AIM: To examine the expressions of matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of metalloproteinases-1 (TINP-1) in rat fibrotic liver and in normal rat hepatic stellate cells, and to investigate the chang...AIM: To examine the expressions of matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of metalloproteinases-1 (TINP-1) in rat fibrotic liver and in normal rat hepatic stellate cells, and to investigate the changes in their expressions in response to treatment with interleukin-10 (IL-10) and platelet-derived growth factor (PDGF).METHODS: Rat models of CCl4-induced hepatic fibrosis were established and the liver tissues were sampled from the rats with or without IL-10 treatment, and also from the control rats. The expressions of MMP-2 and TIMP-1 in liver tissues were detected by S-P immunohistochemistry, and their expression intensities were evaluated in different groups. Hepatic stellate cells (HSCs) were isolated from normal rat and cultured in vitro prior to exposure to PDGF treatment or co-treatment with IL-10 and PDGF. MMP-2 and TIMP-1 levels were measured by semi-quantitative reverse transcriptional polymerase chain reaction (RT-PCR).RESULTS: CCl4- induced rat hepatic fibrosis models were successfully established. The positive expressions of MMP-2 and TIMP-1 increased obviously with the development of hepatic fibrosis, especially in untreated model group (84.0% and 92.0%, P<0.01). The positive signals decreased significantly following IL-10 treatment (39.3% and 71.4%,P<0.01 and P<0.05) in a time-dependent manner. TIMP-1 mRNA in PDGF-treated group was significantly increased time-dependently in comparison with that of the control group, but PDGF did not obviously affect MMP-2 expression.No difference was noted in TIMP-1 and MMP-2 expressions in HSCs after IL-10 and PDGF treatment (P>0.05).CONCLUSION: MMP-2 and TIMP-1 expressions increase in liver tissues with the development of fibrosis, which can be inhibited by exogenous IL-10 inhibitor. PDGF induces the up-regulation of TIMP-1 but not MMP-2 in the HSCs.IL-10 inhibits TIMP-1 and MMP-2 expressions in HSCs induced by PDGF.展开更多
基金Supported by the'Science and Technology Fund of Fujian Province,No.2003D05
文摘AIM: To examine the expressions of matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of metalloproteinases-1 (TINP-1) in rat fibrotic liver and in normal rat hepatic stellate cells, and to investigate the changes in their expressions in response to treatment with interleukin-10 (IL-10) and platelet-derived growth factor (PDGF).METHODS: Rat models of CCl4-induced hepatic fibrosis were established and the liver tissues were sampled from the rats with or without IL-10 treatment, and also from the control rats. The expressions of MMP-2 and TIMP-1 in liver tissues were detected by S-P immunohistochemistry, and their expression intensities were evaluated in different groups. Hepatic stellate cells (HSCs) were isolated from normal rat and cultured in vitro prior to exposure to PDGF treatment or co-treatment with IL-10 and PDGF. MMP-2 and TIMP-1 levels were measured by semi-quantitative reverse transcriptional polymerase chain reaction (RT-PCR).RESULTS: CCl4- induced rat hepatic fibrosis models were successfully established. The positive expressions of MMP-2 and TIMP-1 increased obviously with the development of hepatic fibrosis, especially in untreated model group (84.0% and 92.0%, P<0.01). The positive signals decreased significantly following IL-10 treatment (39.3% and 71.4%,P<0.01 and P<0.05) in a time-dependent manner. TIMP-1 mRNA in PDGF-treated group was significantly increased time-dependently in comparison with that of the control group, but PDGF did not obviously affect MMP-2 expression.No difference was noted in TIMP-1 and MMP-2 expressions in HSCs after IL-10 and PDGF treatment (P>0.05).CONCLUSION: MMP-2 and TIMP-1 expressions increase in liver tissues with the development of fibrosis, which can be inhibited by exogenous IL-10 inhibitor. PDGF induces the up-regulation of TIMP-1 but not MMP-2 in the HSCs.IL-10 inhibits TIMP-1 and MMP-2 expressions in HSCs induced by PDGF.
文摘目的:观察不同浓度葛根素对白细胞介素-1β(IL-1β)诱导的大鼠颈椎间盘纤维环细胞凋亡率及凋亡因子B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)基因与胱天蛋白酶-3(Caspase-3)的影响。方法:对大鼠颈椎间盘纤维环细胞进行体外培养3代,鉴定成功后分为5组:对照组,模型组,葛根素低、中、高浓度组。除对照组外,其余4组均给予10ng/m L IL-1β诱导细胞凋亡,同时葛根素低、中、高浓度组分别予以125、250、500μmol/L的葛根素进行干预,24h后用流式细胞仪检测各组纤维环细胞凋亡率,Western blot检测凋亡调控基因Caspase-3和Bcl-2蛋白表达。结果:葛根素能抑制IL-1β诱导的椎间盘纤维环细胞凋亡。Caspase-3蛋白表达水平比较:与对照组比较,模型组显著升高(P<0.05);与模型组比较,3个给药组均显著降低(P<0.01,P<0.05),且随着葛根素浓度的提高表达减弱,葛根素高浓度组与葛根素低、中浓度组比较差异均有统计学意义(P<0.05)。3个给药组Bcl-2蛋白表达水平均显著高于对照组和模型组,差异具有统计学意义(P<0.01)。结论:一定浓度葛根素能够有效地抑制大鼠颈椎间盘纤维环细胞的凋亡,其途径可能是通过上调抗凋亡因子Bcl-2,下调促凋亡因子Caspase-3蛋白的表达。
