Zhang Ya-Ping, Yao Xi-Xian, Zhao Xia
Department of Gastroenterology, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China.
World J Gastroenterol. 2006 Mar 7;12(9):1392-6. doi: 10.3748/wjg.v12.i9.1392.
To study the relationship between interleukin-1beta (IL-1beta) 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).
RT-PCR was performed to measure the expression of TIMMP-1 mRNA in rat HSC. Western blot was performed to measure IL-1beta-induced JNK and p38 activities in rat HSC.
TIMMP-1 mRNA expression (1.191+/-0.079) was much higher after treatment with IL-1beta (10 ng/mL) for 24 h than in control group (0.545+/-0.091) (P<0.01). IL-1beta activated JNK and p38 in a time-dependent manner. After stimulation with IL-1beta 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 micromol/L, 1.022+/-0.113; 20 micromol/L, 0.869+/-0.070; 40 micromol/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 micromol/L, 1.507+/-0.099; 20 micromol/L, 1.698+/-0.107; 40 micromol/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).
IL-1beta 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-1beta-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.
研究白细胞介素-1β（IL-1β）上调大鼠肝星状细胞（HSC）中基质金属蛋白酶组织抑制因子-1（TIMMP-1）mRNA表达与c-jun氨基末端激酶（JNK）和p38磷酸化之间的关系。
采用逆转录聚合酶链反应（RT-PCR）检测大鼠HSC中TIMMP-1 mRNA的表达。采用蛋白质免疫印迹法检测IL-1β诱导的大鼠HSC中JNK和p38的活性。
用IL-1β（10 ng/mL）处理24 h后，TIMMP-1 mRNA表达（1.191±0.079）显著高于对照组（0.545±0.091）（P<0.01）。IL-1β以时间依赖性方式激活JNK和p38。用IL-1β刺激0、5、15、30、60和120分钟后，JNK活性分别为0.982±0.299、1.501±0.720、2.133±0.882、3.360±0.452、2.181±0.789和1.385±0.368。与0分钟时相比，15分钟（P<0.01）、30分钟（P<0.01）和60分钟（P<0.01）时JNK活性有显著差异。6个时间点（0、5、15、30、60和120分钟）的p38活性分别为1.061±0.310、2.050±0.863、2.380±0.573、2.973±0.953、2.421±0.793和1.755±0.433。与0分钟时相比，5分钟（P<0.05）、15分钟（P<0.01）、30分钟（P<0.01）和60分钟（P<0.01）时p38活性有显著差异。用不同浓度的SP600125（10 μmol/L，1.022±0.113；20 μmol/L，0.869±0.070；40 μmol/L，0.666±0.123）预处理的3组中，TIMMP-1 mRNA表达呈下降趋势。与对照组（未用SP600125处理，1.163±0.107）相比，其下降均有统计学意义（P<0.05，P<0.01，P<0.01）。在另外3组用不同浓度SB203580（10 μmol/L，1.507±0.099；20 μmol/L，1.698±0.107；40 μmol/L，1.857±0.054）预处理的组中，TIMMP-1 mRNA表达增加。其水平高于对照组（未用SB203580处理，1.027±
