Abstract
In recent years, an increasing number of studies have shown that fibroblast growth factor 12 (FGF12) plays important roles in regulating neural development and function. Importantly, changes of FGF12 expression are thought to be related to the pathophysiology of many neurological diseases. However, little research has been performed to explore the protective effect of FGF12 on nerve damage. This study aims to explore its neuroprotective effects using our recombinant humanized FGF12 (rhFGF12). The hFGF12 gene was cloned and ligated into an expression vector to construct a recombinant plasmid pET-3a-hFGF12. Single colonies were screened to obtain high expression engineering strains, and fermentation and purification protocols for rhFGF12 were designed and optimized. The biological activities and related mechanisms of rhFGF12 were investigated by MTT assay using NIH3T3 and PC12 cell lines. The in vitro neurotoxicity model of H2O2-induced oxidative injury in PC12 cells was established to explore the protective effects of rhFGF12. The results indicate that the beneficial effects of rhFGF12 were most likely achieved by promoting cell proliferation and reducing apoptosis. Moreover, a transgenic zebrafish (islet) with strong GFP fluorescence in the motor neurons of the hindbrain was used to establish a central injury model caused by mycophenolate mofetil (MMF). The results suggested that rhFGF12 could ameliorate central injury induced by MMF in zebrafish. In conclusion, we have established an efficient method to express and purify active rhFGF12 using an Escherichia coli expression system. Besides, rhFGF12 plays a protective effect of on nerve damage, and it provides a promising therapeutic approach for nerve injury.
Key points
• Effective expression and purification of bioactive rhFGF12 protein in E. coli.
• ERK/MAPK pathway is involved in rhFGF12-stimulated proliferation on PC12 cells.
• The rhFGF12 has the neuroprotective effects by inhibiting apoptosis.
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This work was funded by the Natural Science Foundation of Zhejiang Province (No. LY19H160029) and the Zhejiang Provincial Department of Education (No. Y201839837).
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HT and XL conceived and designed research. MZ, KM, YZ, MZ, PL, YF, and MH conducted experiments. JC and QD contributed new reagents or analytical tools. MZ and KM analyzed data. MZ and KM wrote the manuscript. All authors read and approved the manuscript.
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Zhou, M., Chen, J., Meng, K. et al. Production of bioactive recombinant human fibroblast growth factor 12 using a new transient expression vector in E. coli and its neuroprotective effects. Appl Microbiol Biotechnol 105, 5419–5431 (2021). https://doi.org/10.1007/s00253-021-11430-8
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DOI: https://doi.org/10.1007/s00253-021-11430-8