PK11195对hCAR亚型发挥拮抗作用的分子机制探讨  

Explore the antagonistic mechanisms of PK11195 against hCAR subtypes

作　　者：沈阗阗 石阿茜 夏文彬 蒋珍秀[2] 魏玉辉[1] 姚小军[5] 席莉莉[3] SHEN Tian-tian;SHI A-xi;XIA Wen-bin;JIANG Zhen-xiu;WEI Yu-hui;YAO Xiao-jun;XI Li-li(Department of Pharmacy,The First School of Clinical Medicine,Lanzhou University/The First Hospital of Lanzhou University,Lanzhou 730000,Gansu Province,China;Department of Neurology,The First School of Clinical Medicine,Lanzhou University/The First Hospital of Lanzhou University,Lanzhou 730000,Gansu Province,China;Office of Institution of Drug Clinical Trial,The First School of Clinical Medicine,Lanzhou University/The First Hospital of Lanzhou University,Lanzhou 730000,Gansu Province,China;Department of Pharmacy,Lanzhou University,Lanzhou 730000,Gansu Province,China;State Key Laboratory of Quality Research in Chinese Medicine,Macao University of Science and Technology,Macao 999078,China)

机构地区：[1]兰州大学第一临床医学院兰州大学第一医院药剂科,甘肃兰州730000 [2]兰州大学第一临床医学院兰州大学第一医院神经内科,甘肃兰州730000 [3]兰州大学第一临床医学院兰州大学第一医院药物临床试验机构办公室,甘肃兰州730000 [4]兰州大学药学院,甘肃兰州730000 [5]澳门科技大学中药质量研究国家重点实验室,中国澳门999078

出　　处：《中国临床药理学杂志》2022年第10期1083-1087,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81960646);甘肃省自然科学基金资助项目(20JR5RA370);甘肃省高等学校产业支撑计划基金资助项目(2020C-02);甘肃省卫生行业科研计划基金资助项目(GSWSKY-2019-84);兰州大学第一医院院内基金资助项目(ldyyyn2018-30);甘肃省教育厅优秀研究生“创新之星”基金资助项目(2021CXZX-142)。

摘　　要：目的以PK11195作为工具分子,探究其分别对hCAR不同亚型(hCAR1/2/3)拮抗的作用机制。方法用荧光素酶报告基因实验检测了不同浓度的PK11195干预后,对hCAR1、hCAR2和hCAR3的拮抗活性,随后用分子对接方法从原子水平上探讨了PK11195与hCAR1/2/3的相互作用模式,测量hCAR1-PK11195、hCAR2-PK11195和hCAR3-PK11195复合物结构配体结合域中每个对应残基的主链原子间的距离。结果荧光素酶报告基因实验结果显示:与空白组相比,5μmol·L^(-1) PK11195可以使hCAR1的基础活性显著降低(1.01±0.06 vs 0.48±0.06,P<0.001),但对hCAR2和hCAR3的基础活性降低并无显著性差异。分子对接结果显示,PK11195与hCAR1/2/3对接打分值分别为-9.73,-7.47和-7.74,将hCAR1-PK11195复合物分别与hCAR2-PK11195、hCAR3-PK11195复合物的三维结构叠合,原子间距离的均方根偏差值分别为3.87和0.22?,发现在helix 11、helix X和helix 12结构区域均具有明显的结构差异。结论PK11195对hCAR亚型活性具有不同的调节作用,其诱导的拮抗作用是通过共抑制因子结合而失活(helix 4-helix 11接触)。Objective To investigate the antagonistic mechanism of PK11195 as a tool molecule against hCAR1/2/3.Methods The antagonistic activity of PK11195 against different subtypes of human CAR(hCAR1,hCAR2 and hCAR3)was detected by luciferase reporter gene assay.Subsequently,the interaction mode between PK11195 and hCAR1/2/3 was explored at the atomic level by molecular docking method.The distances between main chain atoms of each corresponding residue in the ligand binding domain of hCAR1-PK11195,hCAR2-PK11195 and hCAR3-PK11195 complex structure were measured.Results The results of luciferase reporter gene assay showed that PK11195 significantly reduced the basal activity of hCAR1 when the intervention dose was 5μmol·L^(-1) compared with the control group(1.01±0.06 vs 0.48±0.06,P<0.001),but there was no significant difference in the basal activity of hCAR2 and hCAR3;PK11195 was docked with hCAR1/CAR2/CAR3,with scores of-9.73,-7.47 and-7.74.hCAR1-PK11195 protein was superimposed with hCAR2-PK11195 complex and hCAR3-PK11195 complex,respectively.The root mean square deviation(RMSD)values of interatomic were 3.87 and 0.22?,and there were significant structural differences in helix 11,helix X and helix 12 regions.Conclusion PK11195 has different regulatory effects on hCAR subtypes,the antagonism induced by PK11195 was inactivated by co-inhibitory factor binding(helix 4-helix 11 contact).

关 键 词：组成型雄甾烷受体 亚型 荧光素酶报告基因 分子对接 

分 类 号：R97[医药卫生—药品]