目的 确证芍药苷对α-异硫氰酸酯(α-isothiocyanate,ANIT)诱导胆汁淤积大鼠的药效作用,通过16SrRNA技术探究芍药苷对肠道菌群的调节作用机制。方法 SD大鼠随机分为对照组、模型组及芍药苷高、中、低剂量(200、100、50mg/kg)组和熊去氧胆酸(60mg/kg)组,每组10只。给予药物干预7d,第4天igANIT(60mg/kg)诱导胆汁淤积模型。测定胆汁体积流量;检测血清中血清中天冬氨酸氨基转移酶(aspartate aminotransferase,AST)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、γ-谷氨酰转肽酶(γ-glutamyl transpeptidase,γ-GT)活性及总胆红素(total bilirubin,TBIL)、直接胆红素(direct bilirubin,DBIL)、总胆汁酸(total bileacid,TBA)水平;采用苏木素-伊红(HE)染色观察肝脏组织病理变化;收集大鼠肠内容物进行肠道菌群测序,探索芍药苷对肠道菌群的影响。结果 与模型组比较,芍药苷有效改善胆汁体积流量(P<0.05),显著下调大鼠血清中肝功能指标AST、ALT、γ-GT活性及胆汁淤积指标TBIL、DBIL、TBA水平(P<0.01),改善肝细胞损伤。肠道菌群测序结果可知,芍药苷显著上调厚壁菌门与拟杆菌门比值及异杆菌属Allobaculum、布劳特氏菌属Blautia丰度(P<0.05),降低颤螺菌属Oscillospira、瘤胃球菌属Ruminococcus丰度(P<0.05、0.01)。结论 芍药苷可能通过改善ANIT诱导的胆汁淤积大鼠肠道菌群失调,从而发挥对胆汁淤积肝损伤的改善作用。

Objective To verify the pharmacodynamic effect of paeoniflorin on α-isothiocyanate (ANIT)-induced cholestasis in rats, and explore the regulatory mechanism of paeoniflorin on intestinal flora by 16S rRNA technology. Methods SD rats were randomly divided into control group, model group, paeoniflorin high-, medium-and low-dose (200, 100, 50 mg/kg) groups and ursodeoxycholic acid (60 mg/kg) group, with 10 rats in each group. After 7 d of drug intervention, model of cholestasis was induced by ig ANIT (60 mg/kg) on 4th day. Bile volume flow was measured; Activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (γ-GT) and levels of total bilirubin (TBIL), direct bilirubin (DBIL), total bile acid (TBA) in serum were detected; Hematoxylin eosin (HE) staining was used to observe the pathological changes of liver tissue; Intestinal contents of rats was collected for intestinal flora sequencing to explore the effect of paeoniflorin on intestinal flora. Results Compared with model group, paeoniflorin effectively improved bile volume flow (P< 0.05), significantly down-regulated liver function index AST, ALT, γ-GT activities and cholestatic index TBIL, DBIL and TBA levels (P< 0.01), improved the damage of liver cells. The sequencing results of intestinal flora showed that paeoniflorin significantly increased the ratio of Firmicutes/Bacteroides and abundance of Allobaculum and Blautia (P< 0.05), and decreased the abundance of Oscillospira and Ruminococcus (P< 0.05, 0.01). Conclusion Paeoniflorin may improve the liver injury of cholestasis by improving the imbalance of intestinal flora in ANIT-induced cholestasis rats.

R285.5

国家自然科学基金资助项目（82274187）；四川省科技厅应用基础项目（2019YJ0492）