论文部分内容阅读
目的观察烟酰胺二磷酸腺苷(NADPH)氧化酶的抑制剂能否抑制人晶状体上皮细胞增殖,探讨NADPH氧化酶亚单位NOX的同源异构体在人晶状体上皮细胞mRNA的表达情况。设计实验性研究。研究对象人晶状体上皮细胞永生系(SRA01/04)。方法实验分为4组,SRA01/04中加入表皮生长因子(EGF)(EGF组),加入NADPH氧化酶抑制剂二亚苯基碘(DPI)(DPI组),先后加入DPI和EGF(DPI+EGF组),不加入上述物质为对照组。利用酶标仪和活细胞计数CCK-8试剂盒检测各组人晶状体上皮细胞OD450值。用RT-PCR方法检测NOX的同源异构体在SRA01/04中的表达,将RT-PCR产物测序,利用NCBIBLAST软件对测序结果与cDNA进行序列对比。主要指标人晶状体上皮细胞的OD450值,NOX的同源异构体表达情况及其与人其他组织的序列对比。结果加入CCK-83.5小时后,EGF组比对照组OD450值升高了12.0%(P=0.000);DPI组比对照组OD450值下降了25.5%(P=0.000);DPI+EGF组OD450值比EGF组降低了26.1%(P=0.000)。RT-PCR结果表明,NOX蛋白的5种同源异构体[包括NOX1、NOX2(gp91phox)、NOX3、NOX4、NOX5]的mRNA在SRA01/04中均有表达。人晶状体上皮细胞中的NOX1-5的序列与人其他组织的相似性分别为100%、100%、99.7%、100%、100%。结论NADPH氧化酶可能促进人晶状体上皮细胞的增殖;NOX同源异构体NOX1、NOX2、NOX3、NOX4、NOX5的mRNA在人晶状体上皮细胞SRA01/04中均有表达,其中NOX1明显弱于其他四种NOX。
Objective To observe whether inhibitors of nicotinamide adenosine diphosphate (NADPH) oxidase can inhibit the proliferation of human lens epithelial cells and investigate the mRNA expression of NOX isoforms of NADPH oxidase subunits in human lens epithelial cells. Design experimental research. Human lens epithelial cells immortalized (SRA01 / 04). Methods The experiment was divided into 4 groups: epidermal growth factor (EGF) was added to SRA01 / 04 and DPI (DPI group), an inhibitor of NADPH oxidase, DPI and EGF EGF group), without adding the above substance as a control group. The OD450 values of human lens epithelial cells in each group were detected by using microplate reader and viable cell count CCK-8 kit. The expression of NOX isoforms in SRA01 / 04 was detected by RT-PCR. The RT-PCR products were sequenced. The sequencing results were compared with that of cDNA by NCBIBLAST software. The main index of human lens epithelial cells OD450 value, NOX homeobox expression and sequence comparison with other human tissues. Results OD450 increased by 12.0% (P = 0.000) in EGF group compared with control group after CCK-83.5 hours. The OD450 value decreased by 25.5% (P = 0.000) in DPI group compared with control group. OD450 value in DPI + EGF group The EGF group decreased by 26.1% (P = 0.000). The results of RT-PCR showed that the mRNAs of NOX1, NOX1, NOX2 (gp91phox), NOX3, NOX4 and NOX5 were all expressed in SRA01 / 04. The sequence of NOX1-5 in human lens epithelial cells was 100%, 100%, 99.7%, 100%, 100% similar to other human tissues, respectively. Conclusion NADPH oxidase may promote the proliferation of human lens epithelial cells. The NOX1, NOX2, NOX3, NOX4 and NOX5 mRNA expressions in human lens epithelial cells SRA01 / 04 were significantly higher than those in NOX1 NOX.