丁酸梭菌对肝缺血再灌注损伤大鼠的潜在保护作用

Potential Protective Effects of Clostridium Butyricum on Hepatic Ischemia Reperfusion Injury in Rats

目的:采用代谢组学方法研究丁酸梭菌预处理肝缺血再灌注损伤大鼠代谢物的变化和可能涉及的代谢途径。方法:将24只SD大鼠随机分为3组:假手术、模型组、益生菌组(术前5 d灌胃丁酸梭菌,2.5×10^(8)CFU·kg^(-1)·d^(-1)),每组8只。模型组和益生菌组大鼠构建肝脏缺血再灌注损伤模型。检测大鼠血清中ALT和AST的活性水平。利用超高效液相色谱-三重四级杆-线性离子阱质谱(UPLC-QTRAP-MS/MS)采集大鼠代谢物信息,并采用主成分分析法、正交偏最小二乘法判别分析法筛选差异代谢物,并做代谢通路分析。结果:与模型组相比,丁酸梭菌预处理后筛选出谷氨酸、谷氨酰胺、甘油磷胆碱、牛磺酸、乳酸、肌醇、琥珀酸盐等35种差异代谢物,主要涉及谷氨酰胺和谷氨酸代谢通路、牛磺酸和牛磺酸代谢通路等。与模型组相比,丁酸梭菌干预可显著降低肝缺血再灌注损伤大鼠血清ALT、AST活性(P<0.05)。结论:代谢组学方法结果表明丁酸梭菌能够调节大鼠缺血再灌注后的代谢平衡,从而起到保护肝缺血再灌注损伤大鼠的作用,这为今后研究丁酸梭菌治疗肝缺血再灌注损伤奠定了基础。

Objective: To investigate the changes of metabolites and metabolic pathways in rats with hepatic ischemia reperfusion injury pretreated with clostridium butyricum by metabonomic methods. Methods: Totally 24 SD rats were randomly divided into 3 groups: sham operation, model group and probiotic group(gavage clostridium butyricum 5 days before surgery, 2.5×10^(8)CFU·kg^(-1)·d^(-1)) with 8 rats in each group. The rats in the model group and probiotic group were used to construct liver ischemia-reperfusion injury models. The rat serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) activity levels were tested. The metabolites were collected by UPLC-QTRAP-MS/MS, the differential metabolites were screened by principal component analysis and orthogonal partial least square discriminant analysis, and the metabolic pathway was analyzed. Results: Compared with the model group, 35 differential metabolites such as glutamic acid, glutamine, glycerophosphocholine, taurine, lactate, myo-inositoland and succinate were screened out after pretreatment with clostridium butyricum. It was mainly related to glutamine and glutamate metabolic pathway, taurine and taurine metabolic pathway. Compared with the model group, clostridium butyricum intervention reduced the activities of ALT and AST in serum in hepatic ischemia reperfusion injury rats(P<0.05).Conclusion: The results of metabolomic methods show that clostridium butyricum can regulate the metabolic balance after ischemia reperfusion in rats, so as to protect hepatic ischemia reperfusion injury. This establishes a substantial foundation for future research and treatment.