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Expression of functional recombinant human fibroblast growth factor 8b and its protective effects on MPP+-lesioned PC12 cells

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Abstract

Human fibroblast growth factor 8b (FGF8b) was expressed based on a baculovirus expression vector system (BEVS) and identified as having a protective effect on Parkinson’s disease. Immunoblotting demonstrated that rhFGF8b proteins were recognized by a human anti-FGF8b antibody. The multiplicity of infection and timing of harvest had a significant effect on protein yield and protein quality. Our results indicated that the rhFGF8b was first detectable at 36 h postinfection and reached a maximum at 60 h. A multiplicity of infection (MOI) of 8 pfu/mL was suitable for harvest. The target protein was purified by heparin-affinity chromatography. In vitro methylthiazol tetrazolium (MTT) assays demonstrated that the purified rhFGF8b could significantly stimulate proliferation of NIH3T3 cells. Furthermore, to elucidate the effect of rhFGF8b on Parkinson’s disease, we used FGF8b pretreatment on a cell model of Parkinson’s disease. The results indicated that rhFGF8b prevented necrosis and apoptosis of 1-METHYL-4-phenyl pyridine (MPP+) treated PC12 cells. Moreover, the effect of FGF8b on messenger RNA (mRNA) levels of apoptosis and ERS genes was investigated to clarify the molecular mechanisms of FGF8b. The results suggest that FGF8b exerts neuroprotective effects by alleviating endoplasmic reticulum (ER) stress during PD. These results suggest that FGF8b may be a promising candidate therapeutic drug for neurodegenerative diseases related to ER stress.

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Acknowledgments

The project was supported by Zhejiang Extremely Key Subject of Pharmacology and Biochemical Pharmaceutics and the National “863” High Technology Research and Development Program (2011AA02A113).

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The authors declare that they have no competing interests.

This article does not contain any studies with human participants performed by any of the authors.

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Informed consent was obtained from all individual participants included in the study.

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Authors and Affiliations

  1. School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China

    Nazi Chen, Jishen Ma, Yang Zhao, Meiyu Wu, Huanhuan Yang, Weiyue Gong, Jiang Chao & Xiaokun Li

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  1. Nazi Chen
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  2. Jishen Ma
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  3. Yang Zhao
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Correspondence to Jiang Chao or Xiaokun Li.

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Chen, N., Ma, J., Zhao, Y. et al. Expression of functional recombinant human fibroblast growth factor 8b and its protective effects on MPP+-lesioned PC12 cells. Appl Microbiol Biotechnol 100, 625–635 (2016). https://doi.org/10.1007/s00253-015-7004-4

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