College of Pharmaceutical Sciences, Key Laboratory of Luminescence Analysis and Molecular Sensing (Ministry of Education), Southwest University, No. 2 Tiansheng Road, Chongqing 400715, P.R. China.
School of Life Sciences, Southwest University, No. 2 Tiansheng Road, Chongqing 400715, P.R. China.
Phytomedicine. 2021 Jul;87:153588. doi: 10.1016/j.phymed.2021.153588. Epub 2021 Jun 3.
Cholestasis is characterized by accumulation of bile components in liver and systemic circulation. Restoration of bile acid homeostasis via activating farnesoid x receptor (FXR) is a promising strategy for the treatment of cholestasis. FXR-SHP (small heterodimer partner) axis plays an important role in maintaining bile acid homeostasis.
To investigate the anti-cholestasis effect of Dolomiaea souliei (Franch.) C.Shih (D. souliei) and clarify its underlying mechanism against α-naphthylisothiocyanate (ANIT) induced acute intrahepatic cholestasis.
ANIT-induced Sprague-Dawley rats were employed to investigate the anti-cholestasis effect of D. souliei ethyl acetate extract (DSE). Ursodeoxycholic acid (UDCA) was used as positive control. Bile flow and blood biochemical parameters were measured. Liver histopathological examination was conducted via hematoxylin-eosin staining. Western blot analysis was carried out to evaluate the protein levels related to bile acids metabolism and inflammation. The interactions between FXR and costunolide or dehydrocostus lactone, were conducted by molecular docking experiments. The effect of costunolide and dehydrocostus lactone on aspartate aminotransferase (AST), alanine aminotransferase (ALT) levels and FXR expression were also evaluated using guggulsterone-induced L02 cells.
DSE could promote bile excretions and protect against ANIT-induced liver damage in cholestasis rats. Protein levels of FXR, SHP, Na/taurocholate cotransporter (NTCP), bile salt export pump (BSEP), multidrug resistance-associated protein 2 (MRP2) were increased and the expressions of cholesterol 7α-hydroxylase (CYP7A1) and sterol 27-hydroxylase (CYP27A1) were decreased by DSE. Meanwhile, the anti-inflammatory factors, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) were also significantly increased, and the pro-inflammatory factor, interleukin-10 (IL-10), was significantly decreased in rats of DSE groups. Molecular docking revealed that costunolide and dehydrocostus lactone could be well docked into the FXR protein molecule, and hydrophobic interactions played the main function. Costunolide could reverse the increased AST and ALT levels and increase the FXR expression in guggulsterone-induced L02 cells.
DSE had an anti-cholestasis effect by activating FXR-SHP axis, inhibiting synthesis of bile acid, and increasing bile secretion, together with inflammatory response and improving liver injury. Costunolide may be the main active component. This study provided a potential therapeutic mechanism for D. souliei as an anti-cholestasis medicine in the treatment of cholestasis liver diseases.
胆汁淤积症的特征是胆汁成分在肝脏和全身循环中积聚。通过激活法尼醇 X 受体 (FXR) 恢复胆汁酸动态平衡是治疗胆汁淤积症的一种有前途的策略。FXR-SHP（小异二聚体伴侣）轴在维持胆汁酸动态平衡中起着重要作用。
研究紫菀（Dolomiaea souliei (Franch.) C.Shih）对急性肝内胆汁淤积的抗胆汁淤积作用，并阐明其对α-萘异硫氰酸酯 (ANIT) 诱导的急性肝内胆汁淤积的作用机制。
采用 ANIT 诱导的 Sprague-Dawley 大鼠研究紫菀乙酸乙酯提取物 (DSE) 的抗胆汁淤积作用。熊去氧胆酸 (UDCA) 用作阳性对照。测量胆汁流量和血液生化参数。通过苏木精-伊红染色进行肝组织病理学检查。通过 Western blot 分析评估与胆汁酸代谢和炎症相关的蛋白水平。通过分子对接实验研究了法尼醇和去氢木香内酯与 FXR 的相互作用。还使用 guggulsterone 诱导的 L02 细胞评估了木香内酯和去氢木香内酯对天冬氨酸氨基转移酶 (AST)、丙氨酸氨基转移酶 (ALT) 水平和 FXR 表达的影响。
DSE 可促进胆汁排泄并防止胆汁淤积大鼠的 ANIT 诱导的肝损伤。FXR、SHP、Na/牛磺胆酸钠共转运蛋白 (NTCP)、胆汁盐输出泵 (BSEP)、多药耐药相关蛋白 2 (MRP2) 的蛋白水平升高，胆固醇 7α-羟化酶 (CYP7A1) 和固醇 27-羟化酶 (CYP27A1) 的表达降低。同时，抗炎因子肿瘤坏死因子-α (TNF-α)、白细胞介素-1β (IL-1β)、白细胞介素-6 (IL-6) 也显著增加，抗炎因子白细胞介素-10 (IL-10) 也显著减少。分子对接表明，木香内酯和去氢木香内酯可与 FXR 蛋白分子很好地对接，并且疏水相互作用起主要作用。木香内酯可逆转 guggulsterone 诱导的 L02 细胞中 AST 和 ALT 水平的升高，并增加 FXR 的表达。
DSE 通过激活 FXR-SHP 轴、抑制胆汁酸合成、增加胆汁分泌、炎症反应和改善肝损伤，发挥抗胆汁淤积作用。木香内酯可能是主要的活性成分。该研究为紫菀作为治疗胆汁淤积性肝病的抗胆汁淤积药物提供了潜在的治疗机制。
