Published online Mar 7, 2006. doi: 10.3748/wjg.v12.i9.1392
Revised: November 1, 2005
Accepted: November 10, 2005
Published online: March 7, 2006
AIM: To study the relationship between interleukin-1beta (IL-1β) up-regulating tissue inhibitor of matrix metalloproteinase-1 (TIMMP-1) mRNA expression and phosphorylation of both c-jun N-terminal kinase (JNK) and p38 in rat hepatic stellate cells (HSC).
METHODS: RT-PCR was performed to measure the expression of TIMMP-1 mRNA in rat HSC. Western blot was performed to measure IL-1β-induced JNK and p38 activities in rat HSC.
RESULTS: TIMMP-1 mRNA expression (1.191 ± 0.079) was much higher after treatment with IL-1β (10 ng/mL) for 24 h than in control group (0.545 ± 0.091) (P<0.01). IL-1β activated JNK and p38 in a time-dependent manner. After stimulation with IL-1β for 0, 5, 15, 30, 60 and 120 min，the JNK activity was 0.982 ± 0.299，1.501 ± 0.720, 2.133 ± 0.882, 3.360 ± 0.452, 2.181 ± 0.789, and 1.385 ± 0.368, respectively. There was a significant difference in JNK activity at 15 min (P < 0.01), 30 min (P < 0.01) and 60 min (P < 0.01) in comparison to that at 0 min. The p38 activity was 1.061 ± 0.310，2.050 ± 0.863，2.380 ± 0.573, 2.973 ± 0.953, 2.421 ± 0.793, and 1.755 ± 0.433 at the 6 time points (0, 5, 15, 30, 60 and 120 min) respectively. There was a significant difference in p38 activity at 5 min (P < 0.05), 15 min (P < 0.01), 30 min (P < 0.01) and 60 min (P < 0.01) compared to that at 0 min. TIMMP-1 mRNA expression trended to decrease in 3 groups pretreated with different concentrations of SP600125 (10 µmol/L, 1.022 ± 0.113; 20 µmol/L, 0.869 ± 0.070; 40 µmol/L, 0.666 ± 0.123). Their decreases were all significant (P < 0.05, P < 0.01, P < 0.01) in comparison to control group (without SP600125 treatment, 1.163 ± 0.107). In the other 3 groups pretreated with different concentrations of SB203580 (10 µmol/L, 1.507 ± 0.099; 20 µmol/L, 1.698 ± 0.107; 40 µmol/L, 1.857 ± 0.054), the expression of TIMMP-1 mRNA increased. Their levels were higher than those in the control group (without SB203580 treatment, 1.027 ± 0.061) with a significant statistical significance (P < 0.01).
CONCLUSION: IL-1β has a direct action on hepatic fibrosis by up-regulating TIMMP-1 mRNA expression in rat HSC. JNK and p38 mitogen-activated protein kinases (MAPKs) are involved in IL-1β-induced TIMMP-1 gene expression, and play a distinct role in this process, indicating that p38 and JNK pathways cooperatively mediate TIMP-1 mRNA expression in rat HSC.