Analysis of Transport Mechanism of Cyperotundone in Caco-2 Cell Model

Hui-ling GUO; Qiang HU; Lyu-jiang HU; Wen-jun GAO; Zhi-fang HU; Xiao-juan ZHAO

doi:10.13422/j.cnki.syfjx.20191547

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Analysis of Transport Mechanism of Cyperotundone in Caco-2 Cell Model

Pharmacy and Processing | 更新时间：2021-02-09

- Analysis of Transport Mechanism of Cyperotundone in Caco-2 Cell Model
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 23, Pages: 110-115(2019)
- 作者机构：
  
  1.江西中医药大学，南昌　 330004；
  2.江西中医药高等专科学校，江西　抚州　 344000
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191547
  CLC： R22; Q2; R28; C37; R945; O657
- Received：29 December 2018，
  
  Published Online：15 April 2019，
  
  Published：05 December 2019
- 稿件说明：
移动端阅览
Hui-ling GUO, Qiang HU, Lyu-jiang HU, et al. Analysis of Transport Mechanism of Cyperotundone in Caco-2 Cell Model[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(23): 110-115.
DOI：

Hui-ling GUO, Qiang HU, Lyu-jiang HU, et al. Analysis of Transport Mechanism of Cyperotundone in Caco-2 Cell Model[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(23): 110-115. DOI： 10.13422/j.cnki.syfjx.20191547.

摘要

目的：

考察香附烯酮在Caco-2细胞模型中的转运机制，为香附的临床应用提供实验依据。

方法：

通过噻唑蓝(MTT)比色法考察不同质量浓度香附烯酮对Caco-2细胞的毒性，以确定转运试验的给药浓度。以香附烯酮表观渗透系数(

app

)和累积转运量为评价指标，采用液相色谱-质谱联用技术(LC-MS)测定香附烯酮的含量，色谱条件为流动相乙腈(A)-水(B)梯度洗脱(0～1.5 min，35%A；1.5～2 min，35%～90%A；2～4 min，90%A；4～4.1 min，90%～35%A；4.1～8 min，35%A)，流速0.3 mL·min

－1

，进样量1 μL，柱温30℃；质谱条件为电喷雾离子源(ESI)，正离子模式，香附烯酮及内标物蛇床子素的检测离子分别为

219.2～110.9，

245.0～189.0。探讨了香附烯酮质量浓度、给药时间、乙二胺四乙酸(EDTA)及P-糖蛋白(P-gp)抑制剂对香附烯酮在体外细胞模型上跨膜转运的影响。

结果：

香附烯酮质量浓度处于3～90 mg·L

－1

时，孵育4 h内对Caco-2细胞没有明显毒性。香附烯酮在Caco-2细胞模型中的转运存在一定的浓度及时间依赖性，其

app

1×10

－6

cm·s

－1

，说明药物吸收良好，外排率处于0.5～1.5；加入细胞旁路转运抑制剂EDTA及P-gp转运体抑制剂维拉帕米后，双向

app

没有显著性差异。

结论：

香附烯酮在Caco-2细胞模型转运机制主要是被动扩散，细胞旁路转运及P-gp不参与其转运。

Abstract

Objective:

To investigate transport mechanism of cyperotundone in Caco-2 cell model and provide experimental basis for clinical application of Cyperi Rhizoma.

Method:

The toxicity of cyperotundone with different concentrations to Caco-2 cells was investigated by methyl thiazolyl tetrazolium (MTT) colorimetry

in order to determine the concentration of administration in transport test. The content of cyperotundone was determined by liquid chromatography-mass spectrometry (LC-MS) with apparent permeability coefficient (

app

) and cumulative transport capacity as indexes. The chromatographic conditions were as following: mobile phase of acetonitrile (A)-water (B) for gradient elution (0-1.5 min

35%A; 1.5-2 min

35%-90%A; 2-4 min

90%A; 4-4.1

90%-35%A; 4.1-8 min

35%A)

the flow rate at 0.3 mL·min

-1

injection volume of 1 μL

and temperature of column at 30 ℃. The mass spectrometric conditions was electrospray ionization (ESI) and positive ion mode

the detection ions of cyperotundone and osthole (internal standard substance) were

219.2-110.9 and

245.0-189.0

respectively. Effect of concentration of cyperotundone

administration time

ethylenediamine tetraacetic acid (EDTA) and P-glycoprotein (P-gp) inhibitor on the transmembrane transport of cyperotundone on

in vitro

cell model were investigated.

Result:

Cyperotundone didn't have significant toxicity to Caco-2 cells at 3-90 mg·L

-1

after incubation for 4 h. The transportion of cyperotundone in Caco-2 cell model was related to the concentration and time to a certain extent

its

app

was higher than 1×10

-6

cm·s

-1

which indicated that absorption of cyperotundone was good

the efflux rate (ER) of cyperotundone was 0.5-1.5.There was no significant difference in bidirectional

app

of cyperotundone after the addition of cell bypass transport inhibitor (EDTA) and P-gp transport inhibitor (verapamil).

Conclusion:

The transport mechanism of cyperotundone in Caco-2 cell model is mainly passive diffusion

and cell bypass transport and P-gp are not involved in its transport.

关键词

Keywords

references

Artursson P , Palm K , Luthman K . Caco-2 monolayers in experimental and theoretical predictions of drug transport [J]. Adv Drug Deliv Rev , 2001 , 46 ( 1/3 ): 27 - 43 .

慈倩倩，李英霞，张文，等．香附的本草考证 [J]．四川中医， 2010 ， 28 ( 4 )： 50 - 51 ．

LIN X S , WU H Q , HUANG F . Analysis of essential oils from Cypcrus rotundus by GC/MS [J]. J Chin Mass Spectrometr Soc , 2006 , 27 ( 1 ): 40 - 44 .

FENG Y F , GUO X L , MENG Q , et al . Study on the chemical substrates of SFE extract from Rhizoma Cypcri [J]. J Chin Med Mater , 2006 , 29 ( 3 ): 232 - 235 .

郭慧玲，董能峰，胡律江，等．基于成分敲出策略辨识四制香附抗痛经的主要效应成分 [J]．中国实验方剂学杂志， 2017 ， 23 ( 10 )： 7 - 11 ．

郭慧玲，骆云霞，胡律江，等．四制香附贴膏灸剂在大鼠体内的血液流变性与体外透皮特性分析 [J]．中国实验方剂学杂志， 2017 ， 23 ( 11 )： 29 - 33 ．

胡律江，许茜茜，赵晓娟，等．混料均匀设计优化四制香附活性成分配伍 [J]．中国实验方剂学杂志， 2015 ， 21 ( 14 )： 1 - 3 ．

胡律江，郭慧玲，胡志方，等． LC-MS考察α-香附酮在Caco-2细胞模型中转运特性 [J]．中国实验方剂学杂志， 2014 ， 20 ( 5 )： 35 - 38 ．

李雪，李翠，崔学军，等． HPLC-MS/MS测定人血浆中青藤碱浓度及芪麝丸在人体内的药代动力学分析 [J]．中国实验方剂学杂志， 2017 ， 23 ( 9 )： 85 - 89 ．

陈腾飞，刘建勋，姚明江，等．三七皂苷R1脑、血液微透析探针的体内外回收率分析 [J]．中国实验方剂学杂志， 2017 ， 23 ( 15 )： 35 - 40 ．

Kigen G , Edwards G . Drug-transporter mediated interactions between anthelminthic and antiretroviral drugs across the Caco-2 cell monolayers [J]. BMC Pharmacol Toxicol , 2017 , 18 ( 1 ): 20 .

廖沙，谢剑炜． Caco-2细胞模型在药物体外研究中的应用 [J]．中国新药杂志， 2005 ， 14 ( 4 )： 416 - 419 ．

孔令提，刘冬，宋春丽，等．采用Caco-2细胞转运模型研究胺碘酮转运机制及阿托伐他汀对其转运的影响 [J]．中国医院用药评价与分析， 2016 ， 16 ( 11 )： 1472 - 1474 ．

杨秀伟，杨晓达，王莹，等．中药化学成分肠吸收研究中Caco-2细胞模型和标准操作规程的建立 [J]．中西医结合学报， 2007 ， 5 ( 6 )： 634 - 641 ．

高坤，孙进，何仲贵． Caco-2细胞模型在口服药物吸收研究中的应用 [J]．沈阳药科大学学报， 2005 ， 22 ( 6 )： 73 - 78 ．

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