紧密连接结构中相关蛋白在自然妊娠和辅助生殖技术来源胎盘组织中的表达

Expression of tight junction factors in human placental tissues derived from assisted reproductive technology and natural pregnancy

目的 探讨自然妊娠和辅助生殖技术（ART）来源的胎盘组织中紧密连接结构中相关蛋白在胎盘屏障功能中的作用.方法 收集ART来源（ART组10例）和自然妊娠来源（对照组11例）的单胎足月妊娠分娩的胎盘组织.透射电镜观察两组胎盘组织中紧密连接结构的形态学变化；实时荧光定量PCR技术检测紧密连接蛋白（CLDN）1、CLDN4、CLDN5、CLDN8、封闭蛋白（OCLN）以及膜周蛋白（ZO）1的mRNA表达水平,蛋白印迹法检测CLDN4、CLDN8、OCLN蛋白的表达水平.结果 两组胎盘组织均存在清晰的紧密连接结构,定位于合体滋养层细胞细胞间连接近游离面的顶端和血管内皮细胞的细胞连接处；ART组胎盘组织中紧密连接的结构与对照组胎盘组织相似.ART组和对照组胎盘组织中CLDN4mRNA的表达水平分别为0.87±0.17和1.18±0.30,CLDN8 mRNA表达水平分别为3.26±2.32和1.09±0.41,两组分别比较,差异均有统计学意义（P＜0.05）；而ART组和对照组中CLDN1、CLDN5、OCLN以及ZO1 mRNA的表达水平两组间比较,差异均无统计学意义（P＞0.05）.两组胎盘组织中CLDN4、CLDN8、OCLN蛋白表达水平比较,差异也无统计学意义（P＞0.05）.结论 胎盘组织中存在紧密连接结构和相关蛋白的表达；ART来源的胎盘紧密连接结构功能可能潜在地或轻微地受损.

Objective To study the expression of tight junction factors in human placental tissues derived from assisted reproductive technology （ART） and natural pregnancy and its role in placental barrier.Methods Ten placental samples were collected from the women who had undergone ART treatment and 11 placenta were collected from control group.Transmission electron microscope （TEM） examination was utilized to detect the morphology of placental tight junctions.The mRNA of claudin （CLDN） 1,CLDN4,CLDN5,CLDN8,zonula occudens （ZO） 1 was detected by real-time PCR and the protein of CLDN4,CLDN8 and occludin （OCLN） were measured by western blot.Results TEM microscopy results showed that placenta samples derived both ART and control placenta had normal microscopic histological features of tight junctions,localized in the apical part of the syncytium and also between the cell-cell contacts of fetal blood vessel endothelial.The expression level of CLDN4 mRNA were 0.87 ±0.17 in ART group and 1.18 ± 0.30 in control group,respectively.The expression level of CLDN8 mRNA were 3.25 ± 2.32 in ART group and 1.08±0.41 in control group,respectively.The mRNA level of CLDN4 and CLDN8 were significantly differentially expressed in ART derived placenta when compared with control groups.The expression level of CLDN1,CLDN5,OCLN and ZO1 mRNA were 0.49 ± 0.44,0.80 ± 0.20,0.92 ± 0.18 in ART group and 1.09±0.82,1.21 ±0.78,0.80± 0.27 in control group,respectively,in which there were no significant differences between two groups.Western Blot analysis showed the protein levels of tight junctions CLDN4,CLDN8 and OCLN did not differ between groups.Conclusions Tight junction factors were expressed in human placental tissues.Tight junction derived from ATR platenta might have mild dysfunction.