Stimulation of human hepatic stellate cells by cytochrome P4502E1-mediated oxidative stress / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 576-580, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-326293
ABSTRACT
<p><b>OBJECTIVE</b>To explore the stimulation of human hepatic stellate cells by Cytochrome P4502E1-mediated oxidative stress.</p><p><b>METHODS</b>HepG2-line was transfected with human CYP2E1 plasmid (HepG2/CYP2E1) and empty plasmid (HepG2/PCI) respectively. The CYP2E1 expression was evaluated with RT-PCR and Western blot. MDA was measured in culture medium of HepG2 cell lines. LX2 was co-incubated with HepG2/CYP2E1, HepG2/PCI and HepG2 respectively. The level of hydroxyproline in culture medium was examined in 48 hours and the cells were lysated and total RNA and protein were extracted. COL-1 and MMP2 mRNA levels were detected by RT-PCR and analyzed semi-quantitatively. PICP proteins were measured by ELISA. Zymography was performed to investigate MMP2 enzymatic activities.</p><p><b>RESULTS</b>(1) MDA from the HepG2 which (HepG2/CYP2E1)express human CYP2E1 (6.51+/-0.25) was significantly higher than that from the HepG2 which do not (HepG2/PCI) express human CYP2E1 (3.07+/-0.29) and HepG2 alone (2.57+/-0.29). (F=22.66, all P<0.01). (2) After co-incubated for 48 hours,the level of hydroxyproline in culture medium (35.24+/-3.52) excreted from CYP2E1/LX2 could significantly increase (F=58.89, P is less than 0.01). PICP protein (540.01+/-11.38) excreted from CYP2E1/LX2 was significantly increased (F=124.97, P<0.01). Zymography showed MMP2 gene expression and enzymatic activities of MMP2 had no difference among the groups (F=0.29, P>0.05) (F=0.33, P>0.05).</p><p><b>CONCLUSIONS</b>CYP2E1 derived oxidative stress mediated stimulation of collagen I synthesis by hepatic stellate cells. Hydroxyproline excreted by LX2 was increased by CYP2E1. COL-1mRNA had no difference among the groups (F=0.73, P>0.05).</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Oxidative Stress
/
Cytochrome P-450 CYP2E1
/
Bodily Secretions
/
Matrix Metalloproteinase 2
/
Collagen Type I
/
Hepatic Stellate Cells
/
Hep G2 Cells
/
Hydroxyproline
/
Liver Cirrhosis
/
Metabolism
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Hepatology
Year:
2010
Type:
Article
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