Induction of cyclooxygenase-2 expression in human myometrial smooth muscle cells by interleukin-1β:: involvement of p38 mitogen-activated protein kinase
Induction of cyclooxygenase-2 expression in human myometrial smooth muscle cells by interleukin-1β:: involvement of p38 mitogen-activated protein kinase
复制标题
DOI：
10.1111/j.1469-7793.1999.00399.x
复制
发表时间：
1999-10-15
影响因子：
5.5
通讯作者：
Mann, GE
中科院分区：
文献类型：
--
作者：
Bartlett, SR;Sawdy, R;Mann, GE
1. 将培养中的人子宫平滑肌细胞（HMSMC）暴露于重组人白介素-1β（IL-1β，10 ng ml(-1)）中1至24小时。环氧合酶-2 (COX-2) mRNA 和蛋白被快速诱导，表达持续 24 h。放线菌酮(10 μg ml(-1) 6 h)阻断IL-1β诱导的COX-2蛋白表达和超诱导COX-2 mRNA表达。地塞米松(1μm，6h)阻断COX-2 mRNA和蛋白的诱导。3． IL-1β诱导的COX-2表达伴随着向培养基中的前列腺素E-2释放增加3倍。4． IL-1 beta 诱导 HMSMC 中 p42/4 1 和 p38 丝裂原激活蛋白激酶 (MAPK) 的瞬时（5-30 分钟）激活。 p38 MAPK 的活性通过其底物 MAP 激酶激活蛋白激酶-2 (MAPKAP 激酶-2) 的凝胶活性进行监测。 p38 MAPK 抑制剂 SE 203580（10 μm，5-30 分钟）可阻止 MAPKAP 激酶-2 活性的诱导。5． IL-1β刺激6小时后检测到的COX-2蛋白表达被SE 203580 (10μm)阻断。将HMSMC暴露于10 ng ml(-1) IL-1 beta仅30分钟，在6小时培养物中诱导COX-2蛋白表达水平与暴露于细胞因子6小时的细胞中检测到的水平相似。6。仅在 IL-1 β 刺激的前 30 分钟内将细胞暴露于 SB 203580 (10 μm)，可有效阻断培养 6 小时后测定的 COX-2 蛋白表达。7．这项研究已经确定，p38 MAPK 级联的瞬时激活参与了人子宫平滑肌细胞中 IL-1 β 刺激的 COX-2 表达。 HMSMC 中 IL-1β 诱导 COX-2 为以下假设提供了支持：子宫肌层中的自分泌前列腺素信号传导（由宫内细胞因子浓度升高引发）在分娩期间调节子宫肌层收缩力中发挥作用。
1. Human myometrial smooth muscle cells (HMSMCs) in culture were exposed to recombinant human interleukin-1 beta (IL-1 beta, 10 ng ml(-1)) for 1 to 24 h. Cyclooxygenase-2 (COX-2) mRNA and protein were rapidly induced, with espression sustained at 24 h.2. Cycloheximide (10 mu g ml(-1) 6 h) blocked IL-1 beta-induced COX-2 protein expression and super-induced COX-2 mRNA expression. Induction of COX-2 mRNA and protein was blocked by dexamethasone (1 mu m, 6 h).3. IL-1 beta-induced COX-2 expression was accompanied by a 3-fold increase of prostaglandin E-2 release into the culture medium.4. IL-1 beta induced a transient (5-30 min) activation of p42/4 1 and p38 mitogen-activated protein kinase (MAPK) enzymes in HMSMCs. Activity of p38 MAPK was monitored by in gel activity of its substrate MAP kinase-activated protein kinase-2 (MAPKAP kinase-2). Induction of MAPKAP kinase-2 activity was prevented by the p38 MAPK inhibitor SE 203580 (10 mu m, 5-30 min).5. COX-2 protein expression detected after 6 h IL-1 beta stimulation was blocked by SE 203580 (10 mu m). Exposure of HMSMCs to 10 ng ml(-1) IL-1 beta for only 30 min induced a level of COX-2 protein expression at 6 h culture similar to that detected in cells exposed to the cytokine for 6 h.6. Exposure of cells to SB 203580 (10 mu m) during only the first 30 min of IL-1 beta stimulation was effective in blocking COX-2 protein expression assayed after 6 h in culture.7. This study has established that a transient activation of the p38 MAPK cascade is involved in IL-1 beta-stimulated COX-2 expression in human myometrial smooth muscle cells. Induction of COX-2 by IL-1 beta in HMSMCs provides support for the hypothesis that autocrine prostaglandin signalling in the myometrium, initiated by elevated intrauterine cytokine concentrations, plays a role in regulating myometrial contractility during labour.