Interleukin-33 regulates the endoplasmic reticulum stress of human myometrium via an influx of calcium during initiation of labor.
Interleukin-33 regulates the endoplasmic reticulum stress of human myometrium via an influx of calcium during initiation of labor.
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DOI：
10.7554/elife.75072
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发表时间：
2022-08-23
期刊：
影响因子：
7.7
通讯作者：
Zhang, Guoying
中科院分区：
文献类型：
--
作者：
Chen, Li;Song, Zhenzhen;Cao, Xiaowan;Fan, Mingsong;Zhou, Yan;Zhang, Guoying
目前，炎症被认为是早产的主要原因之一。作为白细胞介素1β（IL-1β）家族的一员，白细胞介素33（IL-33）已被证明参与正常妊娠以及多种妊娠相关疾病。本研究旨在探讨 IL-33 在分娩过程中子宫平滑肌细胞中的潜在功能。足月妊娠（妊娠≥37周）女性的子宫肌层样本要么被冷冻，要么被分离和培养。使用免疫组织化学和蛋白质印迹来评估 IL-33 的分布。将培养的细胞与脂多糖 (LPS) 一起孵育以模拟炎症，并在 4μ8C（IRE1 抑制剂 III）存在下孵育以阻断内质网 (ER) 应激，以及 BAPTA-AM（一种钙螯合剂）。 LPS限时降低核IL-33的表达并诱导ER应激。然而，敲低 IL-33 会增加 LPS 诱导的钙浓度、内质网应激以及核因子 kappa-B (NF-κB) 和 P38 丝裂原激活蛋白激酶 (P38 MAPK) 的磷酸化。此外，siRNA IL-33 通过 NF-κB 和 p38 途径进一步刺激 LPS 增强的环氧合酶-2 (COX-2) 表达。随着临产的开始，细胞核中 IL-33 的表达减少。 LPS 诱导 ER 应激，并增加培养的子宫肌细胞中分娩相关基因 COX-2 以及 IL-6 和 IL-8 的表达。 IL-33也增加了COX-2的表达，但在其被敲低后，LPS对钙的刺激作用增强。 4μ8C也显着抑制COX-2的表达。膜上钙通道和细胞内游离钙离子的表达均增加，并伴有磷酸化的NF-κB和p38。这些数据表明，IL-33 可能通过钙离子流入人子宫平滑肌细胞而导致内质网应激，从而参与分娩的启动。该研究得到国家自然科学基金项目（No. 81300507）的资助。
Inflammation is currently recognized as one of the major causes of premature delivery. As a member of the interleukin-1β (IL-1β) family, interleukin-33 (IL-33) has been shown to be involved in normal pregnancy as well as a variety of pregnancy-related disorder. This study aims to investigate the potential function of IL-33 in uterine smooth muscle cells during labor. Myometrium samples from term pregnant (≥37 weeks gestation) women were either frozen or cells were isolated and cultured. Immunohistochemistry and western blotting were used to assess the distribution of IL-33. Cultured cells were incubated with lipopolysaccharide (LPS) to mimic inflammation as well as in the presence of 4μ8C (IRE1 inhibitor III) to block endoplasmic reticulum (ER) stress and BAPTA-AM, a calcium chelator. LPS reduced the expression of nuclear IL-33 in a time-limited manner and induced ER stress. However, knockdown of IL-33 increased LPS-induced calcium concentration, ER stress and phosphorylation of nuclear factor kappa-B (NF-κB), and P38 mitogen-activated protein kinase (P38 MAPK). In addition, siRNA IL-33 further stimulates LPS enhanced cyclooxygenase-2 (COX-2) expression via NF-κB and p38 pathways. IL-33 expression was decreased in the nucleus with the onset of labor. LPS-induced ER stress and increased expression of the labor-associated gene, COX-2, as well as IL-6 and IL-8 in cultured myometrial cells. IL-33 also increased COX-2 expression, but after it was knocked down, the stimulating effect of LPS on calcium was enhanced. 4μ8C also inhibited the expression of COX-2 markedly. The expression of calcium channels on the membrane and intracellular free calcium ion were both increased which was accompanied by phosphorylated NF-κB and p38. These data suggest that IL-33 may be involved in the initiation of labor by leading to stress of the ER via an influx of calcium ions in human uterine smooth muscle cells. This study was supported by grants from the National Natural Science Foundation of China (No. 81300507).