临床输血与检验 ›› 2016, Vol. 18 ›› Issue (1): 9-13.DOI: 10.3969/j.issn.1671-2587.2016.01.003
张世芬,张颖新,徐军,周继,张军辉,谢红霞
ZHANG Shi-fen, ZHANG Ying-xin, XU Jun, et al
摘要: 目的探讨Th17/Treg细胞在子痫前期(preeclampsia,PE)患者外周血及胎盘组织中表达水平的变化及意义。方法选取PE患者20例及同期正常晚期妊娠20例。应用流式细胞术检测外周血CD4+CD25+ Treg及Th17细胞的表达;荧光定量PCR 检测外周血Treg细胞转录因子Foxp3+mRNA和Th17细胞特异性转录因RORγtmRNA表达;采用免疫组织化学染色,检测蜕膜局部组织中IL17A和Foxp3蛋白的表达。结果PE患者外周血中CD4+CD25+FoxP3+/CD4+比例(2.31士0.54%)明显低于正常产妇(3.86士0.32%)(P<0.05);而子痫前期患者外周血CD4+IL17A+/CD4+比例(2.96士0.28%)明显高于正常产妇(1.61士0.37%)(P<0.05);PE患者外周血Foxp3mRNA表达水平低于对照组,差异有统计学意义(P<0.05),而RORγtmRNA表达高于正常对照组,差异亦有统计学意义(P<0.05);PE患者局部蜕膜组织中IL-17A阳性表达,而正常产妇组局部蜕膜组织相对较低甚至不表达,Foxp3在PE患者的局部蜕膜组织表达相对较低甚至不表达,对照组中阳性表达。结论Th17细胞数目增多,Treg细胞数目减少对于妊娠的维持是不利的,可能是导致子痫前期发生的重要免疫因素。 【Abstract】 Objective To observe the cytokine expressions of Th17 and Treg cells in peripheral blood and the presence of IL-17A and FoxP3 positive cells in decidua with preeclampsia(PE) patients in order to reveal the role of Th17/Treg cells in PE patients. Methods Select 20 cases with PE and 20 cases of healthy women with late pregnancy were selected. Flow cytometry was used to analyze the change of CD4+IL-17A+ and CD4+CD25+ FoxP3+ cells, and qRT-PCR was used to detect the expression of RORγt mRNA of Th17,cells and FoxP3 mRNA of Treg cells in peripheral blood. Decidual tissue was collected in uterus wall placenta and immunohistochemical staining was used to detect the expression of IL-17 and specific transcription factor Foxp3 of decidual CD4+CD25+ regulatory cells. Result Circulating Th17 frequencies (CD4+IL17+/CD4+T cells) were markedly higher in patients with PE (2.96±0.28%) than those with normal gravida with late pregnancy (1.61±0.37%) (P <0.05). However, CD4+CD25+Foxp3+ Tregs in CD4+ cells group in peripheral blood of PE patients(2.31士0.54%) were significantly lower than those in normal gravidas (3.86±0.32%, P<0.05). Simultaneously, Treg cell frequencies of normal early pregnant women were increased compared with control(P<0.05). The expression of RORγt mRNA in peripheral blood of PE patients was significantly higher than that in normal control group, while the expression of FoxP3 mRNA in peripheral blood of PE patients was markedly lower than that in normal control group(P <0.05). Additionally, the positive expression of IL-17A in decidua of PE patients was noted, while slight expression was seen in normal pregnancy. Conclusion The number of Th17 cells was increased whereas Treg cells were decreased in PE patients. The imbalance of Th17 and Treg cells may play an important role in the immune system in the pathophysiology of preeclampsia.
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