Department of Infectious Diseases, The First Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
Department of Infectious Diseases, The Central Hospital of Baoji, Baoji, Shannxi 721008, P.R. China.
Int J Mol Med. 2018 May;41(5):2545-2552. doi: 10.3892/ijmm.2018.3500. Epub 2018 Feb 16.
The effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on the proliferation of hepatic stellate cells (HSCs) is largely unknown. The purpose of this study was to explore the mechanism of action of hUC‑MSCs on the proliferation of HSCs in vitro. The upper and lower double-cell co-culture system was established between hUC‑MSCs and HSCs in the experimental group. HSCs were cultured alone as a negative control group. Cell proliferation and apoptosis were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively. Cell supernatants were harvested to determine the concentration of transforming growth factor-β1 (TGF-β1) by ELISA. mRNA and protein of TGF-β1, Smad3 and Smad7 in HSCs were determined by reverse transcription-polymerase chain reaction and western blotting, respectively. In the co-culture group, the proliferation of HSCs was significantly inhibited compared with the negative control group at 24 and 48 h (p<0.05). Apoptosis of HSCs in the co-culture group increased compared with that in the negative control group, which was more obvious at 48 h (p<0.05). The concentration of TGF-β1 in the co-culture group was significantly lower than in the HSCs cultured alone (p<0.05). After HSCs were co-cultured with hUC‑MSCs for 48 h, expression of TGF-β1 and Smad3 mRNA and protein was reduced and expression of Smad7 mRNA and protein was increased compared with the negative control group (p<0.05). hUC‑MSCs inhibited proliferation of HSCs, possibly through inhibiting TGF-β1 and Smad3 expression and increasing Smad7 protein expression.
人脐带间充质干细胞(hUC-MSCs)对肝星状细胞(HSCs)增殖的影响在很大程度上尚不清楚。本研究旨在探讨 hUC-MSCs 体外作用于 HSCs 增殖的机制。实验组建立了 hUC-MSCs 与 HSCs 上下双层共培养体系,阴性对照组单独培养 HSCs。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)比色法和流式细胞术分别检测细胞增殖和凋亡情况,酶联免疫吸附试验(ELISA)检测细胞上清液中转化生长因子-β1(TGF-β1)浓度。采用逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹法(Western blot)分别检测 HSCs 中 TGF-β1、Smad3 和 Smad7 的 mRNA 和蛋白表达。与阴性对照组相比,共培养组 24 和 48 h 时 HSCs 增殖明显受到抑制(p<0.05)。与阴性对照组相比,共培养组 HSCs 凋亡增加,48 h 时更为明显(p<0.05)。与单独培养的 HSCs 相比,共培养组 TGF-β1 浓度明显降低(p<0.05)。与阴性对照组相比,hUC-MSCs 与 HSCs 共培养 48 h 后,TGF-β1 和 Smad3 mRNA 及蛋白表达降低,Smad7 mRNA 及蛋白表达升高(p<0.05)。hUC-MSCs 抑制 HSCs 增殖,可能通过抑制 TGF-β1 和 Smad3 表达,增加 Smad7 蛋白表达。