酸枣仁汤对慢性睡眠剥夺小鼠肝功能和特定肠道菌的影响

Effect of Suan-Zao-Ren Decoction on liver function and specific gut microbiota in mice with chronic sleep deprivation

目的从肠道菌群角度探讨酸枣仁汤对慢性睡眠剥夺小鼠肝功能的影响。方法C57BL/6雄性小鼠随机分为对照组、模型组和酸枣仁汤组。采用强迫运动睡眠剥夺法建立14 d的慢性睡眠剥夺模型,酸枣仁汤组在睡眠剥夺同时给予酸枣仁汤,对照组正常饲养,不作处理。观察造模前后小鼠体质量和摄食量变化;收集粪便样本,提取粪便基因组DNA,实时荧光定量PCR(qRT-PCR)检测特定肠道菌的差异;同时收集血清样本,试剂盒检测丙氨酸转氨酶(ALT)和总胆红素(TBIL)的活性,进而指示各组肝功能的变化。结果与对照组比较,模型组小鼠的摄食量显著增加(P<0.05),体质量显著下降(P<0.05),给予酸枣仁汤显著增加睡眠剥夺小鼠的进食量(P<0.05)。与对照组比较,睡眠剥夺小鼠的TBIL结果未见明显差异;而模型组ALT明显升高(P<0.05),同时给予酸枣仁汤可逆转ALT的升高(P<0.05)。qRT-PCR结果显示,模型组小鼠粪便中大肠杆菌属和产气荚膜梭菌与对照组比较显著增加(P<0.05),而双歧杆菌属、罗氏菌属、乳酸菌属与对照组比较显著降低(P<0.05)。同时给药可以逆转特定肠道菌大肠杆菌属、产气荚膜梭菌和乳酸菌属的变化(P<0.05),对双歧杆菌属、罗氏菌属没有显著影响。睡眠剥夺小鼠粪便拟杆菌门/厚壁菌门比有下降趋势,酸枣仁汤组该趋势升高,没有显著性差异。结论酸枣仁汤可能通过改善肠道菌群,缓解慢性睡眠剥夺导致的肝损伤。

Objective To investigate the effect of Suan-Zao-Ren Decoction(SZRD)on liver function in mice with chronic sleep deprivation from the perspective of gut microbiota.Methods C57BL/6 mice were randomly divided into three groups:control group,model group,SZRD group.A 14-day chronic sleep deprivation model was established by using the forced exercise sleep deprivation method.SZRD was given once a day during the 14-day sleep deprivation.The changes of body weight and food intake of mice before and after modeling was observed.Fresh fecal samples were collected for each mouse at the end of the experiment.Fecal genomic DNA was extracted to analyze the gut microbiota by real-time quantitative PCR(qRT-PCR).Serum samples were collected,and the activity of ALT and TBIL indicating changes in liver function were detected.Results The average food intake of all sleep-deprived mice was significantly increased(P<0.05),while the weight has no obvious increase(P<0.05)compared to the control mice.SZRD treatment further increased the food intake of sleep-deprived mice(P<0.05).Compared with control group,there was no significant difference in TBIL activity in sleep-deprived mice.ALT activity was significantly increased in model group(P<0.05),and SZRD treatment reversed the increase in ALT activity(P<0.05).qPCR results showed that E.coli and Clostridium perfringens in the feces of model group were significantly increased compared with the control group(P<0.05),while Bifidobacteria,Roseburia and Lactobacillus were significantly decreased(P<0.05).Meanwhile,SZRD treatment reversed the changes of E.coli,Clostridium perfringens and Lactobacillus(P<0.05),but had no effect on the changes of Bifidobacteria and Roseburia.The ratio of Bacteroidetes to Firmicutes in the feces of sleep-deprived mice decreased,SZRD treatment increased it.And the changes had no statistical significance.Conclusion SZRD can alleviate liver injury caused by chronic sleep deprivation by improving gut microbiota.