17β雌二醇对子宫腺肌病患者子宫内膜-肌层交界区平滑肌细胞内游离Ca^2＋浓度的调节作用及机制研究  

Mechanism of 17β-estrogen on intracellular free calcium regulation in smooth muscle cells at the endometrial-myometrial interface in uteri with adenomyosis

作　　者：汪沙[1] 段华[1] 张颖[1] 王丽平[1] 张恒辉[3] 李国力[2] 

机构地区：[1]首都医科大学附属北京妇产医院妇科微创中心,100006 [2]首都医科大学附属北京地坛医院传染病研究所,100006 [3]北京大学人民医院肝病研究所

出　　处：《中华妇产科杂志》2015年第7期510-515,共6页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金（81270680）;北京市医院管理局重点医学专业发展计划（ZYLX201406）;北京市自然科学基金（7142056）

摘　　要：目的探讨17β雌二醇（17β-E2）对子宫腺肌病患者子宫内膜-肌层交界区（EMI）平滑肌细胞内游离Ca^2＋浓度（[Ca^2＋]i）的调节作用及可能机制。方法选择2011年9月至2012年12月在首都医科大学附属北京妇产医院妇科微创中心因子宫腺肌病行子宫全切除术的患者28例为腺肌病组，选择同期因子宫颈上皮内瘤变（CIN）Ⅲ行子宫全切除术者31例为对照组，对两组患者的EMI平滑肌细胞进行分离及原代培养。应用Ca^2＋荧光探针Fluo-4／AM负载平滑肌细胞，采用细胞外钙阻断实验、细胞内钙释放阻断实验、细胞膜钙通道阻断实验、细胞膜钙激活钾通道阻断实验阻断引起[Ca^2＋]i变化的各个环节，利用激光共聚焦显微镜观察17β-E2诱导后的两组患者EMI平滑肌细胞内[Ca^2＋]i的变化，其变化程度以ΔF[Ca^2＋]i表示。结果（1）在常规有Ca^2＋培养基中，17β-E2诱导后腺肌病组和对照组EMI平滑肌细胞内Ca^2＋荧光强度均明显高于其17β-E2诱导前，腺肌病组ΔF[Ca^2＋]i为384±26，对照组ΔF[Ca^2＋]i为235±20，两组比较，差异有统计学意义（P=0．001）；细胞外钙阻断实验显示，细胞外无Ca^2＋时，腺肌病组ΔF[Ca^2＋]i为207±17，对照组ΔF[Ca^2＋]i为221±19，两组比较，差异无统计学意义（P=0．731）。腺肌病组ΔF[Ca^2＋]i在细胞外有Ca^2＋时明显高于无Ca^2＋时，差异有统计学意义（P=0．000）；而对照组两者比较，差异无统计学意义（P=0．060）。（2）细胞内钙释放阻断实验显示，阻断肌质网钙库上三磷酸肌醇（IP3）敏感的钙释放通道、肌质网利阿诺定（Ryanodine）受体（RyR）钙释放通道、肌质网钙泵释放后，腺肌病组和对照组ΔF[Ca^2＋]i分别与其阻断前相比均明显降低（P〈0．05），且腺肌病组ΔF[Ca^2＋]i均明显高于对照组（P〈0．05）。（3）细胞膜钙通道阻断实验显示，阻断L-型钙通道后，腺肌�Objective To investigate the regulation mechanism of estrogen on the free calcium of smooth muscle cells at the endometrial-myometrial interface （EMI） in uteri with adenomyosis. Methods From September 2011 to November 2012, 59 uterine myometrial specimens were obtained from 59 cases underwent hysterectomy, including 28 adenomyosis patients as adenomyosis （ADS） group and 31 patients with cervical intraepithelial neoplasia Ⅲ as control group. EMI smooth muscle cells were cultured and loaded with calcium ion fluorescent probe fluo-4/AM. After treated with trisphosphate （IP3） receptor antagonist, blocker of sarcoplasmic reticulum calcium-adenosine triphosphate （ATP）, depleted agent of the ryanodine receptor-operated Ca^2＋, inhibitor of L-type calcium channel, inhibitor of Na^＋-Ca^2＋ exchanger, the labeled cells were stimulated with estrogen. The changes of intracellular Ca^2＋ fluorescence intensity were detected by laser scanning microscopy. The changes of intracellular Ca^2＋ concentration was indicated by ΔF[Ca^2＋]i. Results （1） Under normal calcium conditions, after the stimulation of estrogen, iutraeellular Ca^2＋ fluorescence intensity in ADS group and control group both increased than those without estrogen. The ΔF[Ca^2＋]i in ADS group was 384±26, and in the control group ΔF[Ca^2＋]i was 235±20. The ΔF[Ca^2＋]i in ADS group was higher than that in the control （P〈O.O1）. Without calcium conditions, the ΔF[Ca^2＋]i in ADS group was 207± 17, and in the control group ΔF[Ca^2＋]i was 221 ± 19. The ΔF[Ca^2＋]i in ADS group was almost the same with the increase in the control （P=0.731）. The ΔF[Ca^2＋]i in ADS group was significantly decreased compared with the calcium condition （P〈0.01）. However, there was no significant difference in the control between with and without calcium conditions （P=0.060）. （2） After treated with IP3 receptor antagonist, blocker of sarcoplasmic reticulum caleium-ATP, depleted agent of the ryanodine receptor-operated

关 键 词：子宫腺肌病 雌二醇 子宫内膜 子宫肌层 肌细胞 平滑肌 缝隙接合部 钙 

分 类 号：R711.74[医药卫生—妇产科学]