四君子汤治疗酒精性肝损伤的效果及其机制

EFFICACY OF SI-JUN-ZI DECOCTION IN THE TREATMENT OF ALCOHOLIC LIVER DISEASE AND ITS MECHANISM

目的探讨四君子汤治疗酒精性肝损伤的效果及其机制。方法将雄性C57BL/6小鼠随机分为正常组(A组)、酒精模型组(B组)、四君子汤低剂量组(C组)及四君子汤高剂量组(D组),B~D组小鼠每天灌胃体积分数0.56的乙醇溶液(6 g/kg),A组小鼠每天灌胃等剂量的生理盐水;同时C、D组小鼠再分别每天灌胃5、10 g/kg的四君子汤,每5 d测1次小鼠体质量,实验持续18 d。灌胃结束后,采集小鼠新鲜粪便进行16S rDNA基因测序分析小鼠肠道菌群的变化;麻醉后,眼眦取血,收集血清进行肠道通透性检测;收集小鼠血液和肝脏、脾脏组织样本,测定各组小鼠的肝、脾指数以及血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(T-BIL)、碱性磷酸酶(ALP)、三酰甘油(TG)和总胆固醇(TC)的含量,利用苏木精-伊红(HE)和油红O染色观察各组小鼠肝、脾组织形态学变化。结果D组与B组相比,小鼠体质量及肝指数、脾指数差异有显著性(t=3.53~4.41,P<0.05),血清中ALP、T-BIL、TG水平显著降低(t=3.51~5.60,P<0.05);C、D组与B组相比,小鼠血清中ALT、AST、TC水平显著下降(t=2.19~7.62,P<0.05)。病理结果显示C、D组与B组比较,肝组织中肝索形态恢复正常,肝脏脂肪蓄积减轻;脾组织中红髓白髓分界清晰。肠道通透性结果显示,D组与B组比较,肠道通透性显著降低(t=3.19,P<0.05);肠道菌群检测结果显示,C、D组较B组,小鼠肠道菌群多样性增加,Dubosiella、Bifidobacterium、Allobaculum等有益菌含量增多,Prevotellaceae_UCG-001、Alloprevotella等致病菌含量降低。结论四君子汤可有效改善酒精诱导的小鼠肝损伤,其机制可能是通过调节肠道菌群变化实现的。

Objective To explore the efficacy of Si-jun-zi Decoction in treating alcoholic liver disease and its mechanism.Methods Male C57BL/6 mice were randomly divided into normal group(group A),alcohol model group(group B),low-dose Si-jun-zi Decoction group(group C),and high-dose Si-jun-zi Decoction group(group D).Mice in the groups B-D were intragastrically administered ethanol solution(6 g/kg)with a volume fraction of 0.56 per day,while mice in group A were intragastrically administered an equal dose of normal saline per day.Meanwhile,mice in groups C and D were given Si-jun-zi Decoction at 5 and 10 g/kg per day,respectively.The body mass was measured once every 5 d.The experiment lasted for 18 d.After gavage,fresh feces were collected to analyze the changes in intestinal flora by 16S rDNA gene sequencing.After anesthesia,blood was collected from the canthus,and serum was collected for intestinal permeability test.Blood,liver,and spleen tissues were collected.The liver and spleen indices as well as the content of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(T-BIL),alkaline phosphatase(ALP),triglyceride(TG),and total cholesterol(TC)were measured in each group.The morphological changes of liver and spleen were observed in each group using hematoxylin-eosin and oil red O staining.Results Compared with group B,group D had significantly different body mass and liver and spleen indices(t=3.53-4.41,P<0.05),and significantly lower serum ALP,T-BIL,and TG levels(t=3.51-5.60,P<0.05).Compared with group B,groups C and D had significantly decreased serum ALT,AST,and TC levels(t=2.19-7.62,P<0.05).The pathological results showed that compared with group B,the morphology of hepatic cords in the liver tissues of groups C and D were back to normal,and the accumulation of liver fat was reduced;the red marrow and white marrow in the spleen tissue were clearly demarcated.The results of intestinal permeability showed that the intestinal permeability of group D was significantly lower than that of group B(t=3.19,P<0.05).The results of intestinal flora showed that compared with group B,the diversity of intestinal flora in mice of groups C and D was increased,with the content of beneficial bacteria such as Dubosiella,Bifidobacterium,and Allobaculum increased and that of pathogenic bacteria such as Prevotellaceae_UCG-001 and Alloprevotella decreased.Conclusion Si-jun-zi Decoction effectively ameliorates alcohol-induced liver injury in mice.The mechanism may be achieved by regulating intestinal flora.