【作者】 邢志伟；

【导师】 赵富玺；

【作者基本信息】 山西医科大学 ， 免疫学， 2009， 硕士

【摘要】 目的:早产是妊娠常见的并发症之一,感染是引起早产的最常见原因。本实验选择了脂多糖(Lipopolysaccharide, LPS)致感染性早产这一常见的妊娠并发症作为研究对象,通过研究感染性早产小鼠胎盘Toll样受体(4Toll-Like Receptor 4,TLR4),核转录因子-kappaB p65(nuclear factor-kappaB p65,NF-κBp65)和肿瘤坏死因子-α(Tumor Necrosis Factor-alpha,TNF-α)的表达以及N-乙酰半胱氨酸(N-acetylcysteine,NAC)的干预影响,探讨一种预防小鼠感染性早产发生的新策略,为临床上感染性早产的早期预防及早期治疗提供理论依据。方法:(1)孕鼠随机分为4组:对照组(NS)、模型组(LPS)、预防组(NAC+LPS)和治疗组(LPS+NAC);(2)复制小鼠的感染性早产模型,观察各组孕鼠发生早产的时间,记录早产共分为5个时间段(0-12h,12-24h,24-36h,36-48h,48h-);(3)利用逆转录-聚合酶链反应(RT-PCR)方法检测早产小鼠胎盘TLR4,NF-κBp65,TNF-αmRNA的表达,分析三者与感染性早产的关系,以及NAC干预后TLR4,NF-κBp65和TNF-αmRNA的表达变化;(4)利用免疫组化方法检测早产小鼠胎盘TLR4,NF-κBp65,TNF-α蛋白的表达,以及NAC干预后TLR4,NF-κBp65和TNF-α蛋白的表达情况。结果:(1)对照组无早产的发生, LPS组和对照组相比P=0.001,表明感染性早产模型成功建立,预防组与LPS模型组早产率比较P=0.041,早产率明显下降;(2)对照组都有TLR4、NF-κBp65、TNF-α的少量表达,LPS模型组和对照组相比三者表达都显著升高(P<0.05) ;(3)预防组和模型组相比,TLR4表达无明显变化,而NF-κBp65和TNF-α表达明显减少(P<0.05),并且早产率也明显下降(P<0.05);(4)治疗组和模型组相比无统计学意义(P>0.05)。结论:感染性早产模型成功建立,NAC可以降低感染性早产小鼠胎盘NF-κBp65和TNF-α的表达,在感染性早产的预防中有一定作用。更多还原

【Abstract】 Objective: Preterm birth is a common complication of pregnancy and infections are caused by the most common cause of preterm birth. We select LPS-induced preterm birth with infection as a research object in this experiment. By studying Toll-Like Receptor 4(TLR4), nuclear factor -kappaBp65(NF-κBp65), Tumor Necrosis Factor-alpha(TNF-α)expression in the mouse placenta of preterm birth, as well as the impact of N-Acetylcysteine(NAC) intervention, new preventions of premature birth occur in the future are explored. In addition, we may provide a theoretical basis for early clinical prevention and treatment of preterm birth with infection.Methods:(1) Pregnant mice were randomly divided into 4 groups: control group (NS), model group (LPS), prevention group (NAC + LPS) and treatment group (LPS + NAC);(2) Copy preterm birth mouse model of infection. Observe premature time in each group of pregnant mice and premature record is divided into five time periods(0-12h,12-24h,24-36h,36-48h,48h-);(3) Using RT-PCR to detect TLR4,NF-κBp65,TNF-αmRNA expression in the mouse placenta of preterm birth and analysis the relationship between the three and preterm birth with infection, as well as TLR4,NF-κB p65,TNF-αexpression changes after the NAC intervention;(4) Using immunohistochemistry to detect TLR4,NF-κBp65,TNF-αprotein expression in the mouse placenta of preterm birth and TLR4,NF-κB p65,TNF-αexpression after the NAC intervention.Results:(1) Without the occurrence of premature in control group and preterm rate in LPS group is P=0.001, compared with the control group and show that infection model succeeds to set up. Preterm rate in prevention group (P=0.041) significantly decreased, compared with the LPS model group;(2)The NS groups have TLR4,NF-κBp65 and TNF-αexpression at a low level and in LPS model groups expression of the three have significantly increased, compared with the NS groups(P<0.05);(3)In the NAC prevention groups the expression of TLR4 remained unchanged, while the expression of NF-κBp65 and TNF-αhave decreased and the percentage of preterm birth has decreased, compared with the LPS groups(P<0.05);(4)NAC therapy groups were not significant, compared with the LPS groups (P>0.05).Conclusion: Infection model is successful to set up, NAC can reduce the expression of NF-κBp65 and TNF-αin the mouse placenta of premature birth and have a protective effect of premature birth .更多还原