重度子痫前期蜕膜NK细胞Tim-3的表达及其相关作用  

The expression and role of decidual NK cell Tim-3 in severe preeclampsia

作　　者：王雪 张恂 WANG Xue;ZHANG Xun(Department of Obstetrics and Gynecology,Sichuan Women’s and Children’s Hospital,Chengdu 610045,China;Department of Obstetrics and Gynecology,Sichuan People’s Hospital,Sichuan Academy of Medical Sciences,Chengdu 610072,China)

机构地区：[1]四川省妇幼保健院·四川省妇女儿童医院妇产科,成都610045 [2]四川省医学科学院·四川省人民医院妇产科,成都610072

出　　处：《免疫学杂志》2021年第2期145-152,共8页Immunological Journal

摘　　要：目的探讨重度子痫前期Tim-3+CD56^+CD16^-NK细胞的表达及其对滋养细胞侵袭及血管内皮细胞成管能力的影响。方法分离重度子痫前期患者(sPE组)及正常妊娠孕产妇(Normal组)的胎盘蜕膜组织中单个核细胞,细胞流式检测蜕膜NK细胞(decidual NK cell,dNK)细胞所占比例;免疫磁珠分选CD56^+CD16^-NK细胞,细胞流式及RT-PCR实验检测Tim-3在其中的表达;将分选出的CD56^+CD16^-NK细胞分为3组:正常对照(Control)组,即来源于正常妊娠胎盘蜕膜组织中的dNK细胞;重度子痫前期(sPE)组,即来源于重度子痫前期胎盘蜕膜组织中的dNK细胞;Tim-3封闭(Tim-3-Fc)组,即加入Tim-3-Fc融合蛋白进行处理的正常妊娠胎盘蜕膜组织中的dNK细胞。采用ELISA、Transwell侵袭及小管形成实验分别检测上述3组细胞上清液中TNF-α、IL-6、IL-8、VEGF及IFN-γ的表达及其对HTR8侵袭能力及血管内皮细胞成管能力的影响。结果与Control组相比,sPE组中CD56^+CD16^-NK细胞占蜕膜组织单个核细胞比例显著减少(P<0.05);免疫磁珠可分选出纯度达90%以上CD56^+CD16^-NK细胞,且sPE组中Tim-3的表达丰度及Tim-3 mRNA表达水平均较Control组显著降低(P<0.05);ELISA结果显示与Control组相比,sPE组与Tim-3-Fc组dNK细胞上清液中TNF-α、IFN-γ表达均显著增加(P<0.05),而IL-6、IL-8、VEGF表达均明显降低(P<0.05);Transwell侵袭实验及成管实验结果表明,sPE组与Tim-3-Fc组HTR8细胞侵袭能力及HUVEC成管能力较Control组显著降低(P<0.05),但Tim-3-Fc组HTR8细胞侵袭能力及HUVEC成管能力较sPE组明显增强(P<0.05)。结论重度子痫前期患者Tim-3+CD56^+CD16^-NK的表达显著降低,其可能通过损害蜕膜NK细胞的分泌功能来抑制滋养细胞的侵袭及血管内皮细胞的成管能力,从而促进子痫前期的发展。This study was designed to investigate the expression of Tim-3+CD56^+CD16^-NK cells in severe preeclampsia and its effect on the invasion of trophoblasts and the tube formation capacity of vascular endothelial cells.Mononuclear cells were isolated from the placental decidua of patients with severe preeclampsia(sPE group)and normal pregnant women(normal group).CD56^+CD16^-NK(dNK)cells were separated by immunomagnetic beads,and the expression of Tim-3 in CD56^+CD16^-NK cells was detected by flow cytometry and RT-PCR.The CD56^+CD16^-NK cells were divided into three groups:control group(derived from normal decidual tissue of placenta),severe preeclampsia(sPE)group(derived from the decidua of sPE placenta),and Tim-3-Fc group(derived from the decidua of normal pregnancy placenta treated with Tim-3-Fc fusion protein).The expression levels of TNF-α,IL-6,IL-8,VEGF and IFN-γin the supernatant of the three groups were detected by ELISA.dNK cells were co-cultured with HTR8 to perform the invasion assay,while the supernatants of dNK cells were co-cultured with HUVEC to perform the tube formation assay.Data showed that compared with control group,the proportion of CD56^+CD16^-NK cells in decidual mononuclear cells of sPE group was significantly decreased(P<0.05).The purity of CD56^+CD16^-NK cells was more than 90%.The expression of Tim-3 and Tim-3 mRNA in sPE group were significantly lower than those in control group(P<0.05).And the results of ELISA showed that compared with control group,the expression of TNF-αand IFN-γin supernatant of dNK cells in sPE group and Tim-3-Fc group were significantly increased(P<0.05),while the expressions of IL-6,IL-8 and VEGF were significantly decreased(P<0.05).The results of Transwell invasion and tube formation assay showed that pro-invasion and proangiogenesis of dNK cells in sPE group and Tim-3-Fc group were significantly lower than those in control group(P<0.05),and the pro-invasion and pro-angiogenesis of dNK cells in Tim-3-Fc group were significantly higher than those in sPE group(P

关 键 词：重度子痫前期 蜕膜NK细胞 TIM-3 滋养细胞 血管内皮细胞 

分 类 号：R344.04[医药卫生—基础医学]