脓毒症大鼠炎症与凝血相关性及药物干预的实验研究

Correlation between inflammation and blood coagulation in septic rats and experimental studies of drug intervention

目的 应用基因芯片技术探讨脓毒症大鼠炎症与凝血相关性及药物干预的影响,以期从基因水平上研究药物对其的作用机制.方法将90只Wistar大鼠随机分为对照组、模型组、药物组,每组30只.采用盲肠结扎穿孔术(CLP)复制脓毒症大鼠模型.药物组制模后腹腔注射泰能0.1 g/kg、血必净注射液10 ml/kg、连翘注射液10 ml/kg;对照组和模型组腹腔给予10 ml/kg生理盐水.72 h处死大鼠,用BiostarR-40 s芯片检测模型组/对照组、药物组/模型组大鼠肝组织基因,并以Cy3和Cy5荧光信号相对强度比值>2.0或<0.5筛选差异显著基因,通过美国国立生物技术信息中心(NCBI)数据库查询基因功能并加以分类.结果模型组/对照组筛选出差异基因305条,其中上调基因159条,下调基因146条;药物组/模型组筛选出差异基因342条,其中上调基因102条,下调基因240条;模型组、药物组已知功能共同差异表达基因36条,上调基因15条,下调基因21条;其中涉及免疫、代谢、物质转运、细胞凋亡等多种相关基因.结论药物组对脓毒症时发生紊乱的相关基因进行调控,使异常表达基因发生回归,特别是免疫相关基因及代谢相关基因中的超氧化物歧化酶2(SOD2)基因,使脓毒症病情得到良好的转归.

Objective To apply gene chip technique to explore the correlation between inflammation and blood coagulation of rats with sepsis and the effect of drug intervention to study the drug mechanism at gene level. Methods Ninety Wistar rats were randomly divided into three groups; control, model and drug groups (each n=30). The septic rat models were established by cecum ligation perforation (CLP) method. The drug group was treated by intraperitoneal injection of imipenem/cilastatin (0. 1 g/kg), Xuebijing injection (血必净注射液，10 ml/kg) and forsythia injection(连翘注射液，10 ml/kg) after CLP; while in the control and model groups, equal amount of normal saline was injected vie the same route. BiostarR-40，chip was used to detect the rat liver gene of model group/control group and drug group/model group, the ratio of fluorescein signal Cy3/Cy5 >2.0 or <0.5 was applied to screen the institias difference gene, through the database of American National Center of Biological Information <NCBI)，the gene function and classification were inquired. Results Model group/control group had 305 institias difference gene, including up-regulation gene 159 institias, and down-regulation gene 146 institias; drug group/model group had 342 institias difference gene, containing up-regulation gene 102 institias, and down-regulation gene 240 institias. In the model group and drug group, there were 36 genes with known functions and differential expressions, including up-regulation gene 15 inatitias and down-regulation gene 21 institias and involving immune, metabolism, material transportation and apoptosis related genes, and so on. Conclusion In the drug group, the drugs can regulate the occurrence of disturbances of related genes in rats with sepsis and promote the regression of gene abnormal expressions to take place, especially the superoxide dismutase-2 (SOD2) gene in the immune and metabolism related genes. Thus, the septic condition will have a good prognosis.