Exploration of the toxic effects of ochratoxin A on HK-2 cells by optimizing serum level in cell culture medium
[Objectives]This paper aimed to investigate the effects of cell culture medium adding different levels of serum on ochratoxin A(OTA)toxicity to understand the toxicity of OTA better,and to provide an important reference for other studies in vitro.[Methods]The effect of OTA on the cell viability of human proximal tubule-derived epithelial cells(HK-2)was firstly determined,and OTA concentration in the cell supernatant was detected by ultra-high-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)in order to indirectly respond to the cellular uptake of OTA.The effect of adding different levels of serum in the culture medium on OTA toxicity was analyzed.[Results]The addition of 5％serum to the cell culture medium didn't affect the cell viability within 48 h and could be used as a normal cell culture condition;the addition of 2％serum didn't affect the cell viability at 36 h and could be used as a cell processing condition for the preparation of OTA working solution.Based on the optimized cell culture and treatment conditions,the effect results of OTA on the viability of HK-2 cells showed that the inhibition of OTA on HK-2 cells was time-dependent and dose-dependent.The IC50 of HK-2 cells treated by OTA for 12,24 and 36 h was 39.29,6.28 and 2.72 μmol·L-1,respectively.Meanwhile,OTA could also significantly increase the release level of lactate dehydrogenase in a time-dependent manner and damage the integrity of the cell membrane(P＜0.01).[Conclusions]The toxicity of OTA could be affected by the serum in the cell medium,and the higher the serum level,the greater the impact on OTA toxicity.When OTA working solutions was prepared using media with a 2％serum,the effect of serum level on OTA toxicity could be minimized without affecting cell viability.
ochratoxin Aserum levelHK-2 cellsultra-high-performance liquid chromatography-tandem mass spectrometry method(UHPLC-MS/MS)
万方数据
维普
