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在生理和病理条件下,极化的牛子宫内膜上皮细胞在向量上分泌前列腺素和趋化因子。

Polarised bovine endometrial epithelial cells vectorially secrete prostaglandins and chemotactic factors under physiological and pathological conditions.

机构信息

School of Medicine, College of Medicine, Institute of Life Science, Swansea University, Singleton Park, Swansea SA2 8PP, UK.

出版信息

Reproduction. 2013 Jan 8;145(1):57-72. doi: 10.1530/REP-12-0253. Print 2013 Jan.

Abstract

Epithelial cells of the endometrium secrete prostaglandins to regulate the bovine oestrous cycle and form a functional barrier to microbes. However, bacterial infection of the endometrium commonly causes infertility in dairy cattle by disrupting endometrial physiology. Epithelial cell cultures are used to study the mechanisms of physiology and pathology, but 2D cultures may not reflect the 3D complexity of the epithelium. In this study, a polarised epithelial cell transwell culture was developed, using transepithelial resistance (TER), to monitor epithelial integrity. Polarised epithelial cells were treated with oxytocin and arachidonic acid to test physiological function and with lipopolysaccharide (LPS) to mimic bacterial infection. Supernatants were analysed for prostaglandin E(2) (PGE), prostaglandin F(2)(α), the chemokine interleukin-8 (IL8) and the ability of supernatants to induce neutrophil migration. Confluent epithelial cells established polarity when TER was >1800  Ω cm(2) and predominantly released prostaglandins basolaterally. In contrast, IL8 from epithelial cells accumulated apically and the supernatants were highly chemotactic for neutrophils. The striking exception was when the epithelial cells were treated with LPS in the apical or basolateral compartment independently, which led to the release of IL8 towards the treated compartment. Although stromal cells also accumulated PGE and IL8 in response to treatment, co-culture of stromal cells in the well below polarised epithelial cells did not influence cellular responses. In conclusion, polarised endometrial epithelial cells vectorially released prostaglandins and chemokines to reflect their respective mechanistic roles in physiology and pathology.

摘要

子宫内膜上皮细胞分泌前列腺素以调节牛发情周期,并形成对微生物的功能性屏障。然而,细菌感染子宫内膜通常会通过破坏子宫内膜生理机能导致奶牛不孕。上皮细胞培养常用于研究生理和病理机制,但 2D 培养可能无法反映上皮细胞的 3D 复杂性。在这项研究中,通过跨上皮电阻(TER)开发了一种极化上皮细胞 Transwell 培养,以监测上皮完整性。用催产素和花生四烯酸处理极化上皮细胞以测试生理功能,用脂多糖(LPS)模拟细菌感染。分析上清液中的前列腺素 E2(PGE2)、前列腺素 F2α、趋化因子白细胞介素 8(IL8)以及上清液诱导中性粒细胞迁移的能力。当 TER>1800 Ω cm2 时,细胞达到融合状态,极性建立,主要从基底外侧释放前列腺素。相比之下,IL8 从上皮细胞中积累到顶端,并且上清液对中性粒细胞具有很强的趋化性。一个明显的例外是当上皮细胞分别在顶端或基底外侧用 LPS 处理时,导致 IL8 向处理部位释放。尽管基质细胞在受到刺激时也会释放 PGE 和 IL8,但在极化上皮细胞下方的培养孔中共同培养基质细胞不会影响细胞反应。总之,极化的子宫内膜上皮细胞以定向方式释放前列腺素和趋化因子,以反映它们在生理和病理机制中的各自作用。

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