摘要
目的:观察高压氧加自由基拮抗剂对大鼠肝脏损伤早期及肝纤维化形成过程中肝脏基质金属蛋白酶-2(MMP-2)表达的影响方法:用 CCl_4建立大鼠肝纤维化模型,在2wk 和8wk 分别给予高压氧结合自由基拮抗剂、高压氧、自由基拮抗剂处理,放免法测定血清透明质酸(HA)的含量,以了解细胞外基质合成情况,免疫组化法检测肝组织 MMP-2蛋白以及其调节相关的生长因子 TGF-β1 蛋白的表达;应用逆转录-聚合酶链反应(RT-PCR)方法检测 MMP-2 mRNA 水平.结果:实验2wk 末模型组高压氧结合自由基拮抗剂组肝脂肪变性和水样变性及点状坏死与模型组相比明显减轻,MMP-2蛋白及 mRNA,TGF-β1蛋白表达均低于模型组,差异显著(MMP-2:5.04±1.34 vs 56.75±3.03,P<0.01;MMP-2mRNA:0.155±0.005 vs 0.547±0.013.P<0.01;TGF-β1:3.28±0.33 vs 4.96±0.28,P<0.01).血清 HA虽低于模型组,但无显著差异(P>0.05).实验8wk 末模型组肝小叶内纤维结缔组织增生明显并形成粗细不等的纤维条索,高压氧结合自由基拮抗剂组以肝细胞脂变和细胞水肿为主,纤维结缔组织无明显增生,肝组织 MMP-2蛋白及 mRNA,TGF-β1蛋白表达和血清 HA 水平与模型组相比均明显为低(MMP-2:12.53±2.51 vs 160.57±5.54, P<0.01;MMP-2 mRNA:0.506±0.013 vs 0.685±0.014,P<0.01;TGF-β1:4.97±0.35 vs 7.76±0.39,P<0.01;HA:116.8±17.7μg/L vs 87.8±31.6μg/LP<0.01).高压氧组、自由基拮抗剂组在2wk 末和8wk 末肝损伤略轻于模型组,MMP-2蛋白和 TGF-β1蛋白表达均低于模型组,且高压氧组 MMP-2 mRNA 表达亦较模型组低.结论:高压氧结合自由基拮抗剂可以降低 CCl_4损伤大鼠肝脏 MMP-2表达,可以迟滞肝纤维化病变的发展.
AIM:To study the effects of hyperbaric oxygen combined with free radical antagonists(HFr)on the expression of matrix metalloproteinase-2(MMP-2)at the early stage of liver injury and fibrogenesis in rat. METHODS:A liver fibrosis model was induced by carbon tetrachloride(CCl_4)in Wistar rats,and the rats were ex- posed to hyperbaric oxygen,free radical antagonists and both of them combined together respectively during the 2nd and 8th week.Serum hylaluronate(HA)level was detected by radioimmunoassay to explore extra cellular matrix changes.The content of MMP-2 and TGF-β1,which was a kind of growth factor being able to regulate the expression of MMP-2,were examined by immunohis- tochemical staining.A semi-quantitative RT-PCR method was used to evaluate MMP-2 mRNA expression. RESULTS:The hepatocytic injury including fatty degener- ation,hydropic changes and spotty necrosis in HFr group was lighter than that in model group at the 2nd weekend, meanwhile,the MMP-2 protein expression and mRNA level, TGF-β1 protein expression of the liver tissues were lower than those in the model significantly(MMP-2:5.04±1.34 vs 56.75±3.03,P<0.01;MMP-2 mRNA:0.155±0.005 vs 0.547±0.013,P<0.01;TGF-β1:3.28±0.33 vs 4.96±0.28, P<0.01).The level of serum HA was also lower than that in the model,but not significantly.At the 8th weekend,in the model,fibrous connective tissue proliferated markedly in hepatic Iobules,forming fibrous septa.Fatty degeneration, hydropic changes were observed in HFr group,in which the expression of MMP-2 protein and its mRNA,TGF-β1 protein,serum HA were all lower significantly compared with the models(MMP-2:12.53±2.51 vs 160.57±5.54, P<0.01;MMP-2 mRNA:0.506±0.013 vs 0.685±0.014, P<0.01;TGF-β1:4.97±0.35 vs 7.76±0.39,P<0.01;HA: 116.8±17.7μg/L^(-1) vs 87.8±31.6μg/L^(-1)P<0.01).In addition, the hepatocytic lesions in hyperbaric oxygen group,free radical antagonists group were similar to that in model, but somewhat mild.MMP-2 and TGF-β1 proteins,MMP-2 mRNA expression in hyperbaric oxygen group were lowe
出处
《世界华人消化杂志》
CAS
2003年第12期1909-1914,共6页
World Chinese Journal of Digestology
