Prakash Ravi, Fauzia Eram, Siddiqui Abu Junaid, Yadav Santosh Kumar, Kumari Neha, Singhai Atin, Khan Mohsin Ali, Janowski Miroslaw, Bhutia Sujit Kumar, Raza Syed Shadab
Laboratory for Stem Cell & Restorative Neurology, Department of Biotechnology, Era's Lucknow Medical College Hospital, Era University, Sarfarazganj, Lucknow, 226003, India.
Department of Pathology, King George's Medical University, Lucknow, 226020, India.
Stem Cell Rev Rep. 2021 Dec;17(6):2347-2358. doi: 10.1007/s12015-021-10212-z. Epub 2021 Sep 6.
Stem cell therapies are becoming increasingly popular solutions for neurological disorders. However, there is a lower survival rate of these cells after transplantation. Oxidative stress is linked to brain damage, and it may also impact transplanted stem cells. To better understand how transplanted cells respond to oxidative stress, the current study used HO. We briefly illustrated that exogenous HO treatment exaggerated oxidative stress in the human dental pulp and mesenchymal stem cells. 2',7'-Dichlorofluorescin diacetate (DCFDA), MitoSOX confirms the reactive oxygen species (ROS) involvement, which was remarkably subsided by the ROS inhibitors. The findings showed that HO activates autophagy by enhancing pro-autophagic proteins, Beclin1 and Atg7. Increased LC3II/I expression (which co-localized with lysosomal proteins, LAMP1 and Cathepsin B) showed that HO treatment promoted autophagolysosome formation. In the results, both Beclin1 and Atg7 were observed co-localized in mitochondria, indicating their involvement in mitophagy. The evaluation of Erk1/2 in the presence and absence of Na-Pyruvate, PEG-Catalase, and PD98059 established ROS-Erk1/2 participation in autophagy regulation. Further, these findings showed a link between apoptosis and autophagy. The results conclude that HO acts as a stressor, promoting autophagy and mitophagy in stem cells under oxidative stress. The current study may help understand better cell survival and death approaches for transplanted cells in various neurological diseases. The current study uses human Dental Pulp and Mesenchymal Stem cells to demonstrate the importance of HO-driven autophagy in deciding the fate of these cells in an oxidative microenvironment. To summarise, we discovered that exogenous HO treatment causes oxidative stress. Exogenous HO treatment also increased ROS production, especially intracellular HO. HO stimulated the ErK1/2 signaling pathway and autophagy. Erk1/2 was found to cause autophagy. Further, the function of mitophagy appeared to be an important factor in the HO-induced regulation of these two human stem cell types. In a nutshell, by engaging in autophagy nucleation, maturation, and terminal phase proteins, we elucidated the participation of autophagy in cell dysfunction and death.
干细胞疗法正日益成为治疗神经疾病的热门解决方案。然而，这些细胞在移植后的存活率较低。氧化应激与脑损伤有关，它也可能影响移植的干细胞。为了更好地了解移植细胞对氧化应激的反应，当前的研究使用了血红素加氧酶（HO）。我们简要说明了外源性HO处理会加剧人牙髓和间充质干细胞中的氧化应激。2',7'-二氯荧光素二乙酸酯（DCFDA）、MitoSOX证实了活性氧（ROS）的参与，而ROS抑制剂可显著减轻这种参与。研究结果表明，HO通过增强自噬相关蛋白Beclin1和Atg7来激活自噬。LC3II/I表达增加（与溶酶体蛋白LAMP1和组织蛋白酶B共定位）表明HO处理促进了自噬溶酶体的形成。结果显示，Beclin1和Atg7均在线粒体中共定位，表明它们参与了线粒体自噬。在有无丙酮酸钠、聚乙二醇过氧化氢酶和PD98059的情况下对Erk1/2进行评估，证实了ROS-Erk1/2参与自噬调节。此外，这些研究结果表明了细胞凋亡与自噬之间的联系。结果得出结论，HO作为一种应激源，在氧化应激下促进干细胞中的自噬和线粒体自噬。当前的研究可能有助于更好地理解各种神经疾病中移植细胞的存活和死亡方式。当前的研究使用人牙髓和间充质干细胞来证明HO驱动的自噬在决定这些细胞在氧化微环境中的命运方面的重要性。总之，我们发现外源性HO处理会导致氧化应激。外源性HO处理还会增加ROS的产生，尤其是细胞内的HO。HO刺激了ErK1/2信号通路和自噬。发现Erk1/2会导致自噬。此外，线粒体自噬的功能似乎是HO诱导调节这两种人类干细胞类型的一个重要因素。简而言之，通过参与自噬的成核、成熟和终末期蛋白，我们阐明了自噬在细胞功能障碍和死亡中的参与情况。
