清瘟败毒饮对脓毒症大鼠脾组织TNF信号通路相关基因表达的影响

Effect of Qing wen Baidu Decoction on the expression of TNF signal pathway related genes in the spleen of septic rats

目的基于TNF信号通路探讨清瘟败毒饮抗脓毒性脾功能损伤的可能机制。方法选择清洁级健康雄性Wistar大鼠45只,随机分为对照组、脓毒症组(A组)、清瘟败毒饮干预组(B组),每组15只。A组大鼠采用盲肠结扎穿孔术(CLP)进行复制脓毒症模型;B组大鼠除CLP外,于术前2天给予中药胃饲2次/d,术后继续中药灌胃;对照组实施除CLP外的开腹、关腹。3组术毕均予5 ml/kg平衡液肌肉注射。术后24 h取脾组织提取总RNA后行RNA-seq对mRNA表达量进行检测,并应用生物信息学方法分析清瘟败毒饮对脓毒症大鼠脾组织TNF信号通路相关基因的表达变化。结果相对于对照组,A、B组大鼠脾组织TNF信号通路相关基因中,同时上调的基因有19个,同时下调的基因有8个,下调转正常的基因1个,正常转上调的基因5个,由上调转为正常的基因7个(Ccl20、Cxcl10、IL-1b、IL6、Bcl3、Mmp9、Sele),正常转为下调的基因13个(ASK1、IKKα、CASP8、ITCH、p38、Drp1、IKKs、p38、Ccl5、Csf1、IL18R1、IL15、Ifi47)。结论清瘟败毒饮可通过调节TNF信号通路相关基因的过度表达,从而对脓毒症大鼠脾功能起到保护作用。

Objective To explore the possible mechanism of Qing wen Baidu Decoction(QWBD)against the damage of spleen function due to sepsis based on TNF signal pathway.Methods Forty five clean male Wistar rats were randomly divided into control group,sepsis group(A group)and Qing wen Baidu Decoction intervention group(B group).Rats in group A were treated with cecal ligation and perforation(CLP)to establish sepsis model.Besides CLP,rats in group B were fed with Chinese medicine twice a day two days before operation.After operation,the same dosage of traditional Chinese medicine was continue.In the control group,the abdomen was just opened and closed beside CLP.All rats of the three groups were given intramuscular injection by 5ml/kg balance solution.24 hours after operation,the total RNA were extracted from the spleen and the RNAseq was used to detect the mRNA expression.The expressions of TNF signal pathway related genes in the spleen of septic rats were analyzed by bioinformatics.Results Compared with the control group,group A and group B,there were 19 genes of up-regulation,8 genes of down-regulation in both groups,1 gene from down-regulation to normal expression,5 genes from normal expression to up-regulation,7 genes from up-regulation to normal(CCL20,CXCL10,IL-1B,IL6,BCl3,MMP9,sele),and 13 genes from normal expression to down-regulation:(ASK1,IKKα,CASP8,ITCH,p38,Drp1,IKKs,p38,Ccl5,Csf1,IL18R1,IL15,Ifi47).Conclusion Qing wen Baidu Decoction may protect spleen function of sepsis rats by regulating the over expression of TNF signal pathway related genes.