Toxicity comparison of psoralen in 2D /3D hepatocyte models and its effect on mitochondrial fusion and fission / 中国药理学通报

ABSTRACT

Aim To evaluate and compare the toxicity of psoralen on two-dimensional(2D)and three-dimensional(3D)cultured human hepatocyte HepG2 models. Methods The 3D cell model of HepG2 cells was constructed by low adsorption U-shaped bottom porous plate method. After treated with psoralen for 24 hours, the cell viabilities of 2D and 3D HepG2 cells were detected by CCK-8 assay, LDH leakage was detected by kit, and the mitochondrial membrane potential was detected by TMRM staining. The effect of psoralen on the mRNA of mitochondrial fusion-fission proteins DRP1, Mfn-2 and OPA1 was detected by Q-PCR. Results 3D model maintained a high level of albumin and urea secretion for a long time. And the expression levels of CYP1A2, CYP2E1, CYP3A4 and UGT1A1 in 3D model were higher than those in 2D model. In 3D model lower concentrations of psoralen showed a significant decrease in cell viability, a significant increase in LDH leakage, and a decrease in mitochondrial membrane potential. Q-PCR results showed that psoralen induced a marked increase in the expression of mitochondrial fission protein DRP1, while a significant decrease in mitochondrial fusion protein OPA1. Conclusions A 3D HepG2 cell model is successfully constructed and applied to the evaluation of psoralen hepatotoxicity; the 3D cell culture model is more sensitive to psoralen toxicity, and mitochondria may play a key role in psoralen-induced cell damage.