α-倒捻子素对过氧化氢诱导的人视网膜色素上皮细胞损伤的保护作用
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国家重点基础研究发展计划(973计划 No.2013CB967500; No.2011CB510200); 国家自然科学基金项目面上项目(No.81170855); 江苏省临床医学科技专项--重点病种的规范化诊疗研究(No.BL2014089); “六大人才高峰”D类资助项目(No.2013-WSW-015)


Protective effects of α-Mangostin on injury of human retinal pigment epithelium cells induced by hydrogen peroxide
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National Basic Research Program of China(973 Program,No.2013CB967500 and No.2011CB510200); General Project of National Natural Science Foundation of China(No.81170855); Jiangsu Provincial Special Program of Medical Science(No. BL2014089); Six Talent Peaks Project in Jiangsu Province(No. 2013-WSW-015)

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    摘要:

    目的:研究α-倒捻子素对过氧化氢(H2O2)诱导的人视网膜色素上皮细胞株ARPE-19氧化损伤的保护作用。

    方法:分别用不同浓度的α-倒捻子素和H2O2处理ARPE-19,CCK8法检测α-倒捻子素和H2O2对ARPE-19细胞毒性作用。不同浓度的α-倒捻子素预处理ARPE-19,再用200μmol/L H2O2处理24h,观察细胞活性变化。流式细胞仪检测细胞活性氧(reactive oxygen species, ROS)水平变化,免疫印迹法(Western blot)检测NF-κB蛋白表达变化。

    结果:CCK8法检测结果显示:当α-倒捻子素浓度在0~12μmol/L时,ARPE-19活性无明显变化; 当浓度达到16μmol/L时,细胞活性开始下降(P<0.05)。H2O2诱导后,当α-倒捻子素浓度在0~16μmol/L之内时,α-倒捻子素预处理均可提高ARPE-19细胞活性。ROS结果表示:H2O2诱导后,ROS表达量增高(P<0.05); 8和12μmol/L α-倒捻子素预处理,均可有效清除H2O2诱导产生的ROS(P<0.05)。Western blot结果显示:H2O2诱导后NF-κB蛋白表达增高(P<0.05),12μmol/Lα-倒捻子素预处理可以继续上调H2O2诱导后ARPE-19的NF-κB蛋白表达(P<0.05)。

    结论:H2O2诱导ARPE-19细胞氧化损伤,造成细胞活性下降,ROS表达量增加,经过α-倒捻子素预处理后,可有效提高细胞活性,清除ROS,活化NF-κB。

    Abstract:

    AIM: To study protective effects of α-Mangostin in human retinal pigment epithelial(RPE)cells induced by hydrogen peroxide(H2O2).

    METHODS:ARPE-19 cells were treated with different concentrations of α-Mangostin and H2O2.The effect of α-Mangostin and H2O2 respectively on cell activity was detected by CCK8. ARPE-19 cells were pretreated with different concentrations of α-Mangostin for 24h before they were administrated with 200μmol/L H2O2for another 24h.Then the changes of cell activity were observed. The expression of reactive oxygen species(ROS)level was detected by flow cytometry(FCM)and the expression of NF-κB protein was measured by Western blot analysis.

    RESULTS:CCK8 examination results showed that: within 0~12μmol/L, α-Mangostin had no damage effects on cell activity. When the concentration of 16μmol/L, cell viability began to decrease(P<0.05).And α-Mangostin pretreatment gradually increased cell viability of ARPE-19 induced by H2O2 when the concentrations of α-Mangostin were within 0~16μmol/L. ROS results showed: the expression of ROS level significantly increased after H2O2 induced(P<0.05); 8 and 12μmol/L α-Mangostin pretreatment down-regulated ROS expression of ARPE-19 induced by H2O2(P<0.05). Western blot results showed that the expression of NF-κB protein after H2O2 induced increased(P<0.05); 12μmol/L α-Mangostin pretreatment up-regulated NF-κB of ARPE-19 induced by H2O2(P<0.05).

    CONCLUSION: H2O2 induced oxidative damage in RPE cells by decreasing cell viability and increasing the expression of ROS level. α-Mangostin can protect RPE cells from the injury of H2O2, the mechanism may be related to the clear of ROS and the activation of NF-κB.

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苏途,方圆,谢平,等.α-倒捻子素对过氧化氢诱导的人视网膜色素上皮细胞损伤的保护作用.国际眼科杂志, 2015,15(6):959-962.

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  • 收稿日期:2015-03-16
  • 最后修改日期:2015-05-15
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  • 在线发布日期: 2015-06-01
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