姜黄素对亚急性肝内胆汁淤积大鼠肠内菌群变化的影响及其机制研究

目的： 研究姜黄素对亚急性肝内胆汁淤积（intrahepatic cholestasis，IHC）大鼠肠内菌群变化的影响，并分析相关机制。方法： 38只IHC大鼠随机分为模型组（9只）、姜黄素低剂量组（9只）、姜黄素高剂量组（10只）、阳性药物组（10只），9只健康大鼠设置为对照组。姜黄素低、高剂量组姜黄素200、400 mg·kg^-1灌胃；阳性药物组熊去氧胆酸60 mg·kg^-1灌胃；对照组与疾病组等量生理盐水灌胃。均为每天一次；干预14 d。检测肝功能指标、胆红素代谢指标、炎症指标；qRT-PCR反应定量检测肠内菌群；Western blot检测肝组织核转录因子-κB（NF-κB）p65、p-NF-κB p65、核因子抑制蛋白（IκBα）、p-IκBα蛋白相对表达量。结果： 与疾病组比较，姜黄素低、高剂量组、阳性药物组碱性磷酸酶（ALP）、丙氨酸氨基转移酶（ALT）、谷草转氨酶（AST）、γ-谷氨酰转肽酶（GGT）、总胆红素（TBIL）、直接胆红素（DBIL）、总胆汁酸（TBA）、白介素-1β（IL-1β）、白介素-6（IL-6）、转化生长因子-β₁（TGF-β₁）均降低，白介素-10（IL-10）均升高，乳酸杆菌、双歧杆菌均增加，大肠杆菌均减少（P<0.05）；与姜黄素低剂量组比较，姜黄素高剂量组、阳性药物组ALP、ALT、AST、GGT、TBIL、DBIL、TBA、IL-1β、IL-6、TGF-β₁均降低，IL-10均升高，乳酸杆菌、双歧杆菌均增加，大肠杆菌均减少（P<0.05）。与疾病组比较，姜黄素低、高剂量组、阳性药物组p-NF-κB p65/NF-κB p65，p-IκBα/IκBα均降低（P<0.05）；与姜黄素低剂量组比较，姜黄素高剂量组、阳性药物组p-NF-κB p65/NF-κB p65，p-IκBα/IκBα均降低（P<0.05）。结论： 姜黄素可改善亚急性IHC模型大鼠肝功能、胆红素代谢、肠道菌群，减轻炎症反应及肝脏组织病理变化，且具有剂量依赖性，推测其作用机制与抑制NF-κB信号通路有关。

Effect of curcumin on the changes of intestinal flora in rats with subacute intrahepatic cholestasis and its mechanism

OBJECTIVE To explore the effect of curcumin on the changes of intestinal flora in rats with subacute intrahepatic cholestasis (IHC) and elucidate the related mechanism.METHODS A total of 38 IHC rats were randomly divided into four groups of model group (n=9), curcumin low-dose (n=9), curcumin high-dose (n=10) and positive drug (n=10). Nine healthy rats were designated as control group. The curcumin low/high-dose group received an intragastric dose of curcumin 200/400 mg·kg^-1 while the positive drug group ursodeoxycholic acid 60 mg·kg^-1. Both control and disease groups received one daily intragastric dose of the same amount of normal saline for 14 days. The parameters of liver function, bilirubin metabolism and inflammation were detected. And quantitative real-time polymerase chain reaction (qRT-PCR) was employed for quantifying intestinal flora. HE stain was used for observing the pathological changes of liver tissue and Western blot for detecting the relative expressions of p-NF-κB p65, NF-κB p65, p-IκBα and IκBα protein in liver tissue.RESULTS As compared with disease group, alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), total bilirubin (TBIL), direct bilirubin (DBIL), total bile acid (TBA), interleukin-1β (IL-1β), interleukin-6 (IL-6) and transforming growth factor-β₁ (TGF-β₁) declined in curcumin low/high-dose and positive drug groups. Interleukin-10 (IL-10), Lactobacillus and Bifidobacterium rose while Escherichia coli decclined (P<0.05). As compared with curcumin low-dose group, ALP, ALT, AST, GGT, TBIL, DBIL, TBA, IL-1β, IL-6, and TGF-β₁ declined in curcumin high-dose and positive drug groups. IL-10, Lactobacillus and Bifidobacterium rose while Escherichia coli declined (P<0.05). As compared with disease group, p-NF-κB p65/NF-κB p65 and p-IκBα/IκBα declined in curcumin low/high-dose and positive drug groups (P<0.05). As compared with curcumin low-dose group, p-NF-κB p65/NF-κB p65 and p-IκBα/IκBα declined in curcumin high-dose and positive drug groups (P<0.05).CONCLUSION In a dose-dependent fashion, curcumin can improve liver function, bilirubin metabolism and intestinal flora, reduce inflammation and liver tissue pathological changes in subacute IHC model rats. And its mechanism of action may be correlated with an inhibition of NF-κB signaling pathway.