Effects of recombinant human thioredox on primary cultured hepatocytes
- VernacularTitle:重组人硫氧化还原蛋白对原代培养肝细胞的影响
- Author:
Hua LI
;
Genshu WANG
;
Guihua CHEN
;
Minqiang LU
;
Yang YANG
;
Changjie CAI
;
Chi XU
;
Shuhong YI
- Publication Type:Journal Article
- Keywords:
Hepatocytes;
Gene expression;
Human thioredoxin
- From:
Chinese Journal of General Surgery
2000;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of recombinant human thioredox ( rhTrx) on primary cultured hepatocytes. Methods The cDNA coding hTrx was amplified by using RT-PCR. The objective protein was expressed in E. Coli. The activity of purified rhTrx protein was determined by IgM and insulin disulfide reduction assay. DNA synthesis was determined by 3H-TdR mixing assay to realize the proliferating roles of rhTrx in murine primary cultured hepatocytes. Cell viability was assessed by lactate dehydrogenase ( LDH) releasing assay. Results The cloning hTrx open reading frames cDNA was confirmed by sequence analysis. The soluble recombinant hTrx are expressed efficiently in the prokaryotic expression system. The recovery ratio was 23. 20% of total bacteria protein with a purity of the expressive protein of 96. 30%. The rhTrx had stronger IgM and insulin reduced activity compared with the controls (P
