4种有机溶剂对Vero细胞活力和弓形虫增殖的影响

doi:10.16431/j.cnki.1671-7236.2019.08.009

摘要/Abstract

摘要：

本研究旨在筛选低细胞毒性和低弓形虫毒性的有机溶剂。Vero细胞在96孔板培养12 h后，将培养基更换为200μL分别含0、0.1%、0.3%、0.5%、1%、3%、5%、10%（V/V）的二甲基亚砜（DMSO）、二甲基甲酰胺、乙醇和甲醇的完全培养基，同时设立空白对照孔，继续培养12和24 h后，采用噻唑蓝（MTT）法评价4种有机溶剂对Vero细胞增殖活力的影响，筛选出对宿主细胞毒性最小的有机溶剂并确定其安全浓度；弓形虫接种Vero细胞12 h后，弃上清后分别加入含有0、0.1%、0.3%、0.5%、1%、3%（V/V）的DMSO、二甲基甲酰胺、乙醇和甲醇的完全培养基120μL，接种36 h后判定各组溶剂对弓形虫增殖的影响，计数感染细胞数并计算相对感染率，筛选出对弓形虫增殖影响最小的有机溶剂及其安全浓度。结果显示，对于Vero细胞，DMSO、二甲基甲酰胺、甲醇和乙醇分别在其体积分数低于1%、0.1%、3%和1%时无明显毒性；对于弓形虫，DMSO、二甲基甲酰胺、甲醇和乙醇对弓形虫增殖的安全添加浓度分别为1%、0.1%、1%和0.1%。本研究表明，DMSO、甲醇对Vero细胞、弓形虫的毒性较小，可作为水溶性较差药物进行抗弓形虫筛选时优先选择的有机溶剂。

关键词: 有机溶剂; Vero细胞; 弓形虫; 活力; 增殖

Abstract:

The aim of this study was to screen the organic solvents with lower cytotoxicity and Toxoplasma gondi toxicity.Vero cells were cultured on 96-well plates, the cells were cultured for 12 hours and then the medium was replaced by 200 μL complete DMEM medium containing 0, 0.1%, 0.3%, 0.5%, 1%, 3%, 5% and 10% (V/V) DMSO, dimethyformamide, ethanol and methanol, respectively, blank control holes were set up at the same time.The toxicity of DMSO, dimethylformamide, methanol and ethanol to Vero cells was analyzed by MTT method.The organic solvents with the least cytotoxicity to host cells were screened and the safe concentration was determined.Toxoplasma gondi was inoculated to Vero cells and cultured for 12 hours.The supernatant was discarded and then 120 μL complete DMEM containing 0, 0.1%, 0.3%, 0.5%, 1% and 3% (V/V) DMSO, dimethyformamide, ethanol and methanol was added, respectively.The impacts of solvents on the proliferation of Toxoplasma gondi were determined 36 hours post inoculation.The number of infected cells was counted and the relative infection rate was calculated.The organic solvents with the least impact on the proliferation of Toxoplasma gondi and the safe concentration were screened out.The results showed that there was no obvious cytotoxicity on Vero cells when the volume fraction of DMSO, dimethylformamide, methanol and ethanol were less than 1%, 0.1%, 3% and 1%, respectively.For Toxoplasma gondi, the safety concentrations of DMSO, dimethylformamide, methanol and ethanol were 1%, 0.1%, 1% and 0.1%, respectively.This results indicated DMSO and methanol were less toxic to Vero cells and Toxoplasma gondii, and could be used as organic solvents for the screening of drugs with poor water solubility against Toxoplasma gondii.

Key words: organic solvents; Vero cells; Toxoplasma gondii; vitality; proliferation

中图分类号:

S851.34⁺7

薛俊欣, 熊炜, 张强, 黄忠荣, 李树清, 李春阳, 赵新宇, 李健, 王权. 4种有机溶剂对Vero细胞活力和弓形虫增殖的影响[J]. 《中国畜牧兽医》, 2019, 46(8): 2265-2272.

XUE Junxin, XIONG Wei, ZHANG Qiang, HUANG Zhongrong, LI Shuqing, LI Chunyang, ZHAO Xinyu, LI Jian, WANG Quan. Influence of Four Organic Solvents on Viability of Vero Cells and Proliferation of Toxoplasma gondii[J]. , 2019, 46(8): 2265-2272.

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