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通过下调HtrA1抑制蓝光照射下人视网膜色素上皮细胞的增殖、迁移和凋亡。

Inhibition of cell proliferation, migration and apoptosis in blue-light illuminated human retinal pigment epithelium cells by down-regulation of HtrA1.

作者信息

Yu Tian, Chen Chang-Zheng, Xing Yi-Qiao

机构信息

Eye Center, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.

出版信息

Int J Ophthalmol. 2017 Apr 18;10(4):524-529. doi: 10.18240/ijo.2017.04.04. eCollection 2017.

Abstract

AIM

To investigate the effect of HtrA1 on the proliferation, migration and apoptosis of human retinal pigment epithelium (RPE) cells in the light injured model, as well as the expression of the apoptosis related molecules.

METHODS

The human RPE cell line ARPE-19 was exposed to blue light to establish the light injured model. The cells were transfected with HtrA1 siRNA to knockdown HtrA1 expression. Subsequent expression of HtrA1 was determined by real-time polymerase chain reaction (RT-PCR) and Western blot, respectively. Changes in cell proliferation, migration and apoptosis were assessed by cell counting kit-8 (CCK-8), Transwell assay and flow cytometry respectively, as well as changes in the mRNA and protein levels of Bax, Caspase-3 and Bcl-2 expression.

RESULTS

HtrA1 was highly expressed in ARPE-19 cells after blue light irradiation. Knockdown of HtrA1 expression inhibited the proliferation, migration and apoptosis of the blue-light-irradiated ARPE-19 cells (<0.05). Bax and Caspase-3 expression were significantly reduced both at mRNA and protein levels (<0.05) after siRNA treatment. Bcl-2 expression significantly increased in blue-light-irradiated ARPE-19 cells after siRNA interference (<0.05).

CONCLUSION

Silence of HtrA1 may inhibit the proliferation, migration and apoptosis of ARPE-19 cells in light injured model. Moreover, HtrA1 suppression in blue-light-irradiated ARPE-19 cells may ameliorate cell apoptosis through down-regulation of Bax and Caspase-3, and up-regulation of Bcl-2 expression.

摘要

目的

研究HtrA1对光损伤模型中人类视网膜色素上皮(RPE)细胞增殖、迁移和凋亡的影响,以及凋亡相关分子的表达。

方法

将人类RPE细胞系ARPE-19暴露于蓝光下以建立光损伤模型。用HtrA1 siRNA转染细胞以敲低HtrA1表达。随后分别通过实时聚合酶链反应(RT-PCR)和蛋白质免疫印迹法测定HtrA1的表达。分别通过细胞计数试剂盒-8(CCK-8)、Transwell实验和流式细胞术评估细胞增殖、迁移和凋亡的变化,以及Bax、Caspase-3和Bcl-2表达的mRNA和蛋白质水平的变化。

结果

蓝光照射后,HtrA1在ARPE-19细胞中高表达。敲低HtrA1表达可抑制蓝光照射的ARPE-19细胞的增殖、迁移和凋亡(<0.05)。siRNA处理后,Bax和Caspase-3的表达在mRNA和蛋白质水平均显著降低(<0.05)。siRNA干扰后,蓝光照射的ARPE-19细胞中Bcl-2表达显著增加(<0.05)。

结论

沉默HtrA1可能抑制光损伤模型中ARPE-19细胞的增殖、迁移和凋亡。此外,抑制蓝光照射的ARPE-19细胞中的HtrA1可能通过下调Bax和Caspase-3以及上调Bcl-2表达来改善细胞凋亡。

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