中国临床药理学杂志2024，Vol.40Issue(9)：1272-1276,5.DOI:10.13699/j.cnki.1001-6821.2024.09.007
基于3D肝细胞球模型评价何首乌醇提物毒性研究
Toxicity evaluation of alcohol extract of Polygonum multiflorum based on 3D hepatocyte ball model
摘要
Abstract
Objective To explore the toxicity of Polygonum multiflorum alcohol extract on 3D hepatospheres.Methods Variations in culture conditions and cell ratios were implemented,followed by the assessment of cell sphere diameter,density,and roundness,aiming to explore the optimal culture conditions.The 3D hepatocyte spheres were divided into control group and experimental-L,-M,-H groups.The experimental-L,-M,-H groups were treated with 0.25,1.00 and 2.50 mg·mL-1 Polygounm multiforum alcohol extract,and the control group was given the same amount of culture medium.The cell viability of the cell spheroids was tested by CellTiter-Glo reagent,the expression level of liver function related genes was detected by fluorescent quantitative polymerase chain reaction(RT-qRCR).The toxicity of cell spheres was detected by double fluorescent staining of living and dead cells.Results The ideal culture condition of cell sphere was 500 cells per micropore,and the cell ratio was HepG2-Huvec-LX-2=8∶1∶1.It displayed the values of 0.91±0.07 for circularity,0.91±0.02 for firmness,1.12±0.14 for aspect ratio,and(170.97±14.79)μm for diameter.On the 3rd,7th,10th and 14th days,the expression levels of albumin(ALB)mRNA were 1.00±0.02,0.96±0.02,0.54±0.07,0.52±0.07,and the expression levels of cytochrome P450 1A2(CYP1A2)mRNA were 1.00±0.10,2.15±0.16,2.45±0.33,1.30±0.03,respectively.The expression levels of multidrug resistance protein 2(MPR2)in the control group and the experimental-L,-M,-H groups were 1.00±0.31,1.38±0.24,1.48±0.06 and 1.90±0.08,respectively;spheroid viability were(98.19±0.49)％,(88.53±0.90)％,(71.60±2.91)％and(56.65±5.41)％.There were statistically significant differences in the above indexes between the experimental-L,-M,-H groups and the control group(all P＜0.05).Conclusion The established hepatocyte sphere co-culture model showed varying degrees of expression of phase Ⅰ/Ⅱ drug metabolism enzymes,transporters,and liver cell specific marker molecule albumin and can be used to evaluate the toxicity of multiflorum multiflorum,which provides further reference for the clinical application of multiflorum multiflorum.关键词
何首乌/肝毒性模型/共培养/3D细胞球Key words
Polygonum multiflorum/hepatotoxicity model/co-culture/3D cell spheres分类
中医学引用本文复制引用
苏华龙,姚香草,陈佳敏,岑柏宏,王萍,陈宗正,许重远..基于3D肝细胞球模型评价何首乌醇提物毒性研究[J].中国临床药理学杂志,2024,40(9):1272-1276,5.基金项目
广东省基础与应用基础研究基金企业联合基金资助项目(2022A1515220136) （2022A1515220136）
广东省研究生教育创新计划基金资助项目(2022ANLK_023) （2022ANLK_023）
青年基金-南方医科大学南方医院院长基金资助(2022B043) （2022B043）
南方医院临床研究专项基金资助项目(2019CR015) （2019CR015）
