MsrB1对ONOO~-诱导的人晶状体上皮细胞周期的影响

发布时间：2017-12-27 07:46

本文关键词：MsrB1对ONOO~-诱导的人晶状体上皮细胞周期的影响　出处：《眼科新进展》2015年11期 　论文类型：期刊论文

【摘要】：目的探讨蛋氨酸亚砜还原酶B1(methionine sulfoxide reductase B1,MsrB1)基因沉默对过氧亚硝酸根(ONOO-)诱导的人晶状体上皮细胞(human lens epithelial cells,HLEC)细胞周期的影响和ERK信号通路在其中的调节作用。方法将培养的HLEC分为两组:正常组和MsrB1基因沉默组,分别用0μmol·L-1、100μmol·L-1、200μmol·L-1和300μmol·L-1 ONOO-处理20 min后,继续培养12 h。用RT-PCR法检测MsrB1基因表达水平变化,流式细胞仪检测细胞周期水平变化,Western blot法检测MsrB1基因沉默和ONOO-对pERK表达水平的影响。结果 300μmol·L-1 ONOO-处理HLEC会导致MsrB1表达水平显著下降(P0.01),当MsrB1基因沉默细胞经300μmol·L-1ONOO-处理后,MsrB1表达水平进一步显著下调(P0.05)。经200μmol·L-1或300μmol·L-1 ONOO-处理,HLEC细胞周期分别阻滞在G2/M期和S期,当MsrB1基因沉默细胞经200μmol·L-1或300μmol·L-1 ONOO-处理后,更多的细胞阻滞在G2/M期或S期(P0.05)。pERK检测结果表明,MsrB1基因沉默前后经ONOO-处理导致的细胞周期阻滞和pERK表达水平显著下降有关(P0.01)。结论 ONOO-可以下调HLEC MsrB1表达水平,MsrB1基因沉默细胞经ONOO-处理,pERK表达水平显著下降,导致更多细胞阻滞在G2/M期和S期。
[Abstract]:Objective to investigate the methionine sulfoxide reductase B1 (methionine sulfoxide reductase B1, MsrB1) gene silencing on peroxynitrite (ONOO-) in human lens epithelial cells induced by (human lens epithelial cells, HLEC) on cell cycle and ERK signaling pathway in the regulation of the. Methods the cultured HLEC were divided into two groups: normal group and MsrB1 gene silencing group. They were treated with 0 mol mol L-1, 100 L-1 mol L-1, 200 mol mol L-1 and 300 mol mol L-1 ONOO- 20, respectively. The expression level of MsrB1 gene was detected by RT-PCR method. The cell cycle level was detected by flow cytometry. The effects of MsrB1 gene silencing and ONOO- on pERK expression level were detected by Western blot. Results the expression level of MsrB1 was significantly decreased when treated with 300 HLEC mol L-1 ONOO- (P0.01). When MsrB1 gene silenced cells were treated with 300 mol mol L-1ONOO-, the expression level of MsrB1 was further down regulated (P0.05). After 200, mol, L-1 or 300 mol mol L-1 ONOO- treatment, HLEC cell cycle was blocked at G2/M stage and S stage respectively. When MsrB1 gene silenced cells were treated with 200 mol mol or HLEC or HLEC, more cells were arrested in the "phase" or "phase". The results of pERK detection showed that the cell cycle arrest caused by ONOO- treatment before and after the MsrB1 gene silencing was significantly related to the decrease of pERK expression level (P0.01). Conclusion ONOO- can down regulate the expression level of HLEC MsrB1. MsrB1 gene silencing cells after ONOO- treatment, pERK expression level decreased significantly, resulting in more cell arrest in G2/M and S phase.
【作者单位】：贵州医科大学生物与工程学院;
【基金】：贵州省科学技术基金资助(编号:黔科合J-2014-2028) 贵阳医学院博士启动基金(编号:YJ2014-8)~~
【分类号】：R776.1
【正文快照】： 白内障已经成为大多数国家致盲和视力残疾的主要因素,而且也是我国老年人群致盲的首要原因[1-4]。研究表明,过氧亚硝酸根(ONOO-)导致的氧化和硝化应激介导了白内障的病理过程[5-8]。以前的研究发现,蛋氨酸亚砜还原酶B1(methioninesulfoxide reductase B1,MsrB1)在抵抗人晶状体

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