Crosstalk between monocytes and myometrial smooth muscle in culture generates synergistic pro-inflammatory cytokine production and enhances myocyte contraction, with effects opposed by progesterone.
Crosstalk between monocytes and myometrial smooth muscle in culture generates synergistic pro-inflammatory cytokine production and enhances myocyte contraction, with effects opposed by progesterone.
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DOI：
10.1093/molehr/gav027
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发表时间：
2015-08
影响因子：
4
通讯作者：
Norman JE
中科院分区：
文献类型：
--
作者：
Rajagopal SP;Hutchinson JL;Dorward DA;Rossi AG;Norman JE
足月和早产的特征都是巨噬细胞和中性粒细胞涌入子宫肌层和宫颈，同时外周血单核细胞激活增加。感染和炎症与早产(PTL)的病理密切相关，孕酮被认为是预防或治疗早产的一种有前途的候选药物。在这项研究中，我们研究了单核细胞单独和在体内对用于触发PTL的TLR4激动剂LPS的反应对子宫肌层平滑肌细胞炎性细胞因子产生的影响。我们还研究了单核细胞对心肌细胞收缩的影响。从足月妊娠妇女外周血中分离出单核细胞，单独培养或与PHM1-41子宫肌层细胞共培养24小时，第三组实验分离培养PHM1-41肌细胞24小时。细胞因子的分泌采用双抗体夹心ELISA法或多重测定法。单核细胞和心肌细胞共培养可协同分泌促炎细胞因子和趋化因子IL-6、IL-8和MCP-1，内毒素(100 ng/ml)可进一步促进其分泌。共培养细胞协同分泌IL-6和IL-8部分是通过细胞与细胞的直接接触和由肿瘤坏死因子介导的。共培养的条件培养液可刺激PHM1-41心肌细胞的收缩，孕酮可抑制这种作用。孕酮和IL-10均抑制脂多糖刺激的IL-6和IL-8的分泌，孕酮也抑制趋化因子的分泌。这些数据表明，单核细胞在分娩时渗入子宫肌层，参与了促进促炎细胞因子分泌的串扰，这种作用被脂多糖增强，并可以增强心肌细胞的收缩。这些作用都被黄体酮部分抑制。
Both term and preterm parturition are characterized by an influx of macrophages and neutrophils into the myometrium and cervix, with co-incident increased peripheral blood monocyte activation. Infection and inflammation are strongly implicated in the pathology of preterm labour (PTL), with progesterone considered a promising candidate for its prevention or treatment. In this study, we investigated the effect of monocytes on myometrial smooth muscle cell inflammatory cytokine production both alone and in response to LPS, a TLR4 agonist used to trigger PTL in vivo. We also investigated the effect of monocytes on myocyte contraction. Monocytes, isolated from peripheral blood samples from term pregnant women, were cultured alone, or co-cultured with PHM1-41 myometrial smooth muscle cells, for 24 h. In a third set of experiments, PHM1-41 myocytes were cultured for 24 h in isolation. Cytokine secretion was determined by ELISA or multiplex assays. Co-culture of monocytes and myocytes led to synergistic secretion of pro-inflammatory cytokines and chemokines including IL-6, IL-8 and MCP-1, with the secretion being further enhanced by LPS (100 ng/ml). The synergistic secretion of IL-6 and IL-8 from co-cultures was mediated in part by direct cell–cell contact, and by TNF. Conditioned media from co-cultures stimulated contraction of PHM1-41 myocytes, and the effect was inhibited by progesterone. Both progesterone and IL-10 inhibited LPS-stimulated IL-6 and IL-8 secretion from co-cultures, while progesterone also inhibited chemokine secretion. These data suggest that monocytes infiltrating the myometrium at labour participate in crosstalk that potentiates pro-inflammatory cytokine secretion, an effect that is enhanced by LPS, and can augment myocyte contraction. These effects are all partially inhibited by progesterone.