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

• Published in: Applied microbiology and biotechnology 2016-01, Vol.100 (2), p.625-635
• Main Authors: Chen, Nazi, Ma, Jishen, Zhao, Yang, Wu, Meiyu, Yang, Huanhuan, Gong, Weiyue, Chao, Jiang, Li, Xiaokun
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis - drug effects; Baculoviridae - genetics; Biomedical and Life Sciences; Biotechnological Products and Process Engineering; Biotechnology; Cell Proliferation - drug effects; Cell Survival - drug effects; Chromatography, Affinity; Endoplasmic Reticulum Stress - drug effects; Fibroblast Growth Factor 8 - biosynthesis; Fibroblast Growth Factor 8 - genetics; Fibroblast Growth Factor 8 - isolation & purification; Fibroblast Growth Factor 8 - pharmacology; Humans; Life Sciences; Mice; Microbial Genetics and Genomics; Microbiology; Neuroprotective Agents - isolation & purification; Neuroprotective Agents - pharmacology; NIH 3T3 Cells; Parkinson Disease - drug therapy; Parkinson Disease - physiopathology; PC12 Cells; Rats; Recombinant Proteins - isolation & purification; Recombinant Proteins - pharmacology; Recombinant Proteins - therapeutic use; RNA, Messenger - genetics; RNA, Messenger - metabolism; Tetrazolium Salts; Thiazoles
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-015-7004-4

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.