子宫内膜周围淋巴结区素和N-乙酰葡萄糖胺-6-转磺酶表达及其对胚胎着床的影响

Expressions of peripheral lymph node addressin and GlcNAc-6-sulfotransferase in endometrium and their impacts on implantation

目的：探讨人子宫内膜周围淋巴结区素(peripheral lymph node addressin，PNAd)及 N-乙酰葡萄糖胺-6-转磺酶(GlcNkc-6-sulfotransferase，GlcNAc6ST)表达特点及其与胚胎着床的关系。方法：对75例子宫内膜(增生期12例为健康妇女；分泌期63例为不孕症患者，含分泌早期27例，分泌中期36例)采用免疫组织化学法和半定量蛋白免疫印迹法检测 PNAd 的表达，实时荧光定量 PCR 法检测41例分泌期内膜 GlcNAc6ST mRNA。对63例不孕症(男性因素17例，输卵管因素46例)行体外受精-胚胎移植(in vitro fertilization-embryo transfer，IVF-ET)治疗，临床妊娠29例，非妊娠34例，分析两组间 PNAd 和 GlcNAc6ST 表达的差异。结果：(1)PNAd 位于腔上皮和腺上皮细胞胞膜和胞浆，增生期表达较弱，分泌期强表达。PNAd 蛋白免疫印迹呈现至少4条带，对应 Sgp200、CD34、MAdCAM-1，GlyCAM-1四种分子，表达水平呈两两正相关。分泌期 Sgp200、CD34、MAdCAM-1表达高于增生期，差异有统计学意义。各病例均表达 GlcNAc6ST mRNA，与 PNAd 各分子水平无相关。(2)妊娠组 CD34、GIyCAM-1表达高于非妊娠组，差异有统计学意义；两组 GlcNAc6ST mRNA 水平差异无统计学意义。(3)妊娠组与非妊娠组年龄、受精方式、hCG 注射日子宫内膜厚度。

Objective:To investigate the expressions of peripheral lymph node addressin(PNAd)andGlcNAc-6-sulfotransferase(GlcNAc6ST)in endometrium and their impacts on implantation.Methods:PNAd expression in endometrium was examined by immunohistochemistry and Western Blot from 75women(12 from healthy women,in proliferative phase;63 from sterile women,of whom,27 were inearly-secretory and 36 in mid-secretory phase).GlcNAc6ST mRNA was examined by real-time PCR in41 sterile women.The 63 sterile women had underwent ⅣF-ET and were consequently divided into clini-cal pregnant(29 cases)and nonpregnant(34 cases)groups.Results:(1)PNAd localized to the mem-brane and cytoplasm of luminal and glandular epithelia.Staining was patchy and much less intense duringthe proliferative phase than during the secretory phase.In Western Blot of PNAd,four bands appeared,which were Sgp200,CD34,MAdCAM-1,GlyCAM-1 respectively,and each was positively correlatedwith the others significantly.The former three molecular levels were significantly higher during the secre-tory phase as compared with the proliferative phase.Message RNA of GlcNAe6ST was positive in all ca-ses and showed no correlation with any component of PNAd.(2)The expressions of CD34 and GlyCAM-1,but not Sgp200 and MAdCAM-1,were significantly higher in pregnant women than in nonpregnantones.However,the GlcNAc6ST mRNA level did not differ between groups.(3)No significant differ-ence was found in female age,methods of fertilization,thickness of endometrium on day hCG,cumulativeembryo score(CES)and mean score of transferred embryo(MSTE)between the groups.Conclusion:PNAd expression in the human endometrium fluctuates with the menstrual cycle.Elevated CD34 and Gly-CAM-1 during the secretory phase might be stimulative factors for embryo implantation.Defect in PNAdexpression may account for a portion of unexplained infertility.