Metabolism Analysis

Effect of anemoside B4 on expression and activity of CYP1A2*

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  • 1. Jiangxi University of Traditional Chinese Medicine,Nanchang 330006,China;
    2. State Key Laboratory of Innovative Drugs and Efficient Energy- saving Pharmaceutical Equipment,Nanchang 330006,China;
    3. Jiangxi Muentang Biological Technology Co.,Ltd.,Nanchang 330006,China

Received date: 2020-08-21

  Online published: 2024-07-15

Abstract

Objective: To study the effect of anemoside B4 on expression and activity of CYP1A2 in vivo and in vitro,to provide experimental data for possible interaction between aemoside B4 and other drugs. Methods: The chromatographic separation was carried out using a UHPLC XB-C18 column(2.1 mm×50 mm,1.8 μm)with mobile phase comprising of 0.1% fomic acid-acetonitrile at a flow rate of 0.3 mL·min-1,the quantitation was performed using multiple reaction monitoring of the transitions of m/z 180.1 → 109.9 in positive electrospray ionization. The liver micrisomes of normal mice was prepared. The incubation system was established. Liver micrisomes was incubated with probe phenacetin(4 μg·mL-1)and B4(50 μg·mL-1),UPLC-MS/MS was adopted to detect the content of phenacetin after incubation for 5,15,30,60 and 120 min. Incubation time and content were used to draw a linear equation,the a value in the equation reflecting the metabolic rate of phenacetin was analyzed. Mice were intravenous administrated with Anemoside B4 for 3 days,then the probe phenacetin (2 mg·kg-1)was injected intraperitoneally,plasma was separated for the detection of phenacetin(UPLC-MS/ MS)after 5,15,30 and 60 min of probe injection,the liver was removed for mRNA expression by qPCR assay. Protein expression was achieved with Western blot assay. Results: The linearity range was 4-2 000 ng·mL-1r2=0.993 6), the limit of detection was 0.1 ng·mL-1S/N ≥ 3)and limit of quantification was 0.2 ng·mL-1S/N ≥ 10),intraday and interday relative standard deviations were less than 5% and 10% respectively,the recoveries were between 88.5%- 92.9%. Anemoside B4 has no effect on metabolic rate of phenacetin in vitroP>0.05),but significantly promote the metabolic rate in plasma of mice(P<0.01)and upregulate the mRNA and protein expression of CYP1A2. Conclusions: In vitro,anemoside B4 has no effect on CYP1A2 activity,it has a mild stimulative effect on CYP1A2 in vivo,which is related to the up-regulation of gene transcription and protein expression of CYP1A2.

Cite this article

GONG Qin, WANG Mu-lan, LI Jun-mao, HUANG Wen-ping, FENG Yu-lin, FU Ai-dong, YANG Shi-lin, LI Jun . Effect of anemoside B4 on expression and activity of CYP1A2*[J]. Chinese Journal of Pharmaceutical Analysis, 2021 , 41(6) : 970 -978 . DOI: 10.16155/j.0254-1793.2021.06.05

References

[1] 缪成贵,周国梁,秦梅颂,等.白头翁皂苷对佐剂性关节炎大鼠FZD8 表达的影响[J].中国中药杂志,2015,40(20):4063
LIAO CG,ZHOU GL,QIN MS,et al.Effect of pulchinenoside on FZD8 expression of adjuvant arthritis rats[J].Chin J Chin Mat Med,2015,40(20):4063
[2] XU QM,SHU Z,HE WJ,et al.Antitumor activity of Pulsatilla chinensis(Bunge)Regel saponins in human liver tumor 7402 cells in vitro and in vivo[J].Phytomedicine,2012,19(3/4):293
[3] 陈凯,张文明,喻凯,等.白头翁中抗肿瘤物质的筛选及其作用机制研究[J].天然产物研究与开发,2016,28(12):1875.
CHEN K,ZHANG WM,YU K,et al.Screening of antitumor compounds from Radix Pulsatillae and researching on their mechanism[J].Nat Prod Res Dev,2016,28(12):1875
[4] 罗颖颖,陈兰英,简晖,等.白头翁皂苷调节 Bel-7402 人肝癌异体移植瘤裸鼠能量代谢的研究[J].中草药,2014,4(7):973 LUO YY,CHEN LY,JIAN H,et al.Regulation of saponins from Pulsatilla chinensis on energy metabolism of Bel-7402 xenograft in nude mice[J].Chin Trad Herb Drugs,2014,4(7):973
[5] 罗颖颖,陈兰英,崔亚茹,等.白头翁皂苷抑制人 HT29 结肠癌细胞增殖及诱导凋亡作用研究[J].中药药理与临床,2013,29(5):52
LUO YY,CHEN LY,CUI YR,et al.Effects of Pulsatilla chinensis saponins on proliferation and apoptosis of HT29 cells[J].Pharm Clin Chin Mat Med,2013,29(5):52
[6] LIU M,ZHAO X,XIAO L,et al.Cytotoxicity of the compounds isolated from Pulsatilla chinensis saponins and apoptosis induced by 23-hydroxybetulinic acid[J].Pharm Biol,2015,53(1):1
[7] CHEN Z,GUAN Y,ZHOU L,et al.Preparation and characterization of colon-targeted particles of Pulsatilla chinensis Saponins[J].Nat Prod Communic,2015,10(2):237
[8] 赵光伟,王玉舜,蔡乐,等.中药白头翁对猪传染性胸膜肺炎的辅助疗效[J].当代畜牧,2015,325(29):97
ZHAO GW,WANG YS,CAI L,et al.The auxiliary effect of Pulsatilla on infectious pleural pneumonia in pigs[J].Contemp Anim Husbandry,2015,325(29):97
[9] 缪成贵,周国梁,秦梅颂,等.白头翁皂抑制佐剂性关节炎大鼠成纤维样滑膜细胞凋亡[J].中南大学学报(医学版),2015,40(2):144
LIAO CG,ZHOU GL,QIN MS,et al.Pulchinenoside inhibits the fibroblast-like synoviocytes apoptosis in adjuvant arthritis rats[J]. J Cen South Univ(Med Sci),2015,40(2):144
[10] 段强军,云云,陈志珍,等.白头翁汤正丁醇提取物对小鼠外阴阴道念珠菌病的治疗作用研究[J].中国真菌学杂志,2017,12(6):337
DUAN QJ,YUN Y,CHEN ZZ,et al.Butyl alcohol extract of Baitouweng decoction inhibit vulvovaginal candidiasis in murine model[J].Chin J Mycol,2017,12(6):337
[11] 胡唐玲,施高翔,徐志庆,等.白头翁汤正丁醇提取物对白念珠菌细胞壁抑制作用研究[J].中国真菌学杂志,2018,13(5):262 HU TL,SHI GX,XU ZQ,et al.Inhibitory effect of butyl alcohol extract of Baitouweng decoction on candidaalbicanscell wall[J]. Chin J Mycol,2018,13(5):262
[12] 王淑英,刘萌萌,吴银萍,等.白头翁提取物体外抗结核杆菌作用的实验研究[J].江西中医学院学报,2009,21(5):42
WANG SY,LIU MM,WU YP,et al.Experimental study of effects of Chinese bulbul extracts againsting mycobacterium tuberculosis in vitro[J].J Jiangxi Univ Tradit Chin Med,2009,21(5):42
[13] 司红彬,吴永继,徐素萍,等.白头翁水提物与抗菌药联合对耐药大肠埃希菌作用研究[J].动物医学进展,2015,36(11):49
SI HB,WU YJ,XU SP,et al.Study on effect of Pulsatilla chinensis water extract combined with antimicrobials against drug-resistant E. coli[J].Prog Vete Med,2015,36(11):49
[14] 王晓妹,田歌,段强军,等.白头翁汤正丁醇提取物对白念珠菌定植下溃疡性结肠炎小鼠的作用机制研究[J].中国中药杂志, 2018,43(14):2979
WANG XM,TIAN G,DUAN DQ,et al.Therapeutic potential of n-butanol extract of Pulsatilla decoction in a murine model of ulcerative colitis induced by DSS combined with Candida albicans colonization[J].Chin J Chin Mate Med,2018,43(14):2979
[15] 李俊,左沙沙,邱小萱,等.炎性和凋亡因子在小鼠急性肾损伤中的表达特点及白头翁皂苷 B4 的调节作用[J].中国中药杂志, 2020,45(1):157
LI J,ZUO SS,QIU XX,et al.Expression characteristics of inflammatory and apoptosis factors and regulatory effect of anemoside B4 in mice with acute kidney injury[J].Chin J Chin Mate Med, 2020,45(1):157
[16] 龚琴,王木兰,左沙沙,等.结扎大鼠肾动静脉所致急性肾损伤模型的研究[J].中国中药杂志,2019,44(5):996
GONG Q,WANG ML,ZUO SS,et al.Study on acute kidney injury model induced by renal ischemia-reperfusion in rats[J].Chin J Chin Mater Med,2019,44(5):996
[17] 龚琴,王木兰,何鹿玲,等.白头翁皂苷 B4 对实验性急性肾功能损伤的保护作用[J].中国现代中药,2019,21(1):62
GONG Q,WANG ML,HE LL,et al.Protective effect of anemoside b4 from experimental acute renal damage in animal[J].Mod Chin Med,2019,21(1):62
[18] 李俊,左沙沙,何鹿玲,等.白头翁皂苷 B4 对结扎大鼠肾动静脉所致缺血再灌注损伤的治疗作用及相关机制研究[J].中国中药杂志,2020,45(3):617
LI J,ZUO SS,HE LL,et al.Study on therapeutic effect and its related mechanism of anemoside B4 on ischemia reperfusion injury by renal artery and vein ligation in rats[J].Chin J Chin Mat Med, 2020,45(3):617
[19] 梁淼,马增春,易剑峰,等.四物汤及其配伍对大鼠肝脏 P450 酶活性及 mRNA 表达的影响[J].中国中药杂志,2013,38(21): 3720
LIANG M,MA ZC,YI JF,et al.Effect of Siwu decoction and its combined administration on hepatic P450 enzymatic activity and mRNA expression in rats[J].Chin J Chin Mate Med,2013,38(21):3720
[20] 李晓宇,刘皋林.CYP450 酶特性及其应用研究进展[J].中国临床药理学与治疗学,2008,13(8):942
LI XY,LIU HL.Research advance on the characteristics and application of CYP450 metabolic enzymes[J].Chin J Clin Pharmacol Ther,2008,13(8):942
[21] 常彬霞,貌盼勇,辛绍杰.Ⅰ相药物代谢酶细胞色素 P450 及其与肝脏疾病的关系[J].解放军医学杂志,2010,35(3):333
CHANG BX,MAO PY,XIN SJ.Phase Ⅰ drug metabolism enzymes cytochrome P450 and its relationship with liver disease[J].Med J Chin PLA,2010,35(3):333
[22] 姜敬非,王玉刚,胡珺,等.新药用资源菠萝叶对小鼠相关药物代谢酶的诱导作用[J].世界科学技术,2012,14(6):2149
JIANG JF,WANG YG,HU J,et al.CYPs induction of polyphenol extract from new medicinal resource Ananas Comosus leaf on mice[J].World Sci Tech,2012,14(6):2149
[23] 苏克,刘付丽,温从丛,等.超高效液相色谱-串联质谱仪同时检测大鼠血浆中 5 种 CYP450 探针药及其应用[J].中国卫生检验杂志,2018,28(16):1947
SU K,LIU FL,WEN CC,et al.Simultaneous determination of bupropion,metroprolol,midazolam,phenacetin and tolbutamide in rat plasma by UPLC-MS/MS[J].Chin J Health Lab Techhol, 2018,28(16):1947
[24] GONG Q,YAN XJ,LEI F,et al.Proteomic profiling of the neurons in mice with depressive-like behavior induced bycorticosterone and the regulation of berberine:pivotal sites of oxidative phosphorylation[J].Mol Brain,2019,12:118
[25] GONG Q,HE LL,WANG ML,et al.Comparison of the TLR4/NFκB and NLRP3 signalling pathways in major organs of the mouse after intravenous injection of lipopolysaccharide[J].Pharm Biol, 2019,57(1):555
[26] 成碟,徐为人,刘昌孝.细胞色素P450(CYP450)遗传多态性研究进展[J].中国药理学通报,2006,22(12):1409
CHENG D,XU WR,LIU CX.Progress of research on genetic polymorph ism of CYP450 s[J].Chin Pharm Bull,2006,22(12): 1409
[27] 叶娜,万丽,杨秋楠,等.丹参注射液对人肝微粒体酶 CYP2C9, CYP2C19,CYP2D6 体外抑制作用的研究[J].药物评价研究, 2014,37(6):502
YE N,WAN L,YANG QN,et al.In vitro inhibition of Danshen injection on human liver cytochrome P450 enzyme CYP2C9, CYP2C19,CYP2D6[J].Drug Eval Res,2014,37(6):502
[28] 肖鹏,周宏灏.细胞色素氧化酶 CYP1A2 的研究进展[J].中南大学学报(医学版),2008,33(5):456
XIAO P,ZHOU HH.Advance in CYP 1A2[J].J Central South Univ(Med Sci),2008,33(5):456
[29] 袁靖,赵军宁,李祖伦.肝药代谢酶研究方法及中药影响的研究进展[J].四川中医,2007,25(6):33
YUAN J,ZHAO JN,LI ZL.The research progress of liver drug metabolizing enzymes and the influence of traditional Chinese medicine[J].J Sichuan Tradit Chin Med,2007,25(6):33
[30] 辛勤,梁国威,刘会臣.CPY1A2 基因多态性对药物代谢的影响[J].国际检验医学杂志,2013,34(24):3378
XIN Q,LI ANG GW,LI U HC.Effect of CPY1A2 gene polymorphism on drug metabolism[J].Int J Lab Med,2013, 34(24):3378
[31] 鲍珍贝,杨青,胡敏,等.西洋参总皂苷对大鼠肝微粒体 CYP1A2活性的影响[J].实用药物与临床,2017,20(3):250
BAO ZB,YANG Q,HU M,et al.Influence of total saponins from Panax quinquefolium Linne on the activity of CYP1A2 in rats[J]. Pract Pharm Clin Rem,2017,20(3):250
[32] 张艳辉,于超,郭延垒,等.高效液相色谱法测定鼠肝微粒体中CYP1A2 酶的活性及动力学考察[J].药物分析杂志,2012,32(2):189
ZHANG YH,YU C,GUO YL.HPLC determination ofactivity of CYP1A2 and its pharmacokineticsin rat liver microsomes[J].Chin J Pharm Anal,2012,32(2):189
[33] 洪世忠,何帆,陈卫东.左归饮对大鼠体内 Cyp3a1、Cyp2d2、 Cyp1a2 代谢酶活性的研究[J].贵阳中医学院学报,2018,40(4):17
HONG SZ,HE F,CHEN WD.A Study on the effect of Zuoguiyin on metabolic enzymes activity of Cyp3a1,Cyp2d2 and Cyp1a2 in rats[J].J Guiyang Univ Chin Med,2018,40(4):17
[34] BERESFORD AP,ELLIS WJ,AYRTON J,et al.Cytochrome P4501A(CYP1A)induction in rats and man by the benzodioxino derivative,fluparoxan[J].Xenobiotica,1997,27(2):159
[35] ZHU Q,LIAO J,XIE L,et al.Mechanism-based inhibition of CYP1A2 by antofloxacin,an 8-NTH-derivative of levofloxacin in rats[J].Xenobiotica,2009,39(4):293
[36] 张轶雯,钟里科,楼倩雯,等.川芎嗪诱导小鼠肝药酶 Cyp3a11 的作用及其机制研究[J].中国现代应用药学,2018,35(7):967
ZHANG YW,ZHONG LK,LUO QW,et al.Effect and mechanism of ligustrazine induced liver drug metabolism enzyme Cyp3a11[J]. Chin J Mod Appl Pharm,2018,35(7):967
[37] 徐利云,侯丛颂,杨志宏,等.LC-MS/MS 法研究 Cocktail 探针药物咖啡因、右美沙芬、奥美拉唑、咪达唑仑及其代谢产物在大鼠血浆中的药代动力学[J].中国药理学通报,2017,33(2):268
XU LY,HOU CS,YANG ZH,et al.Pharmacokinetics studies on caffeine,dextromethorphan,omeprazole,midazolam and their metabolites in rat plasma by LC-MS/MS[J].Chin Pharm Bull, 2017,33(2):268
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