丙酮酸乙酯对脓毒症小鼠细胞因子信号转导调节机制研究

Research of Signal Transduction Mechanisms of Cytokines in Sepsis Treated with Ethyl Pyruvate

目的:探讨丙酮酸乙酯对小鼠脓毒症细胞因子信号转导调节机制。方法:选用雄性昆明小鼠60只,随机分成三组:A组:盲肠结扎穿孔(CLP)模型+丙酮酸乙酯林格式液(REPS)复苏组(n=20)。B组:CLP模型+林格氏液(RLS)复苏组(n=20)。C组:假手术组(n=20)。手术后24小时获取血清及肝脏标本。监测指标包括:应用双抗体夹心酶联免疫吸附测定法测定小鼠血清TNFα、IL-6、IL-10浓度。采用分光光度法测定肝脏组织NO和血清NO浓度。采用逆转录聚合酶链反应(reverse transcription-polymerasechain reaction,RT-PCR)方法测定肝组织iNOSmRNA、IL-6mRNA、TNFαmRNA表达。结果:CLP组TNFα浓度、IL-6浓度、IL-10浓度、肝组织和血清NO浓度较假手术组高,差异有统计学意义,P<0.01;LSD组间比较显示,REPS组TNFα、IL-6浓度较RLS组低,IL-10浓度REPS组较RLS组增高,肝组织和血清NO浓度REPS组较RLS组显著下降,差异有统计学意义,P<0.01。CLP组小鼠肝细胞TNFαmRNA、IL-6mRNA、iNOSmRNA表达量均较假手术组增加,差异有统计学意义,P<0.01;组间LSD检验提示,RLS组TNFαmRNA、IL-6mRNA、iNOSmRNA表达量较REPS组升高,差异有统计学意义,P<0.01。结论:EP通过调控脓毒症细胞因子基因表达,可以使体内促炎/抗炎反应达到一个相对稳定状态。

Objective： To investigate the signal transduction mechanisms of cytokines treated with ethyl pyruvate in septic mice. Methods 60 mice were randomly divided into three groups. Cecal ligation-perforation （CLP） model were made in mice as septic model. Ringer＇s ethyl pyruvate solution （REPS） and Ringer＇s lactic solution （RLS） were used to resuscitate septic mice. Mice were divided into three groups： A group： CLP ＋ REPS （n=20）; B group： CLP ＋ RLS （n=20）. C group： sham operation （n=20）. 24 hours after operation, mice were sacrificed to gain serum and hepatic tissue, nitric oxide （NO） in liver and serum NO were measured using chromatometry. Serum tumor necrosis factor alpha （TNFet）, interleukin 6 （IL-6） and interleukin 10 （IL-10） were measured with enzyme-linked im- munosorbentassay （ELISA）. Hepatocyte TNFctmRNA, IL-6 mRNA and inducible of nitric oxide synthase （iNOS） mRNA expression were measured with reverse transcriptase polymerase chain reaction （RT-PCR）. Results： Comparing with sham operation group, CLP mouse have a higher level of serum TNFo~, IL-6, IL-10 and NO （P〈0.01）. Analysised by Fisher＇s protected LSD test, REPS group has a lower level in serum TNFot, IL-6, NO （including liver and serum） and a higher level in serum IL-10, compared with RLS group （P〈0.01）. Comparing with sham operation group, CLP mouse have a higher level of hepatocyte TNFet mRNA, IL-6 mRNA and iNOS mRNA ex- pression （P〈0.01）. Analyzed by Fisher＇s protected LSD test, REPS group has a lower level ofhepatocyte TNFot mRNA, IL-6 mRNA and iNOS mRNA expression compared with RLS group （P〈0.0）. Conclusion： Ethyl pyruvate modulates the concentration of serum cytokines, and decreases the cytok^e gene expression in hepatic tissue which helps to achieve cytokine balance and ameliorate systemic inflammation.