整合素β3及骨桥蛋白在子宫腺肌病患者子宫内膜中的表达及其意义

Expression of integrin β3 and osteopontin in endometrium of patients with adenomyosis

目的探讨整合素β3及骨桥蛋白（OPN）在子宫腺肌病患者在位及异位子宫内膜中的表达及意义。方法选择2007年1月—2008年7月于北京大学第一医院妇产科因子宫腺肌病行子宫全切除术的子宫腺肌病患者43例，收集其在位子宫内膜（在位内膜组），其中增殖期内膜11例，分泌期内膜32例（其中18例为分泌期中期）；同时收集其异位子宫内膜（异位内膜组）。选择同期因宫颈上皮内瘤变（CIN）Ⅱ-Ⅲ级或单纯浆膜下子宫肌瘤行子宫全切除术的患者41例为对照组，收集其子宫内膜，其中增殖期内膜12例，分泌期内膜29例（其中19例为分泌期中期）。采用免疫组化法和实时荧光定量PCR技术检测各组患者子宫内膜中整合素β3和OPN的蛋白及mRNA表达。结果（1）整合素β3和OPN蛋白主要表达于在位和异位子宫内膜的腺体。子宫内膜中整合素β3和OPN蛋白的表达水平，在位内膜组、异位内膜组、对照组分别为1．6±0．8和1．7±0．7、1．7±0．7和1．8±0．9、2．1±0．9和2．0．±0．9，各组之间分别比较，差异均有统计学意义（P〈0．05）。在位内膜组增殖期子宫内膜中整合素β3和OPN蛋白的表达水平分别为0．8±0．4和0．7±0．3，均低于分泌期（分别为1．8±0．8和1．9±0．8），差异均有统计学意义（P〈0．01）；对照组增殖期子宫内膜中整合素β3和OPN蛋白的表达水平分别为1．0±0．4和1．0±0．4，也均低于分泌期（分别为2．5±0．7和2．5±0．7），差异也均有统计学意义（P=0．000）。在位内膜组分泌期中期子宫内膜中整合素β3和OPN蛋白的表达水平分别为2．0±0．9和2．1±0．8，均低于对照组（分别为2．7±0．5和2．7±0．7），差异均有统计学意义（P〈0．01）。（2）子宫内膜中整合素β3和OPNmRNA的表达水平（以中位数表示），在位内膜组（分别为4．69和4．23）均低于异位内膜组（分别为7．96和14．84）和对照组（分别为13．47和17．40），差异均有统计学意义（P〈0．05）。在位内膜组增殖期子宫内膜中整合素β3和OPNmRNA的表达水平（分别为2．69和3．30）均低于分泌期（分别为5．54和11．40），差异均有统计学意义（P〈0．01）；对照组增殖期子宫内膜中整合素β3和OPNmRNA的表达水平（分别为3．12和4．75）也均低于分泌期（分别为19．94和21．00），差异也均有统计学意义（P=0．000）；在位内膜组分泌期中期子宫内膜中整合素β3和OPNmRNA的表达水平（分别为10．10和14．34）均低于对照组（分别为21．50和24．18），差异均有统计学意义（P〈0．05）。结论整合素β3和OPN在子宫腺肌病患者分泌期中期异位内膜中表达水平较低，可能影响胚胎着床。

Objective To investigate the expression of integrin β3 and osteopontin （OPN） in eutopic and ectopic endometrium of adenomyosis. Methods From January 2007 to July 2008, the endometrium specimens were collected from 43 patients with adenomyosis iundergoing hysterectomy in Peking University First Hospital. Eutopic endometrium were 1 l in proliferative phase and 32 in secretory phase （18 cases in mid-secretory phase） were collected. Ectopic endometriums were also collected. In the mean time, it was chosen 41 cases with pure subserous uterine myoma or cervical intraepithelial neoplasia （CIN） Ⅱ-Ⅲ treated by hysterectomy as controls including 12 endometrium in proliferative phase and 29 endometrium in secretory phase （19 cases in mid-secretory phase）. The expression of Integrin β3 subunit and OPN in the endometrium were assessed by immunohistochemical staining and quantitative real-time polymerase chain reaction. Results （1）Immunohistochemical staining showed that positive staining of integrin β3 and OPN were present predominantly in eutopic and ectopic endometrial glandular epithelium. There was significant different protein expression of integrin β3 and OPN, which were 1.6± 0. 8 and 1.7 ± 0. 7 in eutopic endometrium,1.7 ±0. 7 and 1.8 ±0. 9 in ectopic endometrium,2. 1 ±0. 9 and 2. 0 ±0. 9 in control endometrium （ P 〈 0. 05 ）. The protein expression of integrin β3 and OPN in eutopic endometrium of adenomyosis in the proliferative phase （0. 8 ± 0. 4 and 0. 7 ± 0. 3） were remarkably lower than those of the secretory phase（ 1.8 ± 0. 8 and 1.9 ± 0. 8 ,P 〈 0. 01 ）. The protein expression of integrin β3 and OPN in the endometrium of controls in the proliferative phase （ 1.0 ± 0.4 and 1.0 ± 0. 4 ） were significantly lower than those of the secretory phase （2. 5 ± 0. 7 and 2. 5 ± 0. 7 ） （ P = 0. 000 ）. In the mid-secretory phase, the protein expression of integrin β3 （ 2.0 ± 0. 9 ） and OPN （ 2. 1 ± 0. 8 ） in eutopic endometrium of adenomyosis were significantly lower than that of control endometrium（2. 7 ± 0. 5 and 2.7 ± 0. 7 ） （ P 〈 0. 01 ）. （ 2 ） The mRNA expression level of integrin β3 and OPN in eutopic and ectopic endometrium were assessed by quantitative real-time PCR（ result was shown by median index）. It was observed that integrin β3 mRNA and OPN mRNA were significantly lower in the eutopic endometrium of adenomyosis （4. 69 and 4. 23 ） , when compared with ectopic endometrium （ 7. 96 and 14. 84 ） and controls （ 13.47 and 17.40） （ P 〈 0. 05 ）. Eutopic endometrium had higher mRNA expression of integrin β3 and OPN mRNA in the secretory phase （5.54 and 11.40） than that in the proliferative phase（2. 69 and 3.30） （P 〈0. 01 ）. The mRNA expression level of integrin β3 and OPN of control endometrinm in the proliferative phase （ 3. 12 and 4. 75 ） were significantly lower than that in the secretory phase（ 19. 94 and 21.00, P =0. 000）. The mRNA expression of integrin β3 and OPN were 10. 10 and 14. 34 in the mid-secretory phase, which were significantly lower than 21.50 and 24. 18 in control endometrium （ P 〈 0. 05 ）. Conclusions High expression of integrin β3 and OPN in ectopic endometriurn of adenomyosis may cause endometriotic lesions; abnormal expression of integrin β3 and OPN in the endometrium of adenomyosis during the implantation window may contribute to infertility in some patients.