PMID- 32459063 OWN - NLM STAT- MEDLINE DCOM- 20211026 LR - 20220325 IS - 1755-5949 (Electronic) IS - 1755-5930 (Print) IS - 1755-5930 (Linking) VI - 26 IP - 9 DP - 2020 Sep TI - VX-765 enhances autophagy of human umbilical cord mesenchymal stem cells against stroke-induced apoptosis and inflammatory responses via AMPK/mTOR signaling pathway. PG - 952-961 LID - 10.1111/cns.13400 [doi] AB - INTRODUCTION: To investigate the protective effect of VX-765 on human umbilical mesenchymal stem cells (HUMSCs) in stroke and its mechanism. MATERIALS AND METHODS: Mouse models of ischemic stroke were established using the distal middle cerebral artery occlusion (dMCAO) method. The dMCAO mice were accordingly transplanted with HUMSCs, VX-765-treated HUMSCs, or VX-765 + MHY185-treated HUMSCs. The HUMSCs were inserted with green fluorescent protein (GFP) for measurement of transplantation efficiency which was determined by immunofluorescence assay. Oxygen-glucose deprivation (OGD) was applied to mimic ischemic environment in vitro experiments, and the HUMSCs herein were transfected with AMPK inhibitor Compound C or autophagy inhibitor 3-MA. MTT assay was used to test the toxicity of VX-765. TUNEL staining and ELISA were applied to measure the levels of apoptosis and inflammatory cytokines (IL-1beta, IL-6, and IL-10), respectively. The expressions of autophagy-associated proteins, AMPK, and mTOR were detected by Western blotting. TTC staining was applied to reveal the infarct lesions in the brain of dMCAO mice. RESULTS: The pro-inflammatory cytokines, TUNEL-positive cells, and p-mTOR were decreased while the anti-inflammatory cytokine, autophagy-related proteins, and p-AMPK were increased in HUMSCs treated with VX-765 under OGD condition. Different expression patterns were found with the above factors after transfection of 3-MA or Compound C. The pro-inflammatory cytokines, TUNEL-positive cells, and infarct sections were decreased while the anti-inflammatory cytokine and autophagy-related proteins were increased in dMCAO mice transplanted with VX-765-treated HUMSCs compared to those transplanted with HUMSCs only. The autophagy was inhibited while p-mTOR was up-regulated after transfection of MHY. CONCLUSION: VX-765 protects HUMSCs against stroke-induced apoptosis and inflammatory responses by activating autophagy via the AMPK/mTOR signaling pathway in vivo and in vitro. CI - (c) 2020 The Authors. CNS Neuroscience & Therapeutics published by John Wiley & Sons Ltd. FAU - Sun, Zhezhe AU - Sun Z AD - Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. FAU - Gu, Lei AU - Gu L AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. AD - Department of Neurology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. FAU - Wu, Ke AU - Wu K AD - Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. FAU - Wang, Kankai AU - Wang K AD - Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. FAU - Ru, Junnan AU - Ru J AD - Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. FAU - Yang, Su AU - Yang S AD - Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. FAU - Wang, Zhenzhong AU - Wang Z AD - Department of Neurosurgery, Yuyao people's Hospital, Ningbo, China. FAU - Zhuge, Qichuan AU - Zhuge Q AD - Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. FAU - Huang, Lijie AU - Huang L AD - Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. FAU - Huang, Shengwei AU - Huang S AUID- ORCID: 0000-0001-8235-6868 AD - Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. AD - Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, Wenzhou Medical University, Wenzhou, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20200527 PL - England TA - CNS Neurosci Ther JT - CNS neuroscience & therapeutics JID - 101473265 RN - 0 (Dipeptides) RN - 0 (Inflammation Mediators) RN - 0 (para-Aminobenzoates) RN - 00OLE78529 (belnacasan) RN - EC 2.7.- (Protein Kinases) RN - EC 2.7.1.1 (mTOR protein, rat) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - EC 2.7.11.3 (AMP-Activated Protein Kinase Kinases) SB - IM MH - AMP-Activated Protein Kinase Kinases MH - Animals MH - Apoptosis/drug effects/physiology MH - Autophagy/drug effects/physiology MH - Dipeptides/*pharmacology/therapeutic use MH - Humans MH - Inflammation Mediators/antagonists & inhibitors/metabolism MH - Mesenchymal Stem Cells/drug effects/*metabolism/pathology MH - Protein Kinases/*metabolism MH - Rats MH - Rats, Sprague-Dawley MH - Signal Transduction/drug effects/physiology MH - Stroke/*metabolism/pathology MH - TOR Serine-Threonine Kinases/*metabolism MH - Umbilical Cord/drug effects/*metabolism/pathology MH - para-Aminobenzoates/*pharmacology/therapeutic use PMC - PMC7415204 OTO - NOTNLM OT - VX-765 OT - autophagy OT - human umbilical mesenchymal stem cells OT - stroke COIS- The authors have no potential conflicts of interest. EDAT- 2020/05/28 06:00 MHDA- 2021/10/27 06:00 PMCR- 2020/05/27 CRDT- 2020/05/28 06:00 PHST- 2020/01/13 00:00 [received] PHST- 2020/04/09 00:00 [revised] PHST- 2020/04/28 00:00 [accepted] PHST- 2020/05/28 06:00 [pubmed] PHST- 2021/10/27 06:00 [medline] PHST- 2020/05/28 06:00 [entrez] PHST- 2020/05/27 00:00 [pmc-release] AID - CNS13400 [pii] AID - 10.1111/cns.13400 [doi] PST - ppublish SO - CNS Neurosci Ther. 2020 Sep;26(9):952-961. doi: 10.1111/cns.13400. Epub 2020 May 27.