乌司他丁对脓毒症大鼠心脏组织基因表达的影响

Effect of ulinastatin preconditioning on gene expression profile of heart tissue in a rat sepsis model

目的 应用基因芯片技术观察乌司他丁（UTI）预处理对脓毒症大鼠心脏组织基因表达的调控作用.方法 45只雄性Wistar大鼠按照随机数字表法均分为对照组、脓毒症组和UTI组.采用盲肠结扎穿孔术（CLP）复制脓毒症大鼠模型;对照组仅开腹、关腹,不行CLP.UTI组制模前1 h肌肉注射（肌注）UTI100 kU/kg;脓毒症组及对照组肌注平衡液5 ml/kg.采用RatRef-12大鼠表达谱基因芯片进行检测,以Cy3和Cy5两种荧光信号强度结果比值＞2.0或＜0.5的基因为差异表达基因,用计算机软件筛选并分析比较脓毒症组和UTI组与对照组大鼠心脏组织基因表达的变化,并初步分析脓毒症组和UTI组表达基因之间的差异.结果 在22 523条基因中,与对照组比较,脓毒症组差异表达基因418条,占基因芯片总点数的1.856%,其中表达上调200条,已知功能基因84条,只在脓毒症组表达上调而UTI组表达正常者43条;表达下调218条,已知功能基因74条,只在脓毒症组表达下调而UTI组表达正常者37条.与对照组比较,UTI组共筛选出差异表达基因202条,占基因芯片总点数的0.897%,其中表达上调111条,已知功能基因57条,只在UTI组表达上调而脓毒症组表达正常者17条;表达下调91条,已知功能基因48条,只在UTI组表达下调而脓毒症组表达正常者18条.与对照组比较,UTI组和脓毒症组大鼠心脏组织基因表达同时上调41条,下调37条.结论 UTI预处理可减轻脓毒症晚期大鼠心脏的损害,具有一定的心脏保护效应;其基因机制可能涉及UTI对应激反应、细胞信号转导、物质能量代谢、免疫反应等方面相关基因表达的调控.

Objective To observe the regulatory effect of ulinastatin （UTI）preconditioning on gene expression of heart tissue in septic rats by DNA microarrays.Methods Forty-five male Wistar rats were equally divided into control group, sepsis group and UTI group by means of random number table.Cecal ligation and puncture （CLP） was used to reproduce rat sepsis model.The control group only experienced a simulated operation without CLP.In UTI group the rats were treated with intramuscular injection of UTI 100 kU/kg 1 hour before CLP.In sepsis group and control group balanced electrolyte solution （5 ml/kg） was given.Gene expression spectrum was studied with RatRef-12 rat gene expression profile microarray to detect the changes in gene expression pattern of rat heart tissue after CLP.Genes with fluorescent signal of Cy3/Cy5 of ratio average （RA）＞2.0 or RA＜0.5 were identified as differential genes,and those highly correlated to sepsis and UTI groups were screened by means of related computer software to analyze their relationship.Results In 22 523 genes,418 differential genes were found in sepsis group compared with control group,accounting for 1.856%, and among them 200 genes showed up-regulation, with 84 known functional genes,and 43 of which only showed up-regulation in sepsis group,but normal in UTI group.Two hundred and eighteen genes showed down-regulation, with 74 known functional genes, 37 of which only showed down-regulation in sepsis group, but normal in UTI group.Two hundred and two differential genes were found in UTI group compared with control group, accounting for 0.897%, and among them 111 genes showed up-regulation, with 57 known functional genes, and 17 of which only showed up-regulation in UTI group, but normal in sepsis group.Ninety-one genes showed down-regulation, with 48 known functional genes, 18 of which only showed down-regulation in UTI group, but normal in sepsis group.Compared with the control group, in both UTI group and sepsis group, 41 of known functional genes showed up-regulation,and 37 showed down-regulation.Conclusion UTI preconditioning can ameliorate the damage to heart tissue in rat sepsis model, thus it has a protective effect on heart, and its mechanism may be attributable to regulatory effect of UTI on expression of stress reaction, cell signal transduction, energy metabolism,immune reaction and other related genes.