A Novel Chemiluminescence System for Detecting and Analyzing Oxidative Damaged RNA by Hydroxyl Radical and Its Molecular Mechanism

CHEN Ji-wu; ZHU Zhen-qin; WANG Xi-lu; ZHAN Xuan-zhi; YANG Xiao-ning

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A Novel Chemiluminescence System for Detecting and Analyzing Oxidative Damaged RNA by Hydroxyl Radical and Its Molecular Mechanism

更新时间：2020-08-11

- A Novel Chemiluminescence System for Detecting and Analyzing Oxidative Damaged RNA by Hydroxyl Radical and Its Molecular Mechanism
- Chinese Journal of Luminescence Vol. 23, Issue 1, Pages: 71-74(2002)
- 作者机构：
  
  华东师范大学生命科学学院, 上海, 200062
- 作者简介：
- 基金信息：
- DOI：
  CLC： Q527.4;O657.3
- Received：14 May 2001，
  
  Revised：2001-7-15，
  
  Published：20 January 2002
- 稿件说明：
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陈季武, 朱振勤, 王喜露, 詹选治, 杨晓宁. 一种新颖的测定和分析羟自由基氧化损伤RNA及其分子机理的化学发光体系[J]. 发光学报, 2002,23(1): 71-74

CHEN Ji-wu, ZHU Zhen-qin, WANG Xi-lu, ZHAN Xuan-zhi, YANG Xiao-ning. A Novel Chemiluminescence System for Detecting and Analyzing Oxidative Damaged RNA by Hydroxyl Radical and Its Molecular Mechanism[J]. Chinese Journal of Luminescence, 2002,23(1): 71-74
陈季武, 朱振勤, 王喜露, 詹选治, 杨晓宁. 一种新颖的测定和分析羟自由基氧化损伤RNA及其分子机理的化学发光体系[J]. 发光学报, 2002,23(1): 71-74 DOI：

CHEN Ji-wu, ZHU Zhen-qin, WANG Xi-lu, ZHAN Xuan-zhi, YANG Xiao-ning. A Novel Chemiluminescence System for Detecting and Analyzing Oxidative Damaged RNA by Hydroxyl Radical and Its Molecular Mechanism[J]. Chinese Journal of Luminescence, 2002,23(1): 71-74 DOI：

摘要

近年来

羟自由基(

OH)对DNA氧化损伤已受到广泛关注.但是很少研究

OH对RNA的氧化损伤.其实

RNA与DNA一样

也是核酸的两大组分之一

也有许多重要功能.所以

OH攻击RNA也会引起严重后果

会造成细胞功能衰退甚至细胞死亡等.为此

我们建立了Vit.C-CuSO

-Phen-H

-RNA这一产生和测定

OH氧化损伤RNA的化学发光体系

以便加强

OH氧化损伤RNA的研究.通过对本体系测定条件的研究

得出了本体系最佳组方是:Vit.C、CuSO

、Phen、H

和RNA

浓度分别为350μmol/L、55μmol/L、350μmol/L、0.2mol/L和20μg/mL

体系pH为5.5

体系终体积为1mL.随后

利用本体系检测了槲皮素、咖啡酸、黄芩甙和芦丁抗

OH氧化损伤RNA的作用

发现这四种抗氧化剂均能有效抑制

OH对RNA的氧化损伤

从而也证明了本体系的可行性和实用性.我们也利用此体系和特异抗氧化剂分析了

OH氧化损伤RNA的分子机理

结果发现

OH清除剂硫脲几乎抑制全部发光

推测是因硫脲清除了引发剂

OH而不能引发RNA氧化损伤所致;过氧化氢酶几乎能抑制全部发光

推测是因过氧化氢酶清除了H

而不能产生引发剂

OH所致;O

清除剂SOD只能抑制小部分发光;

清除剂叠氮化钠和苯甲酸都能抑制绝大部分发光.这些事实提示

OH是RNA氧化损伤的引发剂;O

只是导致RNA氧化损伤的次要因素

才是导致RNA氧化损伤的最主要因素.

Abstract

In recent years

oxidative damaged DNA by hydroxyl radical(

OH)has been received much attention.But the reports on oxidative damaged RNA by

OH were quite limited.In fact

RNA

being the same as DNA

is also one of big two components for nucleic acid.It also has many important functions.Therefore

RNA attacked by

OH leads to serious consequence

which bring about decline of the cell activity even death of the cell.Thus a chemiluminescence system for generating and detecting oxidative damaged RNA by

OH was made

which initiated with Vit.C/CuSO

/Phen/H

/RNA

so as to strengthen the study on oxidative damaged RNA by

OH.Through the investigation on detecting conditions of the system

optimal scheme of the system was obtained:concentration of Vit.C

CuSO

Phen

and RNA were 350μmol/L

55μmol/L

350μmol/L

0.2mol/L and 20μg/mL

respectively.The pH of the system was 5.5.The final volume was 1mL.Using the system

we detected the effects that quercetin

caffeic acid

baicalin and rutin resisted oxidative damaged RNA by

OH and found that the four antioxidants could effectively inhibited oxidative damaged RNA by

OH.These facts also proved feasibility and practicality of the system.Molecular mechanism for oxidative damaged RNA by

OH was analyzed by the system and special antioxidants.The results indicated that sulfocarbamide which was a

OH scavenger nearly inhibited whole chemiluminescence.The reason for the phenomenon was that RNA oxidative damage was not initiated only because sulfocarbamide scavenged

an initiator;catalase also inhibited whole chemiluminescence

revealing that

OH couldn't be generated due to H

removed by catalase.SOD

a O

scavenger

could only inhibit a small part of chemiluminescence

whereas NaN

or benzene carboxylic acid

scavenger

could inhibit most of chemiluminescence.These results suggested that

OH is an initator;but O

is a minor factor

is a major factor for the RNA oxidative damage.

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