López-Rincón Gonzalo, Mancilla Raúl, Pereira-Suárez Ana L, Martínez-Neri Priscila A, Ochoa-Zarzosa Alejandra, Muñoz-Valle José Francisco, Estrada-Chávez Ciro
Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, A.C. (CIATEJ), Guadalajara, Mexico; Departamento de Microbiología e Inmunología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico; Departamento de Investigación y Desarrollo, Laboratorios Virbac América Latina, Guadalajara, Jalisco, Mexico.
Instituto de Investigaciones Biomédicas (IIB), UNAM, Mexico City, Mexico.
Exp Mol Pathol. 2015 Jun;98(3):517-26. doi: 10.1016/j.yexmp.2015.03.016. Epub 2015 Mar 20.
Increased levels of prolactin (PRL) have recently been associated with carcinogenesis and the exacerbation of autoimmune diseases, and might be involved in the progression of tuberculosis (TB). To investigate the relationship between PRL and prolactin receptor (PRLr) expression with inflammatory response and apoptosis in monocytes, we used THP-1 cells stimulated with antigens of the Mycobacterium bovis AN5 strain culture filtrate protein (CFP-M. bovis). Western blot (WB), real-time Polymerase chain reaction (PCR), and immunocytochemistry were performed to identify both PRL and PRLr molecules. PRL bioactivity and proinflammatory cytokine detection were assessed. The results showed that PRL and PRLr messenger RNA (mRNA) were synthesized in THP-1 monocytes induced with CFP-M. bovis at peaks of 176- and 404-fold, respectively. PRL forms of 60 and 80kDa and PRLr isoforms of 40, 50, and 65kDa were also identified as time-dependent, while 60-kDa PRL, as well as 40-, and 50-kDa PRLr, were found as soluble forms in culture media and later in the nucleus of THP-1 monocytes. PRL of 60kDa released by monocytes exhibited bioactivity in Nb2 cells, and both synthesized PRL and synthesized PRLr were related with nitrite and proinflammatory cytokine levels proapoptotic activity in CFP-M. bovis-induced monocytes. Our results suggest the overexpression of a full-autocrine loop of PRL and PRLr in monocytes that enhances the inflammatory response and apoptosis after priming with M. bovis antigens.
催乳素（PRL）水平升高最近被认为与致癌作用以及自身免疫性疾病的加重有关，并且可能参与结核病（TB）的进展。为了研究PRL和催乳素受体（PRLr）表达与单核细胞炎症反应和凋亡之间的关系，我们使用了经牛分枝杆菌AN5株培养滤液蛋白（CFP - M. bovis）抗原刺激的THP - 1细胞。进行蛋白质免疫印迹（WB）、实时聚合酶链反应（PCR）和免疫细胞化学以鉴定PRL和PRLr分子。评估了PRL生物活性和促炎细胞因子检测。结果表明，在CFP - M. bovis诱导的THP - 1单核细胞中分别合成了PRL和PRLr信使核糖核酸（mRNA），峰值分别为176倍和404倍。还鉴定出60kDa和80kDa的PRL形式以及40kDa、50kDa和65kDa的PRLr异构体呈时间依赖性，而60kDa的PRL以及40kDa和50kDa的PRLr在培养基中以及后来在THP - 1单核细胞核中以可溶性形式存在。单核细胞释放的60kDa PRL在Nb2细胞中表现出生物活性，并且合成的PRL和合成的PRLr均与CFP - M. bovis诱导的单核细胞中的亚硝酸盐和促炎细胞因子水平及促凋亡活性有关。我们的结果表明，单核细胞中PRL和PRLr的完全自分泌环过表达，在用牛分枝杆菌抗原启动后增强了炎症反应和凋亡。
