Effects of recombinant adeno-associated virus mediated nerve growth factor gene transfection on oligodendrocyte apoptosis and myelination in experimental autoimmune encephalomyelitis mice

doi:10.12307/2021.039

Abstract

Abstract: BACKGROUND: Nerve growth factor (NGF) has an inhibitory effect on normal neuronal apoptosis, thereby improving the ability to repair cell damage, and playing a therapeutic role in some autoimmune diseases.
OBJECTIVE: To observe the effect of recombinant adeno-associated virus mediated nerve growth factor (Ad-NGF) on apoptosis and myelination of oligodendrocytes in experimental autoimmune encephalomyelitis (EAE) mice.
METHODS: Thirty female healthy C57BL/6 mice were randomly divided into normal group, EAE group and transfection group, with 10 mice in each group. In the EAE group and transfection group, EAE models were made in mice using myelin oligodendrocyte glycoprotein peptide immunoassay. Three days after modeling, mice in the EAE group were injected normal saline via the tail vein for 21 continuous days, while those in the transfection group were injected Ad-NGF via the tail vein for 21 continuous days. All the mice were executed at the peak period of the disease. LFB staining was used to observe the morphology and pathology of myelin tissue. Immunofluorescence method was used to observe the expression and co-localization of apoptotic protein Caspase3 and oligodendrocytes in spinal cord tissue. RT-PCR method was used to detect the mRNA levels of NGF and myelin alkali in spinal cord tissue. Western blot assay was used to detect the protein levels of NGF and Caspase3 in spinal cord tissue. ELISA was used to measure the level of myelin basic protein in spinal cord tissue. The study protocol was approved by the Animal Ethics Committee of Southwest Medical University (approval No. 201912-8).
RESULTS AND CONCLUSION: LFB staining showed significant demyelination changes in the EAE group, while the demyelination was significantly improved in the transfection group. Caspase-3 aggregation was obviously observed in oligodendrocytes of EAE group, but not in transfection group. RT-PCR results indicated that the mRNA levels of myelin basic protein and NGF were significantly lower in the EAE group than the normal control and transfection groups (P < 0.05). Western blot results revealed that in the EAE group the level of Caspase3 protein was significantly increased (P < 0.05), while the level of NGF significantly reduced as compared with the normal control and transfection groups (P < 0.05). ELISA results showed that the level of myelin basic protein in the EAE group was significantly lower than those in the normal control and transfection groups (P < 0.05). To conclude, the Ad-NGF transfected by external turnover has preventive and control effects on the EAE mouse model, and its mechanism may be related to upregulation of NGF level, down-regulation of Caspase3 in oligodendrocytes, and promotion of myelin basic protein expression, thereby improving demyelination.

Key words: experiment, animal, mouse, nerve growth factor, adenovirus, autoimmune encephalomyelitis, oligodendrocyte, myelin sheath, apoptosis, protein

CLC Number:

Xie Yang, Lü Zhiyu, Zhang Shujiang, Long Ting, Li Zuoxiao. Effects of recombinant adeno-associated virus mediated nerve growth factor gene transfection on oligodendrocyte apoptosis and myelination in experimental autoimmune encephalomyelitis mice[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(23): 3678-3683.

Figures/Tables 6

References

[1] SOSPEDRA M, MARTIN R. Immunology of Multiple Sclerosis. Semin Neurol. 2016;36(2):115-127.
[2] ZHOU X, LI X, FENG M, et al. Analysis of the direct injury effector of oligodendroglia cells or myelin sheath in an experimental allergic encephalomyelitis model induced by the MOG35-55 peptide. Mol Med Rep. 2015;27(8):124-131.
[3] LI Q, HOUDAYER T, LIU S, et al. Induced neural activity promotes an oligodendroglia regenerative response in the injured spinal cord and improves motor function after spinal cord injury. J Neurotrauma. 2017; 34(24):3351-3361.
[4] DULAMEA AO. The contribution of oligodendrocytes and oligodendrocyte progenitor cells to central nervous system repair in multiple sclerosis: perspectives for remyelination therapeutic strategies. Neural Regen Res. 2017;12(12):1939-1944.
[5] GHASEMI N. The Evaluation of Astaxanthin Effects on Differentiation of Human Adipose Derived Stem Cells into Oligodendrocyte Precursor Cells. Avicenna J Med Biotechnol. 2018;10(2):69-74.
[6] CARDOUAT G, DOUARD M, ROBILLARD P, et al. Short-term mechanisms activated by the nerve growth factor NGF to induce pulmonary arterial hyperreactivity. Arch Cardiovasc Dis. 2019;11(2):045.
[7] CHI J, SONG WZ, FAN QS, et al. Transfection characters of NGF recombination adenoviral in spiral ganglion cells. Med J National. 2015; 11(1):1004-0188.
[8] DE PAULA FARIA D. Myelin positron emission tomography (PET) imaging in multiple sclerosis. Neural Regen Res. 2020;15(10):1842-1843.
[9] SKARLIS C, ANAGNOSTOULI M. The role of melatonin in Multiple Sclerosis. Neurol Sci. 2020;41(4):769-781.
[10] TALEB HAA, ALGHAMDI BS. Neuroprotective Effects of Melatonin during Demyelination and Remyelination Stages in a Mouse Model of Multiple Sclerosis. J Mol Neurosci. 2019;70(3):386-402.
[11] STEINMAN L. Inverse Vaccination to Silence Immunity to Myelin in Multiple Sclerosis. Can J Neurol Sci. 2010; 37(2):49-58.
[12] DE PAULA FARIA D. Myelin positron emission tomography (PET) imaging in multiple sclerosis. Neural Regen Res. 2020;15(10):1842-1843.
[13] LISAK RP, BENJAMINS JA, NEDELKOSKA L, et al. Secretory products of multiple sclerosis B cells are cytotoxic to oligodendroglia in vitro. J Neuroimmunol. 2012;246(1-2):85-95.
[14] NOTTINGHAM S, KNAPP P, SPRINGER J. FK506 Treatment Inhibits Caspase-3 Activation and Promotes Oligodendroglial Survival Following Traumatic Spinal Cord Injury. Exp Neurol. 2002;177(1):242-251.
[15] OLFA R, MOHAMED A. Chlorogenic Acid Prevents AMPA-Mediated Excitotoxicity in Optic Nerve Oligodendrocytes Through a PKC and Caspase-Dependent Pathways. Neurotox Res. 2018;13(4):104-129.
[16] DAS A, GUYTON M, BUTLER J, et al. Activation of Calpain and Caspase Pathways in Demyelination and Neurodegeneration in Animal Model of Multiple Sclerosis. CNS Neurol Disord Drug Targets. 2008;7(3):313-320.
[17] DENG C, LI J, LI L, et al. Effects of hypoxia ischemia on caspase-3 expression and neuronal apoptosis in the brain of neonatal mice. Exp Ther Med. 2019;17(6):4517.
[18] LI R, LI YY, LI DH, et al. NGF Promotes Myelin Debris Clearance after Peripheral Nerve Injury by Inhibiting Endoplasmic Reticulum Stress. Chin J Biochem Mol Biol. 2018;5(2):89-96.
[19] KANG WB, CHEN YJ, LU DY, et al. Folic acid contributes to peripheral nerve injury repair by promoting Schwann cell proliferation, migration, and secretion of nerve growth factor. Neural Regen Res. 2019;14(1):132-139.
[20] MIR S, ALI F, CHAUHAN D, et al. Accumulation of reactivity to MBP sensitizes TRAIL mediated oligodendrocyte apoptosis in adult sub cortical white matter in a model for human multiple sclerosis. Metab Brain Dis. 2016;31(2):299-309.
[21] YE JL, CHEN XS, SU L, et al. Progesterone Alleviates Neural Behavioral Deficits and Demyelination with Reduced Degeneration of Oligodendroglial Cells in Cuprizone-Induced Mice. PLoS One. 2013; 8(1):54590-54621.
[22] STEPANOV AV, BELOGUROV AA, MAMEDOV AÉ, et al. Therapeutic effect of encapsulated into the nanocontainers MBP immunodominant peptides on EAE development in DA rats. Bioorg Khim. 2012;38(3): 306-314.
[23] AULOVA KS, TOPORKOVA LB, LOPATNIKOVA JA, et al. Changes in haematopoietic progenitor colony differentiation and proliferation and the production of different abzymes in EAE mice treated with DNA.J Cell Mol Med. 2017;42(2):109-123.
[24] LEI J. Evolutionary dynamics of cancer: From epigenetic regulation to cell population dynamics—mathematical model framework, applications, and open problems. Ence China Mathematics. 2020;63(3):411-424.
[25] SHCHEPAREVA ME, KOCHERGIN IA, TOLPEEVA OA, et al. Diagnostic value of antibodies to myelin oligodendrocyte glycoprotein in demyelinating diseases of the central nervous system. Zh Nevrol Psikhiatr Im S S Korsakova. 2019;119(2):18-23.