Cao H-J, Wang M-D, Li S-G, Zhu L, Zheng J-H
Department of Gastrointestinal Medicine, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China.
Department of Emergency and Trauma Center, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China
Genet Mol Res. 2017 Mar 16;16(1):gmr-16-01-gmr.16019201. doi: 10.4238/gmr16019201.
We investigated the paracrine effects of bone marrow mesenchymal stem cells (BMSCs) on the proliferation, apoptosis, and alpha-actin-2 (ACTA2) expression of hepatic stellate cells (HSCs), and explored the possible mechanisms of hepatocyte growth factor (HGF). We established a co-culture system by culturing BMSCs on the upper layer and HSCs on the lower layer of a 6-well Transwell plate. Normal HSCs were cultured alone as a control. Cell apoptosis was determined by flow cytometry. We detected the expression of ACTA2 mRNA and ACTA2 protein in HSC using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting, respectively. ACTA2 in HSCs was detected by fluorescent staining, and HGF in the co-culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The apoptotic rate of HSCs in the experiment group was 2.6 times that in the control group (P < 0.05). The expression levels of ACTA2 mRNA and ACTA2 protein were significantly inhibited in HSCs compared with the control group (P < 0.05). HGF concentration in the co-culture supernatant was 0.43 ± 0.47 mM in the experimental group, which was significantly higher than in the control group (0.16 ± 0.43 mM) (P < 0.05). The paracrine effect of BMSCs, which was caused by the suppression of ACTA2 and HGF expression, induced HSC apoptosis.
我们研究了骨髓间充质干细胞（BMSCs）对肝星状细胞（HSCs）增殖、凋亡及α-肌动蛋白-2（ACTA2）表达的旁分泌作用，并探讨了肝细胞生长因子（HGF）的可能机制。我们通过在6孔Transwell板的上层培养BMSCs，下层培养HSCs建立了共培养体系。单独培养正常HSCs作为对照。采用流式细胞术检测细胞凋亡。分别使用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法检测HSC中ACTA2 mRNA和ACTA2蛋白的表达。通过荧光染色检测HSCs中的ACTA2，通过酶联免疫吸附测定（ELISA）检测共培养上清液中的HGF。实验组HSCs的凋亡率是对照组的2.6倍（P<0.05）。与对照组相比，HSCs中ACTA2 mRNA和ACTA2蛋白的表达水平显著受到抑制（P<0.05）。实验组共培养上清液中HGF浓度为0.43±0.47 mM，显著高于对照组（0.16±0.43 mM）（P<0.05）。BMSCs的旁分泌作用通过抑制ACTA2和HGF表达诱导HSC凋亡。
