促性腺激素释放激素激动剂对子宫内膜异位症不孕患者在位内膜一氧化氮合酶表达的影响

目的探讨一氧化氮合酶(NOS)在子宫内膜异位症(内异症)不孕患者在位子宫内膜(在位内膜)中的表达及促性腺激素释放激素激动剂(GnRHa)对其表达的影响。方法采用免疫组化方法及蛋白印迹方法,对30例Ⅲ～Ⅳ期内异症不孕患者(内异症组)及19例有正常生育史的宫颈原位癌患者(对照组)的在位内膜进行内皮型NOS(eNOS)及诱生型NOS(iNOS)表达的定位及半定量检测,内异症组中18例行GnRHa治疗(治疗组);采用电化学发光法,同时测定患者血清雌二醇及孕酮水平。结果eNOS在子宫内膜的腺上皮、腔上皮及血管内皮细胞中均有表达;iNOS仅在子宫内膜的腺上皮和间质细胞中微弱表达。eNOS在增生期早、中、晚期和分泌期早、中、晚期在位内膜中的相对表达水平,内异症组分别为0.30±0.04,0.40±0.03,0.49±0.03,0.43±0.04,0.55±0.04和0.48±0.03,治疗组分别为0.22±0.03,0.37±0.03,0.45±0.04,0.35±0.05,0.50±0.03和0.41±0.00,对照组分别为0.21±0.03,0.33±0.03,0.45±0.04,0.40±0.03,0.47±0.05和0.41±0.03。在整个月经周期中,内异症组患者在位内膜eNOS相对表达水平持续性高于对照组患者,但仅在增生期早、中期及分泌期中、晚期两组比较,差异有统计学意义(P<0.05);与内异症组比较,治疗组在位内膜eNOS相对表达水平均有所下降,但仅在增生期早期及分泌期早、中期比较,差异有统计学意义(P<0.05)。3组子宫内膜中均未检出iNOS的表达。血清雌二醇或孕酮水平与子宫内膜eNOS表达水平均呈正相关(P<0.01)。结论eNOS在内异症患者在位内膜中的高表达,可能与其发病及其所致的不孕有关,GnRHa治疗可降低内异症患者在位内膜eNOS的表达。

Influences of gonadotropin releasing hormone agonist treatment on nitric oxide synthase expression in women with endometriosis and infertility

OBJECTIVE: To explore the expression of endothelial and inducible nitric oxide synthase (eNOS and iNOS) in the eutopic endometria of patients with endometriosis and infertility, and to detect the effect of gonadotropin releasing hormone agonist (GnRH-a) therapy on their expressions. METHODS: Immunohistochemistry and Western immunoblot assay were used to screen eNOS and iNOS expression in endometria of patients with stage III to IV endometriosis and infertility without (30 cases, endometriosis group) and with GnRH-a treatment (18 cases, treatment group), and of 19 patients with carcinoma in situ of the cervix (control group). Using electrochemiluminescence immunoassay, serum estradiol (E(2)) or progesterone (P) concentrations were assayed. RESULTS: eNOS was localized predominantly to the endometrial glandular epithelium, luminal epithelium, and microvascular endothelium. iNOS staining was light, and it, when present, was predominantly found in glandular epithelium and stromal cells. By Western immunoblot analysis, iNOS was not detected. During the early-, mid-, and late-proliferative phases, and the early-, mid-, and late-secretory phases, endometrial eNOS protein relative levels were 0.30 +/- 0.04, 0.40 +/- 0.03, 0.49 +/- 0.03, 0.43 +/- 0.04, 0.55 +/- 0.04 and 0.48 +/- 0.03 in endometriosis group, 0.22 +/- 0.03, 0.37 +/- 0.03, 0.45 +/- 0.04, 0.35 +/- 0.05, 0.50 +/- 0.03 and 0.41 +/- 0.00 in treatment group, and 0.21 +/- 0.03, 0.33 +/- 0.03, 0.45 +/- 0.04, 0.40 +/- 0.03, 0.47 +/- 0.05 and 0.41 +/- 0.03 in control group, respectively. Eutopic endometria in endometriosis group showed higher levels of eNOS protein than that of the control group, but a significant difference was found only in early-, mid-proliferative phases and mid-, late-secretory phases (P < 0.05). Compared with endometriosis group, endometrial eNOS levels in treatment group were reduced, but a significant difference was found only in early proliferative phase and early-, mid-secretory phases (P < 0.05). A significant positive correlation was found between serum E(2) or P concentrations and endometrial expression of eNOS (P < 0.01). CONCLUSIONS: The higher endometrial expression of eNOS in patients with stage III to IV endometriosis may be related to the pathogenesis of endometriosis and the associated subfertility. The expression of eNOS is markedly reduced by administration of GnRH-a.