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Fengyuan Piao
Affiliated Zhongshan Hospital of Dalian University
Corresponding Author
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Background: N-hexane, with its metabolite 2,5-hexanedine (HD), is a hazardous material widely used in industry and chronic exposure causes nerve demyelination. Demyelinating conditions are difficult to treat and frequently cause disabilities. Stem cell therapy using bone marrow mesenchymal stem cells (BMSCs) is a promising novel strategy. Our previous study found that BMSCs promoted significant recovery of motor dysfunction in rats modelling N-hexane neurotoxicity. The present study aimed to explore the underlying mechanisms and focused on the changes in spinal cord.
Methods: Sprague Dawley rats were intoxicated with HD (400 mg/kg/day, i.p, for 5 weeks). BMSCs (5×107cells/kg) were administrated by tail vein injection. Demyelination and remyelination of the spinal cord before and after BMSC treatment were examined microscopically. Cultured oligodendrocyte progenitor cells (OPCs) were incubated with HD +/- BMSCs-derived conditional medium (BMSC-CM). OPC differentiation was studied by immunostaining and quantitative imaging analysis. The expressional changes of Hes1, a key transcription factor negatively regulating OPC-differentiation, were studied. The upstream Notch1 and TNFα/RelB pathways were studied and some key signaling molecules in these pathways were measured. The correlation between neurotrophin NGF and TNFα was also investigated. Statistical significance was evaluated using one-way ANOVA test and performed using SPSS 13.0.
Results: The demyelinating damage by HD and remyelination by BMSCs were evidenced by electron microscopy, LFB staining and NG2/MBP immunohistochemistry. In vitro cultured OPCs showed more differentiation after incubation with BMSC-CM. Hes1 expression was found to be significantly increased by HD and decreased by BMSC or BMSC-CM. The change of Hes1 was found, however, independent on Notch1 activation, but dependent on TNFα/RelB signaling. HD was found to increase TNFα, RelB and Hes1 expression and BMSCs was found to have the opposite effect. Addition of recombinant TNFα to OPCs or RelB overexpression similarly caused upregulation of Hes1 expression. The secretion of NGF by BMSC and activation of NGF receptor was found important for suppression of TNFα production in OPCs.
Conclusions: Our findings demonstrated that BMSCs promote remyelination in the spinal cord of HD-exposed rats via TNFα/RelB-Hes1 pathway, providing novel insights for evaluating and further exploring the therapeutical effect of BMSCs on demyelinating neurodegenerative disease.
Keywords
bone marrow-mesenchymal stem cells, 2,5-hexanedione, demyelination, remyelination, oligodendrocyte precursor cells, Notch1, TNFα, RelB, Hes1, NGF
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View the published article at Stem Cell Research & Therapy.
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Posted 26 Jun, 2021
Editorial decision: Minor Revision
On 26 Jun, 2021
Editor assigned
On 13 Jun, 2021
Submission checks complete
On 13 Jun, 2021
Editor invited
On 13 Jun, 2021
First submitted
On 10 Jun, 2021
This version was not preprinted
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Posted 13 Apr, 2021
This version was not preprinted
Version 1
Posted 13 Apr, 2021
No community comments so far
Editorial decision: Major Revision
On 15 May, 2021
Review #2 received
Received 06 May, 2021
Reviewer #2 agreed
On 25 Apr, 2021
Reviews received
Received 25 Apr, 2021
Review #1 received
Received 23 Apr, 2021
Reviewer #1 agreed
On 12 Apr, 2021
Reviewers invited
Invitations sent on 10 Apr, 2021
Editor assigned
On 07 Apr, 2021
Submission checks complete
On 06 Apr, 2021
Editor invited
On 06 Apr, 2021
First submitted
On 06 Apr, 2021
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See the published version of this preprint at Stem Cell Research & Therapy.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Fengyuan Piao
Affiliated Zhongshan Hospital of Dalian University
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at Stem Cell Research & Therapy.
Version (no preprint)
Posted 26 Jun, 2021
Editorial decision: Minor Revision
On 26 Jun, 2021
Editor assigned
On 13 Jun, 2021
Submission checks complete
On 13 Jun, 2021
Editor invited
On 13 Jun, 2021
First submitted
On 10 Jun, 2021
This version was not preprinted
Version (no preprint)
Posted 13 Apr, 2021
This version was not preprinted
Version 1
Posted 13 Apr, 2021
No community comments so far
Editorial decision: Major Revision
On 15 May, 2021
Review #2 received
Received 06 May, 2021
Reviewer #2 agreed
On 25 Apr, 2021
Reviews received
Received 25 Apr, 2021
Review #1 received
Received 23 Apr, 2021
Reviewer #1 agreed
On 12 Apr, 2021
Reviewers invited
Invitations sent on 10 Apr, 2021
Editor assigned
On 07 Apr, 2021
Submission checks complete
On 06 Apr, 2021
Editor invited
On 06 Apr, 2021
First submitted
On 06 Apr, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Stem Cell Research & Therapy.
Background: N-hexane, with its metabolite 2,5-hexanedine (HD), is a hazardous material widely used in industry and chronic exposure causes nerve demyelination. Demyelinating conditions are difficult to treat and frequently cause disabilities. Stem cell therapy using bone marrow mesenchymal stem cells (BMSCs) is a promising novel strategy. Our previous study found that BMSCs promoted significant recovery of motor dysfunction in rats modelling N-hexane neurotoxicity. The present study aimed to explore the underlying mechanisms and focused on the changes in spinal cord.
Methods: Sprague Dawley rats were intoxicated with HD (400 mg/kg/day, i.p, for 5 weeks). BMSCs (5×107cells/kg) were administrated by tail vein injection. Demyelination and remyelination of the spinal cord before and after BMSC treatment were examined microscopically. Cultured oligodendrocyte progenitor cells (OPCs) were incubated with HD +/- BMSCs-derived conditional medium (BMSC-CM). OPC differentiation was studied by immunostaining and quantitative imaging analysis. The expressional changes of Hes1, a key transcription factor negatively regulating OPC-differentiation, were studied. The upstream Notch1 and TNFα/RelB pathways were studied and some key signaling molecules in these pathways were measured. The correlation between neurotrophin NGF and TNFα was also investigated. Statistical significance was evaluated using one-way ANOVA test and performed using SPSS 13.0.
Results: The demyelinating damage by HD and remyelination by BMSCs were evidenced by electron microscopy, LFB staining and NG2/MBP immunohistochemistry. In vitro cultured OPCs showed more differentiation after incubation with BMSC-CM. Hes1 expression was found to be significantly increased by HD and decreased by BMSC or BMSC-CM. The change of Hes1 was found, however, independent on Notch1 activation, but dependent on TNFα/RelB signaling. HD was found to increase TNFα, RelB and Hes1 expression and BMSCs was found to have the opposite effect. Addition of recombinant TNFα to OPCs or RelB overexpression similarly caused upregulation of Hes1 expression. The secretion of NGF by BMSC and activation of NGF receptor was found important for suppression of TNFα production in OPCs.
Conclusions: Our findings demonstrated that BMSCs promote remyelination in the spinal cord of HD-exposed rats via TNFα/RelB-Hes1 pathway, providing novel insights for evaluating and further exploring the therapeutical effect of BMSCs on demyelinating neurodegenerative disease.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
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