摘要
目的:探讨血红素氧合酶(HO)两种同工酶HO-1和HO-2对肿瘤坏死因子-α(TNF-α)诱导胎盘组织损伤的保护作用。方法:用胎盘组织块培养(n=6)方法,以乳酸脱氢酶(LDH)释放至培养基质中的量作为毒性指数,用免疫印迹法对HO的诱导表达进行半定量分析。分7组,分别加入10μl DMEM液(对照组);底物5μmol/L血红素(Hemin组);TNF-α,终浓度为50ng/ml(TNF-α组);血红素加TNF-α(H+T组);加入HO的特异性抑制剂SnPP 20μmol/L(SnPP组);SnPP+TNF-α(S+T组);SnPP+血红素+TNF-α(S+H+T组)。各组组织块均培养12h后进行分析。结果:(1)TNFα组细胞毒指数明显增加(0.78±0.23),与对照组(0.44±0.12)相比有显著差异(P<0.01);H+T组细胞毒指数(0.53±0.13)比TNF-α组明显降低(P<0.05)。单独加入底物血红素和加入SnPP对细胞毒指数无明显影响(P>0.05)。T+S组和T+S+H组的细胞毒指数与TNF-α组无显著差异(P>0.05);(2)免疫印迹分析显示,TNF-α组导致HO-1表达下调(0.37±0.21),与对照组(0.83±0.27)的差异有显著性(P<0.05);而H+T组HO-1表达上调(0.92±0.34),与TNF-α组的差异有显著性(P<0.05);HO-2的表达无明显变化。结论:血红素诱导的HO-1表达能对抗TNF-α导致的胎盘组织损伤,HO抑制剂SnPP可阻断其作用。血红素诱导对HO-2表达无明显影响。
Objective：To investigate the effect of HO activation on tumor necrosis factor-alpha（TNF-α） induced placental damage. Method：Using placental villous explant experiment, cytotoxicity could be measured as leakage of lactate dehydrogenase （LDH） into the culture medium from tissues. Western blots was resorted to demonstrate the effect of HO activation on tumor necrosis factor-alpha （TNF-α） induced placental damage. There were 7groups, （1） control group ; （2） hemin ; （3） TNF α ; （ 4 ） hemin ＋ TNF α; （ 5 ） SnPP; （ 6 ） SnPP ＋ TNF-α ; （7） SnPP ＋ heroin ＋ TNF-α. Result： （ 1 ） Placental villous explanted incubated with TNF-α caused cytotoxicity index（0.78 ± 0.23 ） significantly increased （ P 〈 0.01 ）. The cytotoxicity index（0.53 ± 0.13 ） was significantly reduced when preincubated with heroin for 2h prior to the addition of TNFα （P 〈 0.05）. Incubated with heroin or SnPP overnight did not increased LDH leakage significantly （ P 〉 0. 05 ）. In the presence of SnPP, the protective effect of hemin on TNF-α induced damage was totally abolished, the cytotoxicity index had no significantly difference than those in TNF-α group （ P 〉 0.05 ）. （ 2 ） Western blots analysis showed that, in the presence of TNF-α, heroin （ H ＋ T） caused an up-regulation of HO-1 expression （0.92 ±0. 34）. In contrast,TNFα itself （0.37 ± 0.21 ） decreased HO-1 expression（ P 〈 0.05 ）. Compared with control, the expression of HO-2 had no difference. Conclusion： Induction of HO-1 cause a significant attenuation of TNF-α-mediated cellular damage in placental villous explants,and HO inhibitor SnPP reverses the effect. Expression of HO-2 has no difference.
出处
《现代妇产科进展》
CSCD
北大核心
2008年第12期916-919,共4页
Progress in Obstetrics and Gynecology
关键词
妊娠并发症
心血管
肿瘤坏死因子Α
胎盘
血红素氧化酶
Pregnancy complication, cardiovascular
Tumor necrosis factor alpha
Placenta
Heme oxygenase
作者简介
第一作者：程国梅（1969-），女，郑州大学第三附属医院妇产科副主任医师、博士。主要研究方向：围产医学。
