Title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游离Ca2+调节模式的初步研究
Other TitlesPreliminary study of estrogen effects on calcium free smooth muscle cells at the endometrialmyometrial interface in uteri with adenomyosis
Authors王丽平
汪沙
张颖
王永军
张恒辉
常亚楠
李国力
段华
Affiliation100006,首都医科大学附属北京妇产医院妇科微创中心
北京大学人民医院肝病研究所
100006,首都医科大学附属北京地坛医院传染病研究所
Keywords雌二醇
子宫内膜异位症
子宫内膜
子宫肌层

Estradiol
Endometriosis
Endometrium
Myometrium
Calcium
Issue Date2012
Publisher中华妇产科杂志
Citation中华妇产科杂志.2012,47,(5),351-354.
Abstract目的 观察雌二醇对子宫腺肌病患者子宫内膜-肌层交界区(EMI)平滑肌细胞游离Ca2+浓度的影响,并探索其作用模式.方法 选择2011年3月至10月在首都医科大学附属北京妇产医院因子宫腺肌病行子宫全切除术的患者16例,其中子宫内膜增殖期9例、分泌期7例.取其EMI平滑肌组织进行原代细胞培养,以Ca2+荧光探针负载平滑肌细胞,用激光共聚焦显微镜观察1 ×102、1×103、1×104、1×105 pmol/L浓度的雌二醇作用后平滑肌细胞内Ca2+浓度(以Ca2+荧光强度表示)的变化,选择最佳浓度雌二醇.以最佳浓度雌二醇分别联合17β雌二醇-牛血清白蛋白复合物(17β-E2-BSA)、ER拮抗剂——氟维司群(ICI182780)、蛋白合成抑制剂——放线菌酮(CHX)、G蛋白活化抑制剂——百日咳毒素(PTX)处理者分别为17β-E2-BSA组.ICI182780组、CHX组和PTX组,以仅用最佳浓度雌二醇处理者为上述各组的相应对照组,观察雌二醇作用后增殖期、分泌期EMI平滑肌细胞以及各组EMI平滑肌细胞内Ca2+荧光强度的变化.结果 (1)EMI平滑肌细胞培养24h后即贴壁生长,状态良好.(2)1×102 ~1 × 105 pmol/L浓度的雌二醇均能在1 min内引起细胞内Ca2荧光强度迅速升高,分别为275±16、449±18、615±36、641±47,其中1 × 104 pmol/L与1×105 pmol/L浓度的雌二醇作用下细胞中Ca2+荧光强度增幅最明显,但两者比较,差异无统计学意义(P>0.05),因此选择1×104 pmol/L为最佳雌二醇浓度.(3)1×104 pmol/L浓度雌二醇作用于增殖期与分泌期EMI平滑肌细胞,两者Ca2+荧光强度比较,差异无统计学意义(P>0.05);17β-E2-BSA组与CHX组细胞内Ca2+荧光强度分别为646±32和602±31,与各自相应对照组(分别为513±26和617±35)比较,差异均无统计学意义(P>0.05);而PTX组与ICD82780组分别为188±20和302±11,与各白相应对照组(分别为632±33和635±24)比较,差异均有统计学意义(P<0.01).结论 雌二醇对子宫腺肌病患者EMI平滑肌细胞内游离Ca2+浓度的调节符合膜受体介导的非基因转录作用模式.
Objective To investigate the effect and mechanism of estrodial (E2) on intracellular free calcium in the endometrial-myometrial interface (EMI) smooth muscle cells from uteri with adenomyosis.Methods From March 2011 to October 2011,16 uterus specimens were collected from patients with adenomyosis undergoing hysterectomy in Beijing Obstetrics and Gynecology Hospital,which included 9 proliferative endometrium and 7 secretory endometrium.EMI smooth muscle cells from the uterus were cultured and loaded with calcium ion ( Ca2 + ) fluorescent probe fluo-4/AM.The labeled cells were stimulated with the various concentration of E2 ( 1 × 102,1 × 103,1 × 104,1 × 105 pmol/L,respectively),then the changes of intracellular Ca2+ fluorescence intensity were measured by laser scanning microscopy.The most suitable concentration of E2 was selected,and the reaction difference between the EMI smooth muscle cells of two menstrual phases were also investigated; The changes of intracellular Ca2 + fluorescence intensity were detected proliferative and secretory smooth muscle cells in E2 conjugated to bovine serum albumin (17β-E2-BSA) group,cycloheximide (CHX) group,fulvestrant (ICI182780) group and pertussis toxin (PTX) group.Results ( 1 ) The cell viability of primary cultured EMI smooth muscle cells was well at 24 hours culture.(2) 1 × 102 - 1 × 105 pmol/L E2 can rapidly increase the intracellular Ca2+ fluorescence intensity within 1 min ( P < 0.01 ) ;The increased amplitudes caused by 1 × 104 pmol/L and 1 × 105 pmoL/L E2 were the most significant,but there was no significant difference between them (P >0.05).1 × 104 pmol/L was the most suitable concentration.( 3 ) With the 1 × 104 pmol/L E2,the Ca2+ fluorescence intensity changes showed no significant difference between the EMI smooth muscle cells from the proliferative phase and secretory phase uterus (P > 0.05 ).The Ca2+ fluorescence intensity changes were 646 ± 32 in 17β-E2-BSA group and 602 ±31 in CHX group,when compared with 513 ±26 and 617 ±35 in respective control group,no significant difference was observed (P > 0.05 ).The increased amplitude of 188 ± 20 in the PTX group and 302 ± 11 in ICI182780 group exhibited significant difference with 632 ± 33 and 635 ± 24 in respective control group ( P < 0.01 ).Conclusion E2 could increase the intracellular Ca2 + of EMI through a membrane receptor dependent and nongenomic mechanism of action.
URIhttp://hdl.handle.net/20.500.11897/287716
ISSN0529-567X
DOI10.3760/cma.j.issn.0529-567x.2012.05.008
Indexed中文核心期刊要目总览(PKU)
中国科技核心期刊(ISTIC)
中国科学引文数据库(CSCD)
Appears in Collections:人民医院

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