Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco A,C, Guadalajara, Jalisco 44270, México.
J Inflamm (Lond). 2013 Jun 3;10(1):24. doi: 10.1186/1476-9255-10-24.
Prolactin from pituitary gland helps maintain homeostasis but it is also released in immune cells where its function is not completely understood. Pleiotropic functions of prolactin (PRL) might be mediated by different isoforms of its receptor (PRLr).
The aim of this study was to investigate the relationship between the eventual synthesis of PRL and PRLr isoforms with the inflammatory response in monocytes. We used THP-1 and monocytes isolated from healthy subjects stimulated with lipopolysaccharide (LPS). Western blot, real time PCR and immunocytochemistry were performed to identify both molecules. The bioactivity of the PRL was assessed using a bioassay and ELISA to detect pro inflammatory cytokines.
PRLr mRNA and PRL mRNA were synthesized in THP-1 monocytes activated with LPS with peaks of 300-fold and 130-fold, respectively. The long (100 kDa) and the intermediate (50 kDa) isoforms of PRLr and big PRL (60 kDa) were time-dependent upregulated for monocytes stimulated with LPS. This expression was confirmed in monocytes from healthy subjects. The PRLr intermediate isoform and the big PRL were found soluble in the culture media and later in the nucleus in THP-1 monocytes stimulated with LPS. Big PRL released by monocytes showed bioactivity in Nb2 Cells, and both PRL and PRLr, synthesized by monocytes were related with levels of nitrites and proinflammatory citokines.
Our results suggest the expression of a full-autocrine loop of PRL enhances the inflammatory response in activated monocytes. This response mediated by big PRL may contribute to the eradication of potential pathogens during innate immune response in monocytes but may also contribute to inflammatory disorders.
垂体分泌的催乳素有助于维持体内平衡，但也会在免疫细胞中释放，其功能尚不完全清楚。催乳素（PRL）的多效性功能可能由其受体（PRLr）的不同同工型介导。
本研究旨在探讨最终催乳素和 PRLr 同工型的合成与单核细胞炎症反应之间的关系。我们使用 THP-1 和从健康受试者中分离的单核细胞，用脂多糖（LPS）刺激。采用 Western blot、实时 PCR 和免疫细胞化学法鉴定这两种分子。使用生物测定法和 ELISA 检测促炎细胞因子来评估 PRL 的生物活性。
用 LPS 激活的 THP-1 单核细胞中合成了 PRLr mRNA 和 PRL mRNA，分别达到了 300 倍和 130 倍的峰值。LPS 刺激的单核细胞中，PRLr 的长（100 kDa）和中（50 kDa）同工型以及大 PRL（60 kDa）同工型呈时间依赖性上调。这一表达在来自健康受试者的单核细胞中得到了证实。在 LPS 刺激的 THP-1 单核细胞中，发现 PRLr 中间同工型和大 PRL 可溶，并随后在细胞核中表达。单核细胞释放的大 PRL 在 Nb2 细胞中具有生物活性，单核细胞合成的 PRL 和 PRLr 与亚硝酸盐和促炎细胞因子水平相关。
我们的结果表明，PRL 的完整自分泌环的表达增强了激活单核细胞的炎症反应。这种由大 PRL 介导的反应可能有助于单核细胞固有免疫反应中潜在病原体的清除，但也可能导致炎症性疾病。
Zotero 插件
