Journal of Molecular Neuroscience

, Volume 67, Issue 3, pp 400–410 | Cite as

Protective Role of SOCS3 Modified Bone Marrow Mesenchymal Stem Cells in Hypoxia-Induced Injury of PC12 Cells

  • Bin ZhouEmail author
  • Hong-Yun Liu
  • Bao-Lian Zhu


We attempted to explore the possible effects of SOCS3 (suppressor of cytokine signaling 3)-modified bone marrow mesenchymal stem cells (BMSCs) on the hypoxic injury of rat adrenal gland pheochromocytoma (PC-12) cells. PC12 cells were cultured with EGFP (enhanced green fluorescent protein)-BMSCs and SOCS3-BMSCs respectively under hypoxia in vitro and classified into control, hypoxia, EGFP-BMSCs, and SOCS3-BMSC groups. CCK-8, Hoechst 33258 staining, and Annexin V-FITC/PI staining were assessed to measure the viability and apoptosis of hypoxia-induced PC12 cells. The JAK/STAT3 pathway- and apoptosis-related proteins were identified by Western blot. Finally, rat models of permanent middle cerebral artery occlusion (pMCAO) were established to verify the potential influences of SOCS3-BMSCs in vivo. SOCS3-modified BMSCs can stably express SOCS3 protein. EGFP-BMSCs, especially SOCS3-BMSCs, can improve cell viability and SOD content, and reduce cell apoptosis, LDH viability, and MDA content in hypoxia-induced PC12 cells (all P < 0.05). Besides, EGFP-BMSCs and SOCS3-BMSCs decreased cleaved caspase-3 level and increased Bcl-2/Bax of hypoxia-induced PC12 cells, while SOCS3-BMSCs could also elevate SOCS3 protein and reduce p-STAT3 protein level in hypoxia-induced PC12 cells (all P < 0.05). In vivo experiments confirmed that EGFP-BMSCs, particularly SOCS3-BMSCs, could ameliorate infarct size and inhibit neuronal apoptosis of different degrees in pMACO rats (all P < 0.05). SOCS3-modified BMSCs can alleviate oxidative stress, improve cell viability, and reduce neuronal apoptosis by downregulation of JAK/STAT3 pathway, thereby exerting the neuroprotective role in ischemic brain injury.


SOCS3 BMSCs JAK/STAT3 Hypoxia PC12 cells 



We thank the reviewers for their insightful suggestions and comments.

Compliance with Ethical Standards

This study has got the approval from the Ethics Committee for Animal Experiments in Linyi Central Hospital, which was in strictly obedience to the Guide for the Care and Use of Laboratory Animals published by the National Institute of Health in the USA (Health 1985).

Conflicts of Interest

The authors declare that they have no conflict of interest.


  1. Baker BJ, Akhtar LN, Benveniste EN (2009) SOCS1 and SOCS3 in the control of CNS immunity. Trends Immunol 30:392–400. CrossRefGoogle Scholar
  2. Bates S, Read SJ, Harrison DC, Topp S, Morrow R, Gale D, Murdock P, Barone FC, Parsons AA, Gloger IS (2001) Characterisation of gene expression changes following permanent MCAO in the rat using subtractive hybridisation. Brain Res Mol Brain Res 93:70–80CrossRefGoogle Scholar
  3. Boyle K, Zhang JG, Nicholson SE, Trounson E, Babon JJ, McManus EJ, Nicola NA, Robb L (2009) Deletion of the SOCS box of suppressor of cytokine signaling 3 (SOCS3) in embryonic stem cells reveals SOCS box-dependent regulation of JAK but not STAT phosphorylation. Cell Signal 21:394–404. CrossRefGoogle Scholar
  4. Chen J, Li Y, Katakowski M, Chen X, Wang L, Lu D, Lu M, Gautam SC, Chopp M (2003) Intravenous bone marrow stromal cell therapy reduces apoptosis and promotes endogenous cell proliferation after stroke in female rat. J Neurosci Res 73:778–786. CrossRefGoogle Scholar
  5. Chen S, Tang Y, Qian Y, Chen R, Zhang L, Wo L, Chai H (2014) Allicin prevents H(2)O(2)-induced apoptosis of HUVECs by inhibiting an oxidative stress pathway. BMC Complement Altern Med 14:321. CrossRefGoogle Scholar
  6. Choi JS, Shin YJ, Cha JH, Kim HY, Choi JY, Chun MH, Lee MY (2008) Induction of suppressor of cytokine signaling-3 in astrocytes of the rat hippocampus following transient forebrain ischemia. Neurosci Lett 441:323–327. CrossRefGoogle Scholar
  7. Collins AS, McCoy CE, Lloyd AT, O'Farrelly C, Stevenson NJ (2013) miR-19a: an effective regulator of SOCS3 and enhancer of JAK-STAT signalling. PLoS One 8:e69090. CrossRefGoogle Scholar
  8. Crigler L, Robey RC, Asawachaicharn A, Gaupp D, Phinney DG (2006) Human mesenchymal stem cell subpopulations express a variety of neuro-regulatory molecules and promote neuronal cell survival and neuritogenesis. Exp Neurol 198:54–64. CrossRefGoogle Scholar
  9. Deng YH, Kuang SJ, Hei MY, Tian L (2006) Effects of inosine on neuronal apoptosis and the expression of cytochrome C mRNA following hypoxic-ischemic brain damage in neonatal rats. Zhongguo Dang Dai Er Ke Za Zhi 8:266–271Google Scholar
  10. Deng YB, Ye WB, Hu ZZ, Yan Y, Wang Y, Takon BF, Zhou GQ, Zhou YF (2010) Intravenously administered BMSCs reduce neuronal apoptosis and promote neuronal proliferation through the release of VEGF after stroke in rats. Neurol Res 32:148–156. CrossRefGoogle Scholar
  11. Dominguez E, Mauborgne A, Mallet J, Desclaux M, Pohl M (2010) SOCS3-mediated blockade of JAK/STAT3 signaling pathway reveals its major contribution to spinal cord neuroinflammation and mechanical allodynia after peripheral nerve injury. J Neurosci 30:5754–5766. CrossRefGoogle Scholar
  12. Gao G, Fan H, Zhang X, Zhang F, Wu H, Qi F, Zhao L, Li Y (2017) Neuroprotective effect of G(14)-humanin on global cerebral ischemia/reperfusion by activation of SOCS3 - STAT3 - MCL-1 signal transduction pathway in rats. Neurol Res 39:895–903. CrossRefGoogle Scholar
  13. Gu Q, Kong Y, Yu ZB, Bai L, Xiao YB (2011) Hypoxia-induced SOCS3 is limiting STAT3 phosphorylation and NF-kappaB activation in congenital heart disease. Biochimie 93:909–920. CrossRefGoogle Scholar
  14. Health USNIo (1985) Laboratory animal welfare: Public Health Service policy on humane care and use of laboratory animals by awardee institutions; notice. Fed Regist 50:19584–19585Google Scholar
  15. Hokari M, Kuroda S, Shichinohe H, Yano S, Hida K, Iwasaki Y (2008) Bone marrow stromal cells protect and repair damaged neurons through multiple mechanisms. J Neurosci Res 86:1024–1035. CrossRefGoogle Scholar
  16. Horita Y, Honmou O, Harada K, Houkin K, Hamada H, Kocsis JD (2006) Intravenous administration of glial cell line-derived neurotrophic factor gene-modified human mesenchymal stem cells protects against injury in a cerebral ischemia model in the adult rat. J Neurosci Res 84:1495–1504. CrossRefGoogle Scholar
  17. Hu GQ, Du X, Li YJ, Gao XQ, Chen BQ, Yu L (2017) Inhibition of cerebral ischemia/reperfusion injury-induced apoptosis: nicotiflorin and JAK2/STAT3 pathway. Neural Regen Res 12:96–102. CrossRefGoogle Scholar
  18. Lehmann U, Schmitz J, Weissenbach M, Sobota RM, Hörtner M, Friederichs K, Behrmann I, Tsiaris W, Sasaki A, Schneider-Mergener J, Yoshimura A, Neel BG, Heinrich PC, Schaper F (2003) SHP2 and SOCS3 contribute to Tyr-759-dependent attenuation of interleukin-6 signaling through gp130. J Biol Chem 278:661–671. CrossRefGoogle Scholar
  19. Li Y, Chen J, Chen XG, Wang L, Gautam SC, Xu YX, Katakowski M, Zhang LJ, Lu M, Janakiraman N, Chopp M (2002) Human marrow stromal cell therapy for stroke in rat: neurotrophins and functional recovery. Neurology 59:514–523CrossRefGoogle Scholar
  20. Luan Y, Zhang X, Zhang Y, Dong Y (2017) MicroRNA-210 protects PC-12 cells against hypoxia-induced injury by targeting BNIP3. Front Cell Neurosci 11:285. CrossRefGoogle Scholar
  21. Matsushita K, Itoh S, Ikeda S, Yamamoto Y, Yamauchi Y, Hayashi M (2014) LIF/STAT3/SOCS3 signaling pathway in murine bone marrow stromal cells suppresses osteoblast differentiation. J Cell Biochem 115:1262–1268. CrossRefGoogle Scholar
  22. Mishra KK, Gupta S, Banerjee K (2016) SOCS3 induces neurite differentiation and promotes neuronal cell survival. IUBMB Life 68:468–476. CrossRefGoogle Scholar
  23. Pimentel VC, Pinheiro FV, de Bona KS, Maldonado PA, da Silva CR, de Oliveira SM, Ferreira J, Bertoncheli CM, Schetinger MR, da Luz SCA, Moretto MB (2011) Hypoxic-ischemic brain injury stimulates inflammatory response and enzymatic activities in the hippocampus of neonatal rats. Brain Res 1388:134–140. CrossRefGoogle Scholar
  24. Raghavendra Rao VL, Bowen KK, Dhodda VK, Song G, Franklin JL, Gavva NR, Dempsey RJ (2002) Gene expression analysis of spontaneously hypertensive rat cerebral cortex following transient focal cerebral ischemia. J Neurochem 83:1072–1086CrossRefGoogle Scholar
  25. Takano K, Ogura M, Nakamura Y, Yoneda Y (2005) Neuronal and glial responses to polyamines in the ischemic brain. Curr Neurovasc Res 2:213–223CrossRefGoogle Scholar
  26. Tondreau T, Lagneaux L, Dejeneffe M, Massy M, Mortier C, Delforge A, Bron D (2004) Bone marrow-derived mesenchymal stem cells already express specific neural proteins before any differentiation. Differentiation 72:319–326. CrossRefGoogle Scholar
  27. Tso D, McKinnon RD (2015) Cell replacement therapy for central nervous system diseases. Neural Regen Res 10:1356–1358. CrossRefGoogle Scholar
  28. van de Geijn GJ, Gits J, Aarts LH, Heijmans-Antonissen C, Touw IP (2004) G-CSF receptor truncations found in SCN/AML relieve SOCS3-controlled inhibition of STAT5 but leave suppression of STAT3 intact. Blood 104:667–674. CrossRefGoogle Scholar
  29. Wang Y, Deng Y, Zhou GQ (2008) SDF-1alpha/CXCR4-mediated migration of systemically transplanted bone marrow stromal cells towards ischemic brain lesion in a rat model. Brain Res 1195:104–112. CrossRefGoogle Scholar
  30. Wang Z, Liu T, Gan L, Wang T, Yuan X, Zhang B, Chen H, Zheng Q (2010) Shikonin protects mouse brain against cerebral ischemia/reperfusion injury through its antioxidant activity. Eur J Pharmacol 643:211–217. CrossRefGoogle Scholar
  31. Wang X, Wang XL, Chen HL, Wu D, Chen JX, Wang XX, Li RL, He JH, Mo L, Cen X, Wei YQ, Jiang W (2014) Ghrelin inhibits doxorubicin cardiotoxicity by inhibiting excessive autophagy through AMPK and p38-MAPK. Biochem Pharmacol 88:334–350. CrossRefGoogle Scholar
  32. Wang XL, Qiao CM, Liu JO, Li CY (2017) Inhibition of the SOCS1-JAK2-STAT3 signaling pathway confers neuroprotection in rats with ischemic stroke. Cell Physiol Biochem 44:85–98. CrossRefGoogle Scholar
  33. Wislet-Gendebien S, Wautier F, Leprince P, Rogister B (2005) Astrocytic and neuronal fate of mesenchymal stem cells expressing nestin. Brain Res Bull 68:95–102. CrossRefGoogle Scholar
  34. Wu D, Lu W, Wei Z, Xu M, Liu X (2018) Corrigendum to “neuroprotective effect of Sirt2-specific inhibitor AK-7 against acute cerebral ischemia is P38 activation-dependent in mice” [Neuroscience 374 (2018) 61-69]. Neuroscience 382:154–156. CrossRefGoogle Scholar
  35. Yamaguchi M, Okamoto K, Kusano T, Matsuda Y, Suzuki G, Fuse A, Yokota H (2015) The effects of xanthine oxidoreductase inhibitors on oxidative stress markers following global brain ischemia reperfusion injury in C57BL/6 mice. PLoS One 10:e0133980. CrossRefGoogle Scholar
  36. Zhang H, Huang Z, Xu Y, Zhang S (2006a) Differentiation and neurological benefit of the mesenchymal stem cells transplanted into the rat brain following intracerebral hemorrhage. Neurol Res 28:104–112. CrossRefGoogle Scholar
  37. Zhang L, Badgwell DB, Bevers JJ 3rd, Schlessinger K, Murray PJ, Levy DE, Watowich SS (2006b) IL-6 signaling via the STAT3/SOCS3 pathway: functional analysis of the conserved STAT3 N-domain. Mol Cell Biochem 288:179–189. CrossRefGoogle Scholar
  38. Zheng Y, Wu Z, Yi F, Orange M, Yao M, Yang B, Liu J, Zhu H (2018) By activating Akt/eNOS bilobalide B Inhibits Autophagy and promotes angiogenesis following focal cerebral ischemia reperfusion. Cell Physiol Biochem 47:604–616. CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Rehabilitation MedicineLinyi Central HospitalLinyiChina
  2. 2.Department of GynaecologyLinyi Central HospitalLinyiChina
  3. 3.Department of InfectionLinyi Central HospitalLinyiChina

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