Jamhiri Iman, Hosseini Seyed Younes, Mehrabani Davood, Khodabandeh Zahra, Yaghobi Ramin, Dowran Razieh, Zahri Saber
Department of Biology, Cell and Molecular Laboratory, Faculty of Science, University of Mohaghegh Ardabili, Ardabil 56199-11367, Iran.
Department of Bacteriology and Virology, Shiraz University of Medical Sciences, Shiraz 71348-14336, Iran.
Biomed Rep. 2017 Aug;7(2):173-178. doi: 10.3892/br.2017.931. Epub 2017 Jun 21.
Activation of hepatic stellate cells (HSCs) is the pivotal event during liver fibrosis. Interleukin (IL)-24/melanoma differentiation-associated gene-7 (mda-7) has attracted attention in the pathophysiology of some diseases, while its role in activation/suppression of human HSCs is still unclear. It is important to elucidate whether the expression levels of the IL-24/mda-7 protein and its receptors in HSC cells are changed following activation. LX-2 cells, a human hepatic stellate cell line were activated by a combination of leptin and serum starvation. The activation state was evaluated through measuring the mRNA expression of profibrotic molecules, collagen-I, TIMP metalloproteinase inhibitor-1 and transforming growth factor-β. The expression of IL-24/mda-7 was assessed in mRNA and protein levels by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and ELISA methods, respectively. Hence, the amount of IL-22R1 and IL-20R2 subunit expression was also compared in activated and normal LX-2 cells by RT-qPCR. The expression level of IL-24/mda-7 and its cognate receptors was detectable both in the normal and activated LX-2 cell line. Furthermore, in activated LX-2, a significant increase of IL24 expression either on IL-22R1 and IL-20R2 subunits was also noticeable in comparison to normal cells. The activation state of LX-2 cells caused significant changes of IL-24/mda-7 and its receptors expression. In addition, the elevation in IL-24/mda-7 during LX-2 cell activation, suggested that IL-24/mda-7 and its cognate receptors serve a possible role in the development of the fibrosis process. Therefore, IL-24/mda-7 and relevant signaling pathways may be employed as a target for fibrosis treatment.
肝星状细胞(HSCs)的激活是肝纤维化过程中的关键事件。白细胞介素(IL)-24/黑色素瘤分化相关基因-7(mda-7)在某些疾病的病理生理学中受到关注,但其在人HSCs激活/抑制中的作用仍不清楚。阐明HSC细胞中IL-24/mda-7蛋白及其受体的表达水平在激活后是否发生变化很重要。人肝星状细胞系LX-2细胞通过瘦素和血清饥饿联合激活。通过测量促纤维化分子、I型胶原、TIMP金属蛋白酶抑制剂-1和转化生长因子-β的mRNA表达来评估激活状态。分别通过逆转录定量聚合酶链反应(RT-qPCR)和ELISA方法在mRNA和蛋白水平评估IL-24/mda-7的表达。因此,还通过RT-qPCR比较了激活的和正常的LX-2细胞中IL-22R1和IL-20R2亚基表达量。在正常和激活的LX-2细胞系中均能检测到IL-24/mda-7及其同源受体的表达水平。此外,与正常细胞相比,在激活的LX-2细胞中,IL-22R1和IL-20R2亚基上IL-24的表达也显著增加。LX-2细胞的激活状态导致IL-24/mda-7及其受体表达发生显著变化。此外,LX-2细胞激活过程中IL-24/mda-7的升高表明,IL-24/mda-7及其同源受体可能在纤维化过程的发展中起作用。因此,IL-24/mda-7及相关信号通路可能被用作纤维化治疗的靶点。