灯盏花素通过调节Bcl-2和Bax表达拮抗高糖诱导的RPE细胞损伤
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黑龙江省博士后资助基金(No.LBH-Z18239)


Scutellarin antagonizing retinal pigment epithelial cell damage induced by high glucose and its molecular mechanism
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Heilongjiang Province Postdoctoral Fundation(No.LBH-Z18239)

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    摘要:

    目的:探讨灯盏花素对高糖环境下人视网膜色素上皮(RPE)细胞氧化应激的影响。

    方法:常规培养RPE细胞,分为对照组(葡萄糖浓度为5.5mmol/L)、高糖组(葡萄糖浓度为30mmol/L)、灯盏花素低浓度组(30mmol/L葡萄糖+1μmol/L灯盏花素)和灯盏花素高浓度组(30mmol/L葡萄糖+10μmol/L灯盏花素)。细胞划痕实验观察RPE细胞的迁移性,CCK-8法检测细胞活性,流式细胞术检测细胞内活性氧(ROS)水平变化,Hoechst染色法观察凋亡细胞比例,Western blot分析各组Bcl-2和Bax的变化。

    结果:细胞划痕实验结果显示,灯盏花素低浓度组和灯盏花素高浓度组RPE细胞形态较高糖组改善,细胞迁移性较高糖组增加; CCK-8结果显示,用1、10μmol/L浓度的灯盏花素处理后,RPE细胞存活率分别提高到61.06%±5.59%和79.81%±7.04%,与高糖组(40.63%±4.72%)比较有差异(P<0.05); 2',7'-二氯二氢荧光素二乙酸酯(H2DCFDA)荧光探针染色结果显示,灯盏花素抑制了RPE细胞内ROS水平; Hoechst染色法显示,1、10μmol/L灯盏花素处理后发生凋亡的RPE细胞数量逐渐减少; Western blot结果显示,灯盏花素增强了Bcl-2蛋白的表达,降低了Bax蛋白的表达。

    结论:灯盏花素可以抑制高糖诱导的RPE细胞氧化损伤和凋亡的发生,为糖尿病视网膜病变的治疗靶点研究提供了理论支持。

    Abstract:

    AIM: To investigate the effect of scutellarin on high glucose-induced human retina pigment epithelium(RPE)cells under high glucose.

    METHODS: RPE cells were cultured and divided into control group(5.5mmol/L glucose), high glucose group(30mmol/L glucose), low concentration scutellarin group(30mmol/L glucose+1μmol/L scutellarin)and high concentration scutellarin group(30mmol/L glucose+10μmol/L scutellarin), cell scratch experiment observed RPE cells migration, cell survival rate were detected by CCK-8 colorimetry, flow cytometry was used to detect the level of ROS, Hoechst staining was used to observe the proportion of apoptotic cells, and Western blot was used to analyze the changes of Bcl-2 and Bax.

    RESULTS: Cell scratch experiment results showed that RPE cells form in low concentration scutellarin group and high concentration scutellarin group were improved than that in high glucose group, cell mobility rate also increased; The CCK-8 results showed that RPE cells survival rate increased to 61.06%±5.59% and 79.81%±7.04% after treated with 1μmol/L and 10μmol/L scutellarin, the difference was statistically significant when compared with high glucose group(40.63%±4.72%, P<0.05); The H2DCFDA fluorescent probe dying showed that scutellarin reduced ROS generation in RPE cells; Hoechst staining showed that the number of apoptosis RPE cells gradually decreased after treatment with 1μmol/L and 10μmol/L scutellarin; Western blot results showed that scutellarin enhanced the expression of Bcl-2 protein and reduced the expression of Bax protein.

    CONCLUSION: Scutellarin could inhibit high glucose-induced oxidative damage and apoptosis in human RPE cells, which provides theoretical support for the research on the therapeutic targets of diabetic retinopathy.

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刘海峰,赵永厚,高源.灯盏花素通过调节Bcl-2和Bax表达拮抗高糖诱导的RPE细胞损伤.国际眼科杂志, 2020,20(2):230-233.

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  • 收稿日期:2019-04-02
  • 最后修改日期:2019-12-30
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  • 在线发布日期: 2020-01-19
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