摘要
目的研究五味子乙素(Sch B)预防环境污染物苯并(a)芘(BaP)致人绒毛膜滋养层细胞HTR8-SVneo氧化损伤的机制。方法以HTR8-SVneo细胞为载体,构建BaP氧化应激模型。实验分为3组,BaP组、Sch B组和空白对照组。MTS法检测细胞存活率,还原酶法检测细胞培养液中一氧化氮(NO)及总一氧化氮合酶(TNOS)含量,用RT-PCR法检测细胞内诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)mRNA水平。结果 BaP组细胞存活率(72.2±0.9)%较对照组显著下降(P<0.05);而0.1、0.5或2μmol/L Sch B+20μmol/L BaP各组的细胞存活率分别为(78.2±1.5)%、(86.5±0.6)%、(93.4±1.0)%,显著高于BaP组(P<0.05)。BaP组细胞培养液中NO、TNOS含量、iNOS、eNOS mRNA表达量和eNOS蛋白表达量均显著高于对照组(P<0.05);而与BaP组相比,Sch B组中NO、TNOS含量,iNOS、eNOS mRNA表达量和eNOS蛋白表达量均随着Sch B剂量的增加逐渐下降(P<0.05)。结论 Sch B可有效预防BaP所致的HTR8-SVneo细胞氧化损伤,其机制可能与NOS/NO系统的平衡有关。
Objective： To explore the mechanism of Schizandrin B preventing oxidative stress injury induced by benzo[a]-pyrene（BaP）in HTRS-SVneo cells. Methods： The model of oxidative stress injury induced by BaP was established with HTR8-SVneo cells. The HTR8-SVneo cells were incubated with various concentrations Sch B for 6 hours. The HTR8- SVneo cells were divided into three groups： BaP group, Seh B group and control group. The cell survival rate was detected by cell proliferation assay（MTS）. NO and TNOS in the cell culture media were detected by reductase method. The mRNA expressions of iNOS and eNOS were determined by RT-PCR. Results： The cell survival rate was significantly declined in BaP group compared with control group（P 〈0.05）. The cell survival rate （78.2 ± 1.5 ） %, （86.5 ±0. 6） %, and （ 93.4 ± 1.0） % respectively in 0.1, 0.5,2 % mol/L Sch B plus 20 tzmol/L BaP were significantly higher than that in BaP group（72.2＋0.9）% （P〈0.05）. The activity of NO and TNOS,and the mRNA levels of iNOS and eNOS were significantly higher than those in control group（P〈0. 05）. The expression of eNOS protein was also significantly increased（P〈0.05）. The activities of NO and TNOS,the mRNA levels of iNOS and eNOS,and eNOSprotein were significantly decreased in Sch B groups along with the concentration of Sch B compared with BaP group（P〈0.05）. Conclusions： Sch B can prevent oxidative stress injury induced by BaP in HTR8-SVneo cells,which may be related NOS/NO system balance.
出处
《生殖医学杂志》
CAS
2015年第7期572-577,共6页
Journal of Reproductive Medicine
基金
国家自然科学基金(81273977)
天津市自然科学基金一般项目(12JCYBJC16200)
