Abdanipour Alireza, Shahsavandi Behnaz, Alipour Mohsen, Feizi Hadi
Department of Anatomical Sciences, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Department of Physiology and Pharmacology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Cell J. 2018 Jul;20(2):183-187. doi: 10.22074/cellj.2018.5164. Epub 2018 Mar 18.
Ghrelin is a peptide which has a proliferative and antiapoptotic effect in many cells including bone marrow stromal cells (BMSCs). Homeobox protein B4 (HOXB4) is a transcription factor involved in stem cell regeneration and survival. The aim of the study was to find out the efect of ghrelin on Hoxb4 expression in BMSCs.
In this experimental study, rat BMSCs were cultivated in Dulbecco's Modified Eagle Medium (DMEM). Passage three BMSCs were treated with ghrelin 100 μM for 48 hours. Real-time polymerase chain reaction (PCR) was carried out from the untreated BMSCs (B), BMSCs treated with 125 μM HO (BH), BMSCs treated with 100 μM ghrelin then 125 μM HO (BGH) and BMSCs treated with 100 μM ghrelin (BG) groups. For immunofluorescence, cells were incubated with an anti-HOXB4 monoclonal antibody. Primary antibodies were visualized using the Fluorescein isothiocyanate (FITC) method. All data are presented as mean ± SEM and P<0.05 was considered as statistical significant.
Hoxb4 expression significantly increased in the BG compared with BH and BGH groups. Furthermore, 100 μM ghrelin, increased the mean of HOXB4 positive immunoreactive cells compared to the BH group.
Ghrelin probably enhances proliferation and viability of BMSCs through Hoxb4 upregulation. However, the signaling pathway and other biological outcomes of this effect should be elucidated in different stem cells.
胃饥饿素是一种在包括骨髓间充质干细胞(BMSC)在内的许多细胞中具有增殖和抗凋亡作用的肽。同源盒蛋白B4(HOXB4)是一种参与干细胞再生和存活的转录因子。本研究的目的是探究胃饥饿素对BMSC中Hoxb4表达的影响。
在本实验研究中,大鼠BMSC在杜氏改良 Eagle 培养基(DMEM)中培养。第三代BMSC用100μM胃饥饿素处理48小时。从未处理的BMSC(B)、用125μM HO处理的BMSC(BH)、先用100μM胃饥饿素然后用125μM HO处理的BMSC(BGH)以及用100μM胃饥饿素处理的BMSC(BG)组中进行实时聚合酶链反应(PCR)。对于免疫荧光,细胞与抗HOXB4单克隆抗体孵育。使用异硫氰酸荧光素(FITC)方法观察一抗。所有数据均以平均值±标准误表示,P<0.05被认为具有统计学意义。
与BH和BGH组相比,BG组中Hoxb4表达显著增加。此外,与BH组相比,100μM胃饥饿素增加了HOXB4阳性免疫反应细胞的平均值。
胃饥饿素可能通过上调Hoxb4增强BMSC的增殖和活力。然而,这种作用的信号通路和其他生物学结果应在不同的干细胞中阐明。