阿柏西普对体外培养的视网膜Müller细胞膜离子通道的影响
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2015~2016年度湖北省卫生计生西医类一般资助项目(No.WJ2015MB218)


Effects of Aflibercept on ion channel of retinal Müller cell membrane cultured in vitro
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Research Project of Hubei Provincial Health and Family Planning Commission in 2015-2016(No.WJ2015MB218)

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    摘要:

    目的:探讨阿柏西普对体外培养的高糖环境下视网膜Müller细胞膜K+通道的影响。

    方法:人Müller细胞分为3组:对照组(常规DMEM培养基培养)、高糖组(高糖DMEM培养基培养)、实验组(高糖DMEM培养基和100μmol/L 阿柏西普处理)。采用荧光探针检测细胞K+浓度,MTT法检测细胞存活率,流式细胞仪检测细胞凋亡率,Western blot法检测Müller细胞caspase-3蛋白水平。

    结果:Müller细胞培养48h后呈现谷氨酰胺合成酶(GS)阳性,纯化度在90%以上。荧光检测显示对照组、高糖组、实验组K+相对浓度分别为(2.14±0.44)%、(23.11±4.39)%、(5.20±0.92)%,细胞存活率分别为(100.00±0.00)%、(73.24±4.13)%、(85.22±5.33)%,细胞凋亡率分别为(5.03±1.91)%、(26.73±3.14)%、(16.63±2.73)%(均P<0.05)。 与对照组相比,高糖组Müller细胞内caspase-3蛋白水平显著上升(P<0.05); 与高糖组Müller细胞相比,实验组Müller细胞内caspase-3蛋白水平显著降低(P<0.05)。

    结论:阿柏西普可抑制体外培养的高糖环境下视网膜Müller细胞膜K+通道,抑制高糖诱导的Müller细胞凋亡,降低caspase-3蛋白表达水平,促进细胞增殖。

    Abstract:

    AIM: To investigate the effects of Aflibercept on the K+ channel of retinal Müller cell membrane cultured in vitro.

    METHODS: Human Müller cells were divided into 3 groups(control group, high glucose group and experimental group). The control group were cultured in conventional DMEM medium; the high glucose group were cultured in high glucose DMEM medium; the experimental group were cultured with high glucose DMEM medium and 100μmol/L Aflibercept, and the K+ concentration of the cells were detected by MQAE, and the cell survival were detected by MTT assay, the flow cytometry were used to detect apoptosis rates, Western blot analysis were used to detect the Müller cell caspase-3 protein levels.

    RESULTS: The Müller cells were positive for glutamine synthetase(GS)after 48h of culture, and the purification degree were above 90%. The relative concentrations of K+ in the control group, high glucose group and experimental group were(2.14±0.44)%,(23.11±4.39)%,(5.20±0.92)%, and cell viability were(100.00±0.00)%, respectively(73.24±4.13)%,(85.22±5.33)%, the apoptosis rates were(5.03±1.91)%,(26.73±3.14)%,(16.63±2.73)%, and compared the differences between two groups were statistically significant(P<0.05). Compared with the control group, the level of caspase-3 protein in the high glucose group Müller cells were increased significantly(P<0.05); compared with the high glucose group Müller cells, the caspase-3 protein level in the experimental group Müller cells were decreased significantly(P<0.05).

    CONCLUSION: Aflibercept can inhibit the K+ channel of retinal Müller cells in vitro, inhibit the apoptosis of Müller cells induced by high glucose, decrease the expression of caspase-3 protein, and promote cell proliferation.

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雷琦峰,蔡维.阿柏西普对体外培养的视网膜Müller细胞膜离子通道的影响.国际眼科杂志, 2019,19(4):547-550.

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  • 收稿日期:2018-11-08
  • 最后修改日期:2019-03-12
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  • 在线发布日期: 2019-03-22
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