Expression of inducible nitric oxide synthase in the liver is under the control of nuclear factor kappa B in concanavalin A-induced hepatitis.

OBJECTIVE

Both nuclear factor kappa B (NF-kappaB) activation and inducible nitric oxide synthase (iNOS) expression increase in the liver injury, and there are NF-kappaB binding sites in the iNOS promoter. The aim of this study was to investigate the correlation between iNOS expression and NF-kappaB activation in hepatitis induced by concanavalin A (con A).

METHODS

Eighty-eight male BALB/c mice were randomly divided into three groups: vehicle control group, con A group and pyrrolidine dithiocarbamate (PDTC) plus con A group. In the vehicle control group, the mice were treated with saline (0.3 mL, i.v.). In the con A group, the mice were treated with con A (20 mg/kg, i.v.). In the PDTC + con A group, the mice were pretreated with PDTC (120 mg/kg, i.p.) 30 min before administration of con A (20 mg/kg, i.v.). Blood samples were taken from the retro-orbital venous plexus at 0.5, 1, 4, 8 and 16 h after con A injection and the mice were killed immediately. The plasma alanine aminotransferase (ALT) levels were measured by the standard photometric method. Nitric oxide (NO) levels in the liver homogenate were assayed by spectroscopy. Liver tissues were sectioned and stained with hematoxylin-eosin for histological examination. Activation of NF-kappaB, degradation of inhibitor of kappa B alpha (IkappaBalpha), and expression of iNOS were measured by western blot.

RESULTS

In the con A group, the plasma ALT activity and NO levels in the liver increased significantly at 1 h (P < 0.05, n = 8) and reached a peak at 4 h after con A injection. The liver injury in this group was characterized by liver necrosis, cell swelling and fatty degeneration. Cytosolic IkappaBalpha decreased slightly at 30 min after con A challenge, was undetectable at 1 h and reappeared at 4 h. Correspondingly, the NF-kappaB level in the nucleus was highest at 1 h. The iNOS expression increased at 30 min after con A injection and reached a maximum at 4 h. Pretreatment with PDTC prevented these changes and attenuated the liver injury.

CONCLUSIONS

Con A-induced iNOS expression in the liver is dependent on the activation of NF-kappaB.