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 +骨髓源性肝细胞干细胞（BDHSC）有潜力用于肝硬化的细胞治疗。但是，所得的组织再生受到移植细胞死亡的限制。肝纤维化局部微环境中转化生长因子β1（TGF-β1）的积累可能在植入的β2m-/ Thy1 + BDHSCs的快速细胞死亡中起重要作用。靶干细胞存活不良的主要机制仍是未知的。花青素Delphinidin具有有效的抗氧化和消炎作用。然而，这种生物活性成分是否可以实质性地促进β2m-/ Thy1 + BDHSCs免受TGF-β1诱导的体外细胞凋亡仍有待阐明。在目前的研究中，我们确定了翠雀素预处理是否可以提高TGF-β1暴露期间β2m-/ Thy1 + BDHSCs的存活率，并阐明了其潜在机制。通过使用TGF-β1，我们诱导了β2m-/ Thy1 + BDHSCs的凋亡，并通过流式细胞术评估了长达24小时的凋亡率。使用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基-2H-四唑溴化物（MTT）测定了β2m-/ Thy1 + BDHSC的增殖。通过蛋白质印迹分析观察到Bcl-2，Akt，caspase-3和Bax的表达等级。我们发现飞燕草碱可以显着地剂量依赖性地抑制TGF-β1诱导的细胞凋亡，清除活性氧（ROS），并抑制caspase-3在β2m-/ Thy1 + BDHSCs中的释放。我们还证明了delphinidin可以激活磷脂酰肌醇-3-激酶（PI3K）/ Akt信号通路。通过用PI3K / Akt途径抑制剂LY294002进行预处理，可以实现对ROS的抑制和随后的凋亡。总而言之，我们的发现表明，飞燕草碱可以保护β2m-/ Thy1 + BDHSCs免受TGF-β1通过PI3K / Akt信号通路诱导的凋亡和ROS依赖性氧化应激。根据这些数据，可以将delphinidin视为在肝硬化患者细胞移植过程中增强β2m-/ Thy1 + BDHSC存活的有前途的抗凋亡药物。