Sun Meng, Yu Qin-Wei, Xiang Ting, Jiang Zhen-Zhou, Zhang Lu-Yong
Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University Nanjing 210009, China.
Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University Nanjing 210009, China Center for Drug Research and Development, Guangdong Pharmaceutical University Guangzhou 510006, China.
Zhongguo Zhong Yao Za Zhi. 2021 Jan;46(1):139-145. doi: 10.19540/j.cnki.cjcmm.20200818.401.
Polygonum multiflorum is a traditional Chinese herbal medicine and has many biological activities such as hair-blacking, anti-atherosclerosis, anti-inflammatory and anti-aging. However, the liver injury induced by P. multiflorum has aroused wide attention in recent years. 2,3,5,4'-tetrahydroxystibane-2-O-β-D-glucoside(TSG) is a main component of P. multiflorum, but the role of TSG in inducing liver injury is unclear. The aim of present study was to evaluate TSG's potential liver injury and effects on bile acid homeostasis and phospholipids efflux. C57 BL/6 J mice received intraperitoneal administration of 400 mg·kg(-1) of TSG daily for 15 days, and then biochemical indexes of liver injury and changes of phospholipid content were detected. The changes of bile acid compositions were detected by LC-MS/MS. The results showed TSG 400 mg·kg(-1) significantly increased the content of serum total bile acid(TBA) and alkaline phosphatase(ALP). Elevated free bile acid levels were observed in TSG-treated groups, including β-muricholic acid(β-MCA), ursodeoxycholic acid(UDCA), hyodeoxycholic acid(HDCA), chenodeoxycholic acid(CDCA), deoxcholic acid(DCA) in serum and β-MCA, CDCA in liver. TSG inhibited the protein expression of farnesoid X receptor(FXR) and down stream bile salt export pump(BSEP), which may result in the accumulation of bile acid. TSG also inhibited the expression of 25-hydroxycholesterol-7 alpha-hydroxylase(CYP7 B1), which may disturb the alternative pathway for bile acid synthesis. In addition, intraperitoneal injection of TSG 400 mg·kg(-1) significantly decreased the content of phospholipids in bile. The research showed that TSG significantly inhibited the expression of multidrug resistance protein 2(MDR2) and destroyed the regular distribution of MDR2 on the bile duct membrane of liver. In vitro results showed that the IC_(50) of TSG on HepG2 cells was about 1 500 μmol·L(-1) and TSG at 500 μmol·L~(-1)(for 24 h) could destroy the distribution of MDR2 on the bile duct membrane of liver. In conclusion, TSG induced liver injury by disrupting bile acid homeostasis and phospholipids efflux.
何首乌是一种传统的中药材，具有多种生物学活性，如乌发、抗动脉粥样硬化、抗炎和抗衰老等。然而，近年来何首乌引起的肝损伤已引起广泛关注。2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（TSG）是何首乌的主要成分，但TSG在诱导肝损伤中的作用尚不清楚。本研究的目的是评估TSG潜在的肝损伤以及对胆汁酸稳态和磷脂外排的影响。C57 BL/6 J小鼠每天腹腔注射400 mg·kg⁻¹的TSG，连续15天，然后检测肝损伤的生化指标和磷脂含量的变化。通过液相色谱-串联质谱法检测胆汁酸组成的变化。结果显示，400 mg·kg⁻¹的TSG显著增加了血清总胆汁酸（TBA）和碱性磷酸酶（ALP）的含量。在TSG处理组中观察到游离胆汁酸水平升高，包括血清中的β-鼠胆酸（β-MCA）、熊去氧胆酸（UDCA）、猪去氧胆酸（HDCA）、鹅去氧胆酸（CDCA）、脱氧胆酸（DCA）以及肝脏中的β-MCA、CDCA。TSG抑制了法尼酯X受体（FXR）和下游胆盐输出泵（BSEP）的蛋白表达，这可能导致胆汁酸的积累。TSG还抑制了25-羟基胆固醇-7α-羟化酶（CYP7 B1）的表达，这可能扰乱胆汁酸合成的替代途径。此外，腹腔注射400 mg·kg⁻¹的TSG显著降低了胆汁中磷脂的含量。研究表明，TSG显著抑制多药耐药蛋白2（MDR2）的表达，并破坏了MDR2在肝胆管膜上的正常分布。体外实验结果显示，TSG对HepG2细胞的半数抑制浓度（IC₅₀）约为1500 μmol·L⁻¹，500 μmol·L⁻¹的TSG（作用24小时）可破坏MDR2在肝胆管膜上的分布。综上所述，TSG通过扰乱胆汁酸稳态和磷脂外排诱导肝损伤。
