Licraside激活FXR缓解胆汁淤积的药理学研究  

Pharmacological research of Licraside in activating FXR to relieve cholestasis

作　　者：王国旭 则巴努尔·约麦尔江 张慧雨 席莉莉 张帆[2,4] 魏玉辉 WANG Guo-xu;ZEBANUER·Yue-mai-er-jiang;ZHANG Hui-yu;XI Li-li;ZHANG Fan;WEI Yu-hui(School of Pharmacy,Lanzhou University,Lanzhou 730000,Gansu Province,China;The First School of Clinical Medicine,Lanzhou University,Lanzhou 730000,Gansu Province,China;Office of Institution of Drug Clinical Trial,The First Hospital of Lanzhou University,Lanzhou 730000,Gansu Province,China;Departmen of Pharmacy,The First Hospital of Lanzhou University,Lanzhou 730000,Gansu Province,China)

机构地区：[1]兰州大学药学院,甘肃兰州730000 [2]兰州大学第一临床医学院,甘肃兰州730000 [3]兰州大学第一医院国家药物临床试验机构办公室,甘肃兰州730000 [4]兰州大学第一医院药剂科,甘肃兰州730000

出　　处：《中国临床药理学杂志》2024年第12期1759-1763,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81960646,82004080);兰州市科技局科技计划基金资助项目(2022-3-43)。

摘　　要：目的 研究异甘草素-4′-O-芹糖(1→2)葡萄糖苷(licraside)干预缓解胆汁淤积的分子作用机制。方法 用慢病毒沉默人肝癌HepG2细胞中法尼醇X受体(FXR),记为siFXR-HepG2细胞。将HepG2细胞和siFXR-HepG2细胞均分为正常组、模型组和实验组。模型组和实验组均给予胆红素+丙磺舒诱导高胆酸盐高胆红素模型;然后,正常组和模型组均不进行任何处理,实验组给予80μmol·L^(-1)licraside干预24 h。考察各组细胞的细胞活力及总胆汁酸(TBA)、胆红素等生化指标的含量,用蛋白质印迹法考察各组细胞FXR、胆酸盐输出泵(BSEP)等蛋白的表达水平。结果 HepG2细胞中正常组、模型组和实验组及siFXR-HepG2细胞中正常组、模型组和实验组的细胞活力分别为(100.00±17.15)%、(39.41±2.91)%、(70.79±3.74)%、(81.41±5.12)%、(33.49±2.69)%和(44.08±4.82)%,TBA水平分别为(7.98±5.87)、(46.18±10.93)、(9.25±7.20)、(11.18±3.36)、(38.28±5.12)和(34.79±5.39)μmol·L^(-1),总胆红素水平分别为(5.21±3.27)、(40.29±24.88)、(5.21±2.64)、(12.00±4.64)、(56.36±14.85)和(15.39±5.56)μmol·L^(-1),FXR蛋白相对表达水平分别为1.00±0.10、0.81±0.07、1.11±0.09、0.10±0.02、0.12±0.02和0.10±0.04,BSEP蛋白相对表达水平分别为1.00±0.17、0.81±0.02、0.88±0.03、0.70±0.09、0.49±0.07和0.60±0.10。HepG2细胞中实验组的上述指标和模型组相比,在统计学上差异均有统计学意义(P<0.001,P<0.01,P<0.05);除TBA外,siFXR-HepG2细胞中实验组的上述指标和模型组相比,在统计学上差异均有统计学意义(P<0.001,P<0.01)。结论 Licraside可以有效降低高胆酸盐高胆红素细胞的生化指标水平,这种作用是通过激动FXR的蛋白表达后,增加胆酸盐外排,减少胆汁酸的合成,达到缓解胆汁淤积的作用。Objective To investigate the molecular mechanism of action of isoglycoside-4'-O-apigenin(1→2) glucoside(licraside)intervention in alleviating cholestasis.Methods Farnesol X receptor(FXR) was silenced in human HepG2 cells using lentivirus,and bilirubin and α-isothiocyanate(ANIT) were used to induce HepG2cells to construct a hypercholate-hyperbilirubin model.The effects of licraside on cell viability,biochemical indices contents and the expression level of FXR and its downstream related proteins in the model were investigated.HepG2 cells and FXR-silenced HepG2 cells were divided into normal,model and experimental groups.Bilirubin and probenecid were added to all groups except the normal group,and 80 μmol·L^(-1) licraside was added to the experimental group.After 24 h of culture,cell viability and the levels of total bile acids(TBA),bilirubin and other biochemical indices were examined in each group;the protein expression levels of FXR and bile salt efflux pump(BSEP) were examined in each group by Western blot assay.Results The cell viability in HepG2 cells normal group,HepG2 cells model group,HepG2 cells experimental group,siFXR-HepG2 cells normal group,siFXR-HepG2 cells model group and siFXR-HepG2 cells experimental group were(100.00±17.15)%,(39.41±2.91)%,(70.79±3.74)%,(81.41±5.12)%,(33.49±2.69)% and(44.08±4.82)%;the levels of TBA were(7.98±5.87),(46.18±10.93),(9.25±7.20),(11.18±3.36),(38.28±5.12) and(34.79±5.39) μmol·L^(-1);the levels of total bilirubin were(5.21±3.27),(40.29±24.88),(5.21±2.64),(12.00±4.64),(56.36±14.85) and(15.39±5.56) μmol·L^(-1);the relative expression levels of FXR protein were 1.00±0.10,0.81±0.07,1.11±0.09,0.10±0.02,0.12±0.02 and 0.10±0.04;the relative expression levels of BSEP protein were 1.00±0.17,0.81±0.02,0.88±0.03,0.70±0.09,0.49±0.07 and 0.60±0.10.The differences of the above indexes in the HepG2 cells experimental group compared with the model group were statistically significant(P <0.001,P <0.01);except for TBA,the differences of the above index

关 键 词：异甘草素-4′-O-芹糖(1→2)葡萄糖苷(licraside) 法尼醇X受体 激动药 胆汁淤积 

分 类 号：R97[医药卫生—药品]