Department of Gastroenterology, Shenyang General Hospital of PLA, No. 83 Wenhua Road Shenyang City, 110016, Liaoning, PR China.
Department of Gastroenterology, Shenyang General Hospital of PLA, No. 83 Wenhua Road Shenyang City, 110016, Liaoning, PR China.
Chem Biol Interact. 2019 Jan 5;297:109-118. doi: 10.1016/j.cbi.2018.10.019. Epub 2018 Oct 23.
β2m-/Thy1+ bone marrow-derived hepatocyte stem cells (BDHSCs) have a potential to be applied for cellular treatment in liver cirrhosis. However, the resultant tissue regeneration is restricted by transplanted cells' death. The accumulation of transforming growth factor beta 1 (TGF-β1) in liver fibrosis local microenvironment may play an essential role in the rapid cell death of implanted β2m-/Thy1+ BDHSCs. The main mechanism of poor survival of the target stem cells is still unknown. Delphinidin, an anthocyanidin, has potent antioxidant and anti-inflammatory activities. However, whether this bio-active ingredient can substantially contribute to β2m-/Thy1+ BDHSCs' protection from TGF-β1 induced apoptosis in vitro remains to be elucidated. In the present research, we determined whether delphinidin pretreatment can improve the survival of β2m-/Thy1+ BDHSCs during exposure to TGF-β1 and elucidated its underlying mechanisms. By using TGF-β1, we induced the apoptosis of β2m-/Thy1+ BDHSCs and assessed the apoptotic rates up to 24 h by flow cytometry. β2m-/Thy1+ BDHSC proliferation was gauged using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl- 2H-tetrazolium bromide (MTT) assay. The expression grades of Bcl-2, Akt, caspase-3, and Bax were observed through Western blot analysis. We found that delphinidin can significantly impede TGF-β1-induced apoptosis dose-dependently, scavenge reactive oxygen species (ROS), and inhibit the discharge of caspase-3 in β2m-/Thy1+ BDHSCs. We also demonstrated that delphinidin can activate the phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathway. The suppression of ROS and succeeding apoptosis was achieved by pretreatment with LY294002, a PI3K/Akt pathway inhibitor. In summary, our findings revealed that delphinidin can protect β2m-/Thy1+ BDHSCs from apoptosis and ROS-dependent oxidative stress induced by the TGF-β1 via PI3K/Akt signaling pathway. On the basis of these data, delphinidin can be regarded as a promising anti-apoptotic agent for enhancing β2m-/Thy1+ BDHSC survival during cell transplantation in liver cirrhosis patients.
β2m-/Thy1+ 骨髓源性肝干细胞（BDHSCs）具有作为细胞疗法应用于肝硬化的潜力。然而，由于移植细胞的死亡，导致组织再生受到限制。转化生长因子β 1（TGF-β1）在肝纤维化局部微环境中的积累可能在植入的β2m-/Thy1+BDHSCs 快速细胞死亡中发挥重要作用。靶干细胞存活率低的主要机制仍不清楚。矢车菊素是一种花色苷，具有很强的抗氧化和抗炎活性。然而，这种生物活性成分是否能显著促进β2m-/Thy1+BDHSCs 免受 TGF-β1 诱导的体外细胞凋亡，仍有待阐明。在本研究中，我们确定了矢车菊素预处理是否可以提高β2m-/Thy1+BDHSCs 在暴露于 TGF-β1 时的存活率，并阐明了其潜在机制。通过使用 TGF-β1，我们诱导β2m-/Thy1+BDHSCs 凋亡，并通过流式细胞术评估高达 24 小时的凋亡率。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐（MTT）测定评估β2m-/Thy1+BDHSC 的增殖。通过 Western blot 分析观察 Bcl-2、Akt、caspase-3 和 Bax 的表达水平。我们发现，矢车菊素可以显著抑制 TGF-β1 诱导的凋亡，呈剂量依赖性，清除活性氧（ROS），并抑制β2m-/Thy1+BDHSCs 中 caspase-3 的释放。我们还表明，矢车菊素可以激活磷脂酰肌醇-3-激酶（PI3K）/Akt 信号通路。用 PI3K/Akt 通路抑制剂 LY294002 预处理可抑制 ROS，继而抑制细胞凋亡。总之，我们的研究结果表明，矢车菊素可以通过 PI3K/Akt 信号通路保护β2m-/Thy1+BDHSCs 免受 TGF-β1 诱导的凋亡和 ROS 依赖性氧化应激。基于这些数据，矢车菊素可以被视为一种有前途的抗凋亡剂，用于增强肝硬化患者细胞移植过程中β2m-/Thy1+BDHSC 的存活率。
