Induction of renal cell line apoptosis by hydrogen peroxide

Khalil Alhalfawy, Bahgat A. Elfiky, A. Zahran, M. Zayed
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Abstract

Background: Hydrogen peroxide is currently the most widely used as an apoptosis inducer due to its broad cytotoxic efficacy against nearly all cell types. cytotoxicity is achieved over a wide range of doses Objective: To evaluate the cytotoxic effect of hydrogen peroxide on renal cell lines by detecting RIPK1. Methods: In this study, we used a Vero cell line treated with H 2 O 2 at concentrations 0.1 mM, 0.2 mM, 0.4 mM, 0.8 mM, and 1.6 mM and examined after 30 min,1hour,2hours,3hours, 4hours and 5 hours. by using MTT assay to detect cytotoxicity to cell line (by detecting cell viability). Spectrophotometrically measure the absorbance at a wavelength of 570 nm. Measure the background absorbance of multi-well plates at 690 nm and subtract from the 450 nm measurement. Sub-lethal dose to renal cell line is one treated with 1.6 mM for 5 hours. groups group 1: renal cell line as control which not treated by H2O2. group 2: Sub-lethal which renal cell line treated by concentration 1.6 mM of H2O2 for 5 hours. Results: H2O2 is cytotoxic to renal cell line by concentration from 0.1 mM to 1.6 mM. RIPK1 gene expressed in renal cell line treated by H2O2. The sublethal dose reached 1.6 mM for 5 hours. There is a significant difference between the 2 groups by detecting the expression of the RIPK1 gene.
过氧化氢诱导肾细胞系凋亡
背景:过氧化氢对几乎所有类型的细胞都具有广泛的细胞毒性,是目前应用最广泛的细胞凋亡诱导剂。目的:通过检测RIPK1,评价过氧化氢对肾细胞系的细胞毒性作用。方法:采用浓度分别为0.1 mM、0.2 mM、0.4 mM、0.8 mM、1.6 mM的h2o2处理Vero细胞株,分别于30min、1h、2h、3h、4h、5h后进行检测。采用MTT法检测细胞系的细胞毒性(通过检测细胞活力)。分光光度法测量波长为570nm处的吸光度。在690 nm处测量多孔板的背景吸光度,并从450 nm测量值中减去。对肾细胞系的亚致死剂量为1.6 mM,作用5小时。组1:未经H2O2处理的肾细胞系作为对照。2组:浓度为1.6 mM的H2O2处理肾细胞株5小时后亚致死。结果:H2O2浓度在0.1 mM ~ 1.6 mM范围内对肾细胞株具有细胞毒性。亚致死剂量达1.6 mM,持续5小时。通过检测RIPK1基因的表达,两组间存在显著差异。
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