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Toxicity and Mechanism of Pyrazinamide in Human Hepatocyte L02
LIU Mengxing, LIU Yuan, SUN Hui, LIU Xing, YANG Min, PENG Jiangli, CHEN Jie
2021, 18(11):
1043-1047.
DOI: 10.19803/j.1672-8629.2021.11.10
Objectiv eTo evaluate the liver toxicity of pyrazinamide and to explore its mechanism of liver injury. Methods L02 cells were divided into the control group, PZA group (25, 125, 625, 3 125 µg/mL), and N-acetlg-L-cysteine (NAC) (10 mmol/L) + PZA (25, 125, 625, 3 125 µg/mL) group. The corresponding culture medium and different concentrations of PZA and NAC were given respectively. Flow cytometry was used to quantitatively detect the degree of apoptosis and intracellular mitochondrial membrane potential (MMP) of L02 cells. The Western-blot method was used to detect the protein expressions of Fas, Fas-related protein (FADD), cytochrome C (Cyt-c), and activated caspase-3 (Cleaved caspase-3). Results Compared with the control group, the proportion of apoptotic cells in the PZA administration group at each concentration was significantly increased, and the proportion of cells at the mid-late stage of apoptosis was significantly higher than that of cells at the early stage of apoptosis. MMP in the 25 and 125 µg/mL PZA administration groups was decreased by 32.23% and 24.79%, respectively, compared with 31.40% in the 3 125 µg/mL PZA administration group. The expression of Fas protein in the 3 125 µg/mL PZA administration group increased, so did the expression of Cleaved caspase-3 protein in the 125, 625 µg/mL PZA administration group. After PZA was combined with NAC, the proportion of apoptotic cells in each concentration group was not only lower than that of the PZA alone group, but decreased significantly in the 25, 125 and 3 125 μg/mL groups. Compared with the PZA alone group, MMP was increased by 21.95%, 33.48%, and 22.33% respectively in the 25, 625, and 3 125 µg/mL groups. Compared with the PZA alone group, Fas protein expression decreased by 47.50% in the 3 125 µg/mL group. The expression of Cleaved caspase-3 protein in the 125, 625, 3 125 µg/mL groups decreased by 66.13%, 73.91%, and 41.67%, respectively. Conclusion PZA may induce apoptosis of L02 cells through the death receptor pathway and mitochondrial pathway, resulting in liver injury. The combination with NAC can significantly alleviate liver injury induced by PZA.
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