Synergistic Protection of N-Acetylcysteine and Ascorbic Acid 2-Phosphate on Human Mesenchymal Stem cells Against Mitoptosis, Necroptosis and Apoptosis

Abstract

Human mesenchymal stem cells (hMSCs) contribute to ischemic tissue repair, regeneration, and possess ability to self-renew. However, poor viability of transplanted hMSCs within ischemic tissues has limited its therapeutic efficiency. Therefore, it is urgent to explore new method to improve the viability of the grafted cells. By using a systematic analysis, we reveal the mechanism of synergistic protection of N-acetylcysteine (NAC) and ascorbic acid 2-phosphate (AAP) on hMSCs that were under H2O2-induced oxidative stress. The combined treatment of NAC and AAP (NAC/AAP) reduces reactive oxygen species (ROS) generation, stabilizes mitochondrial membrane potential and decreases mitochondrial fission/fragmentation due to oxidative stress. Mitochondrial fission/fragmentation is a major prologue of mitoptosis. NAC/AAP prevents apoptotic cell death via decreasing the activation of BAX, increasing the expression of BCL2, and reducing cytochrome c release from mitochondria that might lead to the activation of caspase cascade. Stabilization of mitochondria also prevents the release of AIF, and its nuclear translocation which may activate necroptosis via H2AX pathway. The decreasing of mitoptosis is further studied by MicroP image analysis, and is associated with decreased activation of Drp1. In conclusion, NAC/AAP protects mitochondria from H2O2-induced oxidative stress and rescues hMSCs from mitoptosis, necroptosis and apoptosis.

Hydrogen peroxide induces programmed necrosis in rat nucleus pulposus cells through the RIP1/RIP3‐PARP‐AIF pathway

“…In our study, we found that inhibiting necroptosis evidently reduced intracellular ROS levels and raised ΔΨm levels, which indicated that the PARP‐AIF pathway might be a downstream component of RIP1/RIP3‐mediated programmed necrosis in H 2 O 2 ‐induced NP cells death. In addition, oxidative stress has been shown to induce cell death in human bone mesenchymal stem cells (BMSC) and hepatocellular carcinoma cells through PARP hyperactivation‐mediated necroptosis . However, there have been no reports indicating whether the PARP‐AIF pathway is involved in H 2 O 2 ‐mediated NP cells death.…”

“…In addition, oxidative stress has been shown to induce cell death in human bone mesenchymal stem cells (BMSC) and hepatocellular carcinoma cells through PARP hyperactivation-mediated necroptosis. 18,50 However, there have been no reports indicating whether the PARP-AIF pathway is involved in H 2 O 2 -mediated NP cells death.…”

“…Subsequently, AIF is released from the depolarized mitochondria, translocates to the nucleus, and then binds with DNA to promote caspase‐independent chromatinolysis . Necroptosis induced by oxidative stress requires RIP1/RIP3 activation and PARP hyperactivation . However, the interaction mechanism between RIP1/RIP3 and PARP‐AIF is still ambiguous.…”

“…16,17 Necroptosis induced by oxidative stress requires RIP1/RIP3 activation and PARP hyperactivation. 18,19 However, the interaction mechanism between RIP1/ RIP3 and PARP-AIF is still ambiguous.…”

Enhancement of Mitochondrial Transfer by Antioxidants in Human Mesenchymal Stem Cells

“…In addition, MCA also helped hMSCs to cope with H 2 O 2 -induced cell death by inhibiting cell death and maintaining mitochondria integrity [16]. …”