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Effective Protection on Acute Liver Injury by Halo Tag‐Flanked Recombinant Fibroblast Growth Factor 7
Author(s) -
Liu Min,
Chen Zhiyu,
Huang Sisi,
Chu Shenghui,
Issaro Nipatha,
Tian Haishan,
Hu Haiyan,
Jiang Chao
Publication year - 2018
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201700411
Subject(s) - recombinant dna , in vivo , apoptosis , western blot , in vitro , liver injury , microbiology and biotechnology , cell growth , cell culture , chemistry , protease , biology , pharmacology , biochemistry , gene , enzyme , genetics
The drug development of FGF7 has been restricted by its toxicity to the host, low expression, poor stability, and easy degradation. Recent studies have shown that Halo‐tag‐flanked recombinant human FGF7 can solve the problem of toxicity; however, its biological activity is unknown. This study aimed to explore the activity of Halo‐rhFGF7 and rhFGF7 on acute liver injury in vitro and in vivo. The rhFGF7 is expressed with a N‐terminal Halo‐tag, followed by a tobacco etch virus (TEV) protease cleavage site, in Escherichia coli BL21 (DE3) pLysS in this study. The products could stimulate the proliferation of carbon tetrachloride–damaged L‐O 2 cells (normal human liver cells); they also inhibited cell apoptosis. Due to the use of the Halo, the protein could be tracked using fluorescence localization. Recombinant protein exerted a protective effect on the acute liver injury model in vitro and in vivo. The MTT assay and Western blot analysis showed that this protective effect is realized through various paths, including promoting proliferation, inhibiting cell apoptosis and anti‐inflammatory. In conclusion, Halo‐rhFGF7 and rhFGF7 displayed an excellent protective effect on acute liver injury. The present study provided an experimental basis and data support for further research on rhFGF7.