title: Expression of functional recombinant human fibroblast growth factor 8b and its protective effects on MPP⁺-lesioned PC12 cells.
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.
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Nazi
authors.initials: N
authors.lastname: Chen
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Jishen
authors.initials: J
authors.lastname: Ma
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Meiyu
authors.initials: M
authors.lastname: Wu
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Yang
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Weiyue
authors.initials: W
authors.lastname: Gong
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China. chaojiang10@hotmail.com.
authors.forename: Jiang
authors.initials: J
authors.lastname: Chao
authors.affiliation: School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, 325035, China. proflxk@163.com.
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D016367:Baculoviridae / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002846:Chromatography, Affinity; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects; D051524:Fibroblast Growth Factor 8 / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D018696:Neuroprotective Agents / Q000302:isolation & purification / Q000494:pharmacology*; D016716:PC12 Cells; D010300:Parkinson Disease / Q000188:drug therapy / Q000503:physiopathology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology / Q000627:therapeutic use; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D018696:Neuroprotective Agents; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D051524:Fibroblast Growth Factor 8; C022616:thiazolyl blue
keywords: Baculovirus expression vector system; Endoplasmic reticulum (ER) stress; Fibroblast growth factor 8b; Neuroprotection; Parkinson’s disease