CAJ | 학술논문

目的：研究经香烟烟雾提取物（CSE）刺激后大鼠膈肌细胞8-羟基脱氧鸟苷（8-OHdG）含量的变化以及细胞内活性氧（ROS）水平和8-羟基鸟嘌呤DNA 糖苷酶1（OGG1）表达对8-OHdG含量变化的影响。方法用不同浓度CSE分别刺激大鼠膈肌细胞24、48和72h后，采用高效液相色谱-电化学法（HPLC-ECD）检测大鼠膈肌细胞8-OHdG的含量，流式细胞术检测大鼠膈肌细胞ROS 水平，采用实时定量PCR检测OGG1 mRNA水平，用Western印迹法检测OGG1蛋白水平。结果经相同CSE浓度分别刺激大鼠膈肌细胞24、48和72h后,大鼠膈肌细胞中8-OHdG含量、ROS水平及OGG1 mRNA和蛋白水平均无明显差异。但经不同浓度CSE刺激相同时间后，10.00% CSE和20.00% CSE刺激组大鼠膈肌细胞8-OHdG含量明显高于对照组和5.00% CSE刺激组（P＜0.05）；大鼠膈肌细胞内ROS水平、OGG1 mRNA和蛋白水平较对照组均明显升高（P＜0.05）；大鼠膈肌细胞8-OHdG含量与其ROS水平呈明显正相关（r＝0.826，P＝0.000）；经CSE刺激后，大鼠膈肌细胞OGG1 mRNA和蛋白水平均高于对照组（P＜0.05），但大鼠膈肌细胞8-OHdG含量与其OGG1 mRNA和蛋白水平无明显相关（r＝0.373，P＝0.254；r＝0.329，P＝0.296）。结论 CSE刺激可引起大鼠膈肌细胞8-OHdG升高，8-OHdG含量与ROS水平有关，但与其修复酶OGG1表达水平无明显相关。
목적：연구경향연연무제취물（CSE）자격후대서격기세포8-간기탈양조감（8-OHdG）함량적변화이급세포내활성양（ROS）수평화8-간기조표령DNA 당감매1（OGG1）표체대8-OHdG함량변화적영향。방법용불동농도CSE분별자격대서격기세포24、48화72h후，채용고효액상색보-전화학법（HPLC-ECD）검측대서격기세포8-OHdG적함량，류식세포술검측대서격기세포ROS 수평，채용실시정량PCR검측OGG1 mRNA수평，용Western인적법검측OGG1단백수평。결과경상동CSE농도분별자격대서격기세포24、48화72h후,대서격기세포중8-OHdG함량、ROS수평급OGG1 mRNA화단백수평균무명현차이。단경불동농도CSE자격상동시간후，10.00% CSE화20.00% CSE자격조대서격기세포8-OHdG함량명현고우대조조화5.00% CSE자격조（P＜0.05）；대서격기세포내ROS수평、OGG1 mRNA화단백수평교대조조균명현승고（P＜0.05）；대서격기세포8-OHdG함량여기ROS수평정명현정상관（r＝0.826，P＝0.000）；경CSE자격후，대서격기세포OGG1 mRNA화단백수평균고우대조조（P＜0.05），단대서격기세포8-OHdG함량여기OGG1 mRNA화단백수평무명현상관（r＝0.373，P＝0.254；r＝0.329，P＝0.296）。결론 CSE자격가인기대서격기세포8-OHdG승고，8-OHdG함량여ROS수평유관，단여기수복매OGG1표체수평무명현상관。
Objective To study the effect of cigarette smoke extract(CSE) on the diaphragm cells of rats in 8-hydroxydeoxyguanosine(8-OHdG) content and the relationship between 8-OHdG content and level of reactive oxygen species(ROS) and 8-oxoguanine DNA glycosylase-1(OGG1). Methods The diaphragm cells of rats were cultured and stimulated by CSE for different time periods and at different concentrations. 8-OHdG was detected by high performance liquid chromatography with electrochemical detection(HPLC-ECD). The level of ROS was determined by fluorescent probe of 2’, 7’-dichlorodihydrofluorescein diacetate (DCFH-DA). The expression of OGG1 mRNA was tested by real-time PCR. The expression of OGG1 protein was tested by Western blot. Results There was no significant difference in the 8-OHdG, ROS, OGG1 mRNA and protein levels after CSE treatment for different time periods. However, the 8-OHdG content of the diaphragm cells induced by 10.00% CSE and 20.00% CSE was significantly increased compared with the diaphragm cells induced by 5.00% CSE and control groups(P<0.05). There was no significant difference in 8-OHdG content between 5.00% CSE and control groups(P>0.05). Compared with control, the intracellular ROS level was dramatically increased in CSE-treated groups(P<0.05), and the 8-OHdG content was increased according to the level of ROS(r=0.826, P=0.000). Compared with control, the expression of OGG1 mRNA and protein were dramatically increased in CSE-treated groups(P<0.05), however, the 8-OHdG content has but not with the expression of OGG1 mRNA and protein(r=0.373, P=0.254;r=0.329, P=0.296). Conclusions CSE could increase the level of 8-OHdG in the diaphragm cells of rats. The 8-OHdG content has correlation with ROS level, but not with the expression of OGG1.