创伤弧菌脓毒症大鼠肝脏基质金属蛋白酶及其抑制基因的表达
Gene expression of matrix metalloprotease and their inhibitors in liver tissue of rats with Vibrio vulnificus sepsis
摘要目的 观察创伤弧菌脓毒症大鼠肝脏组织基质金属蛋白酶(MMP-2、MMP-9)及其抑制基因(TIMP-2、RECK)表达水平的变化,探讨它们在创伤弧菌脓毒症中的作用.方法 清洁级Sprague-Dawley雄性大鼠60只,随机分为6组,每组10只.第1组为健康对照组,第2~6组为创伤弧菌脓毒症模型组.观察大鼠行为学改变,提取各组大鼠肝脏组织总RNA,半定量RT-PCR法检测MMP-2、MMP-9以及TIMP-2、RECK基因mRNA水平表达的变化.结果 染菌4 h后,大鼠出现呼吸急促,皮温升高,下肢肿胀,且症状进行性加重;染菌12 h后,大鼠出现紫绀、间断抽搐等症状.染菌2、6和9 h后,大鼠肝脏组织中MMP-2及MMP-9基因mRNA水平表达明显上调,TIMP-2和RECK表达水平下调.结论 MMP-2和MMP-9表达上调,以及TIMP-2和RECK表达水平下调提示它们可能参与了创伤弧菌脓毒症的病理生理过程.
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abstractsObjective To investigate gene expressions of matrix metalloprotease (MMP-2, MMP-9) and tissue inhibitor of metalloproteinase-2 (TIMP-2) and RECK ( reversion inducing cysteine-rich protein with Kazal motifs) in the liver tissue of rats with Vibrio vulnificus sepsis. Methods Vibrio vulnificus sepsis model was induced by injection of 1 mL/100 g Vibrio vulnificus (9 × 106 cfu/mL) at hind limbs in 50 Sprague-Dawley male rats of clean grade, each time 10 animals were sacrificed at 2, 6, 9, 12 and 16 h after injection and liver samples were taken. Ten rats served as control group. The ethological changes were observed. The total RNA was extracted from liver tissue and the gene expressions of MMP-2, MMP-9, TIMP-2 and RECK were evaluated by semi-quantitative RT-PCR. Results The rats manifested shortness of breath, fever and swelling in hind limbs at 4 h after injection of Vibrio vulnificus. These symptoms gradually deteriorated, and the rats presented cyanosis and convulsion at 12 h. The gene expressions of MMP-2 and MMP-9 were markedly up-regulated, while those of TIMP-2 and RECK were down-regulated. Conclusion The study suggests that the elevation of MMP-2 and MMP-9 and down-regulation of TIMP-2 and RECK is associated with the development of Vibrio vulnificus sepsis.
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