 2 was significantly higher than that of model group 2(P<0.01); The relative expression level of Caspase3 protein in model group 1 was significantly higher than that of control group 1(P<0.01), which in intervention group 1 was significantly lower than that of model group 1(P<0.01), the intervention group 2 was significantly lower than that of model group 2.

    CONCLUSION: The results show that the modified Zhujing pill can interfere with the pathological changes of retinal thickness thinning in the process of myopia and formed myopia mice by regulating the expression of apoptosis-related proteins Bcl-2 and Caspase3, and alleviating the apoptosis of retinal cells in myopia formation and myopia mice.

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莫亚,任郭廷,邓睎远,等.驻景丸加减方对形觉剥夺性近视小鼠视网膜厚度及细胞凋亡的影响.国际眼科杂志, 2021,21(12):2053-2059.

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  • 收稿日期:2021-04-24
  • 最后修改日期:2021-11-10
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  • 在线发布日期: 2021-11-22
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