CRH Activation of Different Signaling Pathways Results in Differential Calcium Signaling in Human Pregnant Myometrium before and during Labor
CRH Activation of Different Signaling Pathways Results in Differential Calcium Signaling in Human Pregnant Myometrium before and during Labor
复制标题
不同信号通路的 CRH 激活导致人类妊娠子宫肌层在分娩前和分娩过程中钙信号传导存在差异。
DOI：
10.1210/jc.2011-3383
复制
发表时间：
2012-10-01
影响因子：
5.8
通讯作者：
Ni, Xin
中科院分区：
文献类型：
--
作者：
You, Xingji;Gao, Lu;Ni, Xin
背景
我们之前的研究表明，CRH 对临产前后的人类子宫收缩力有不同的影响。细胞内 Ca2+ 浓度 ([Ca2+]i) 动员在子宫收缩的控制中起着重要作用。
目标
我们的目的是研究 CRH 对临产和非临产子宫肌细胞 [Ca2+]i 稳态的影响，并确定 CRH 参与 [Ca2+]i 调节的后续信号传导。
设计
子宫肌层组织取自足月分娩或未足月分娩的孕妇。 [Ca2+]i 通过 Ca2+ 成像系统使用荧光染料 fura-2-乙酰氧基甲酯测定。使用蛋白质印迹分析、ELISA 和 RIA 来确定 CRH 诱导的信号通路。
结果
CRH 在分娩细胞中诱导 Ca2+ 瞬变，该现象被 CRH 受体 1 型 (CRHR1) 拮抗剂安塔拉明 (antalarmin) 阻断。 CRHR1 敲除削弱了 CRH 的这种作用。 CRH 激活 Gi 蛋白，减少 cAMP 产生，并诱导磷酸化磷脂酶 C-β3 和肌醇-1,4,5-三磷酸的产生。磷脂酶 C 和肌醇 1,4,5-三磷酸受体抑制剂可阻断分娩细胞中 CRH 诱导的 Ca2+ 瞬变。 CRH 不诱导非临产细胞中的 Ca2+ 瞬变，而 antalarmin 则诱导 Ca2+ 瞬变。 CRHR1 的敲低削弱了 antalarmin 的作用。 CRH 作用于 CRHR1，激活非临产细胞中的 Gs。 Forskolin 阻断 antalarmin 诱导的 Ca2+ 瞬变。
结论
CRH 作用于 CRHR1，在临产前后激活不同的信号通路，从而导致响应 CRH 的差异钙信号传导。 CRHR1 的信号通路可能作为开发新的早产治疗策略的靶点。
CONTEXT
Our previous study has demonstrated that CRH has differential effects on human uterine contractility before and after onset of labor. Intracellular Ca2+ concentration ([Ca2+]i) mobilization plays an important role in the control of uterine contraction.
OBJECTIVE
Our objective was to investigate the effects of CRH on [Ca2+]i homeostasis in laboring and nonlaboring myometrial cells and determine subsequent signaling involved in [Ca2+]i regulation by CRH.
DESIGN
The myometrial tissues were obtained from pregnant women who were undergoing or not undergoing labor at term. [Ca2+]i was determined by Ca2+ imaging system using the fluorescent dye fura-2-acetoxymethyl ester. Western blot analysis, ELISA, and RIA were used to determine the signaling pathways induced by CRH.
RESULTS
CRH induced Ca2+ transient in laboring cells, which was blocked by CRH receptor type 1 (CRHR1) antagonist antalarmin. CRHR1 knockdown impaired this effect of CRH. CRH activated Gi protein, decreased cAMP production, and induced phosphorylated phospholipase C-β3 and inositol-1,4,5-triphosphate production. Phospholipase C and inositol-1,4,5-triphosphate receptor inhibitors blocked the CRH-induced Ca2+ transient in laboring cells. CRH did not induce whereas antalarmin induced the Ca2+ transient in nonlaboring cells. Knockdown of CRHR1 impaired the effect of antalarmin. CRH acted on CRHR1 to activate Gs in nonlaboring cells. Forskolin blocked antalarmin-induced Ca2+ transient.
CONCLUSIONS
CRH acts on CRHR1 to activate different signaling pathways before and after onset of labor, thereby resulting in differential calcium signaling in response to CRH. The signaling pathways of CRHR1 might serve as a target for the development of new therapeutic strategies for preterm birth.