Suppressing phosphatidylcholine‐specific phospholipase C and elevating ROS level, NADPH oxidase activity and Rb level induced neuronal differentiation in mesenchymal stem cells

Abstract

In the previous research, we found that D609 (tricyclodecan-9-yl-xanthogenate) could induce human marrow stromal cell (hMSC) differentiation to neuron-like cells. In this study, to understand the possible mechanism, we sequentially investigated the changes of phosphatidylcholine-specific phospholipase C (PC-PLC) activity, the expression of Rb, the intracellular reactive oxygen species (ROS) levels, NADPH oxidase and superoxide dismutase (SOD) activities when D609 induced neuronal differentiation in rat mesenchymal stem cells (MSCs). The results showed that D609 obviously inhibited the activity of PC-PLC when it induced neuronal differentiation in rat MSCs. Simultaneously, ROS level and the activity of NADPH oxidase increased significantly, but the MnSOD and Cu/ZnSOD activities were not altered. Furthermore, the level of Rb protein was evidently elevated. Our data suggested that PC-PLC mediated neuronal differentiation of rat MSCs by elevating NADPH oxidase activity, ROS level, and up-regulating the expression of Rb protein.

Extract of Curcuma longa L. and (-)-Epigallo Catechin-3-Gallate Enhanced Proliferation of Adipose Tissue–derived Mesenchymal Stem Cells (AD-MSCs) and Differentiation of AD-MSCs into Endothelial Progenitor Cells

“…An increasing number of studies reported on the crucial role of ROS/NO on MSCs differentiation [28]. The neuronal differentiation of MSCs involved upregulation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and increased ROS generation [29]. Our results supported that ECL and EGCG could enhance MSCs differentiation into EPCs.…”

“…ROS generation through the activity of the NADPH oxidase (Nox)-2 and Nox-4 isoform has been demonstrated in human CD34 + cells which may contribute to the activation of intracellular signaling pathways leading to mitochondriogenesis, cell survival, and differentiation in hematopoietic stem cells [31]. ROS as well as reactive nitrogen species (RNS) are involved in stem cell mobilization, function and differentiation in a very complex way [29].…”

Effect of inhibitors of NADPH oxidase complex and mitochondrial ATP‐sensitive potassium channels on generation of dopaminergic neurons from neurospheres of mesencephalic precursors

“…The number of proliferating cells further decreases after induction of the differentiation, as expected after withdrawal of the mitogens from the cell culture (Studer et al, 2000;Ebert et al, 2008). Several previous studies have suggested that intracellular levels of ROS modulate neuronal differentiation, and it has been observed that cell fractions exhibiting low levels of ROS contain proliferative progenitor cells, while newly generated neurons express high levels of ROS (Katoh et al, 1997;Goldsmit et al, 2001;Wang et al, 2007;Tsatmali et al, 2005); this was also confirmed in the present study by DHE labeling. Because undifferentiated neural stem cells contain very low levels of ROS, it is reasonable to believe that further ROS inhibition has no significant effects.…”

“…In previous studies with antioxidants (Rodriguez-Pallares et al, 2001 and preliminary experiments with Apo or 5-HD, we did not observe effects on DA cell generation when treatments were administered during the first week of proliferation (i.e., 1 to 7 DIV). This is in accordance with previous studies that observed that proliferative progenitor cell exhibited very low levels of ROS, while ROS increased to high levels in newly generated neurons (Katoh et al, 1997;Goldsmit et al, 2001;Tsatmali et al, 2005;Wang et al, 2007). Therefore, treatments were added to the medium during the late proliferation period (i.e., 7 to 14 DIV) or the differentiation period (i.e., 14 to 21DIV), for an incubation period of 7 days (i.e., 7 days of treatment), except when otherwise stated.…”

Stem cells and the impact of ROS signaling

“…However, the study of metabolism and ROS mediated mechanisms of stem cell fate regulation has led to improved differentiation and reprogramming protocols. Differentiation of ESCs towards the cardiac lineage has been shown to rely on H 2 O 2 signaling induced by NOX4 upregulation (Wang et al, 2007;Xiao et al, 2009). In the case of the cardiac lineage, not only are ROS important for differentiation, but the exclusive use of oxidative phosphorylation in cardiomyocytes compared with pluripotent stem cells (PSCs) can be taken advantage of to improve purification and differentiation efficiency (Chung et al, 2007;Tohyama et al, 2012).…”