胎盘粘连植入组织和人胎盘滋养层细胞中VCAM-1的表达及其与侵入性胎盘疾病的相关性

目的探讨胎盘粘连植入组织和人胎盘滋养层细胞中血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)的表达及其与侵入性胎盘疾病的相关性。方法采用酶联免疫吸附试验检测2017年9月至2018年9月期间我院收治的20例产妇静脉血中可溶性VCAM-1(sVCAM-1)的表达水平,根据sVCAM-1的平均值将另外随机选取的200例产妇分为高表达组和低表达组,计算两组侵入性胎盘发生率。采用免疫组化染色、实时定量PCR和Western blot检测40例侵入性胎盘组织和40例正常胎盘组织中的VCAM-1、TNF-α、NF-κB p65和IκBα表达。应用20 ng/mL的TNF-α处理人胎盘滋养层HTR8/SVneo细胞系后检测细胞中VCAM-1、TNF-α、NF-κB p65和IκBα的表达。结果20例健康产妇静脉血中的VCAM-1平均水平为36.32±3.25 ng/mL。高表达组的侵入性胎盘发生率(6.98%)显著高于低表达组(0.64%)(χ^2=0.922,P=0.009)。实时定量RT-PCR和Western blot结果表明,与对照组相比,侵入性胎盘组VCAM-1的mRNA和蛋白表达水平显著升高。免疫染色显示VCAM-1蛋白主要在滋养细胞的细胞质和膜中表达,侵入性胎盘组的VCAM-1阳性率(72.50%)显著高于对照组(17.50%)(Z=-5.063,P<0.001)。与对照组相比,侵入性胎盘组的TNF-α和NF-κB p65表达水平显著升高,而IκBα表达水平显著降低(P<0.05)。与未用TNF-α处理(0 ng/mL)的HTR8/SVneo细胞相比,应用20 ng/mL浓度的TNF-α处理细胞1周后,细胞中的NF-κB p65和VCAM-1表达水平显著升高,而IκBα表达水平显著降低(P<0.05)。结论VCAM-1在侵入性胎盘产妇血液和胎盘组织中均为高表达模式。VCAM-1的活化至少部分依赖于TNF-α、NF-κB信号通路的激活,从而诱导其在滋养细胞中高表达并引起过度侵袭,导致侵入性胎盘的发生。

The Expression of VCAM-1 in Placenta Adhesion Implants and Human Placental Trophoblast Cells and Its Correlation with Invasive Placental Disease

Objective To investigate the expression of vascular cell adhesion molecule-1(VCAM-1)in placenta adhesion implants and human placental trophoblast cells and its correlation with invasive placental disease.Methods The expression of soluble VCAM-1(sVCAM-1)in 20 cases of venous blood in pregnant women admitted to our hospital from September,2017 to September,2018 was measured by enzyme-linked immunosorbent assay(ELISA).According to the average value of sVCAM-1,another 200 cases of pregnant women were randomly divided into the high expression group and low expression group,and the incidence of invasive placenta was calculated.Immunohistochemical staining,real-time quantitative PCR and western blot were used to detect the expressions of VCAM-1,TNF-α,NF-κB p65 and IκBαin 40 cases of invasive placenta and 40 cases of normal placenta.The expressions of VCAM-1,TNF-α,NF-κB p65 and IκBαin the human placental trophoblast HTR8/SVneo cell line were detected after the treatment with 20 ng/mL TNF-α.Results The mean level of VCAM-1 in venous blood of 20 healthy women was 36.32±3.25 ng/mL.The incidence of invasive placenta(6.98%)in the high expression group was significantly higher than that in the low expression group(0.64%)(χ^2=0.922,P=0.009).Real-time quantitative RT-PCR and western blot results showed that the mRNA and protein expression levels of VCAM-1 in the invasive placenta group were significantly higher than those in the control group.Immunostaining showed that VCAM-1 protein was mainly expressed in the cytoplasm and membrane of trophoblast cells.The positive rate of VCAM-1 in the invasive placenta group(72.50%)was significantly higher than that in the control group(17.50%)(Z=-5.063,P<0.001).Compared with the control group,the expression levels of TNF-αand NF-κB p65 in the invasive placenta group were significantly increased,while the expression level of IκBαwas significantly decreased(P<0.05).Compared to HTR8/SVneo cells treated with TNF-α(0 ng/mL),NF-κB p65 and VCAM-1 expression levels in HTR8/SVneo cells treated with TNF-αat a concentration of 20 ng/mL for 1 week were significantly elevated,while IκBαexpression levels were significantly decreased(P<0.05).Conclusion VCAM-1 shows a high expression pattern in maternal blood and placental tissues of invasive placenta.Activation of VCAM-1 depends,at least in part,on the activation of the TNF-α/NF-κB signaling pathway,thereby inducing its high expression in trophoblast cells and causing excessive invasion,further leading to the development of invasive placenta.