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. 2021 May 11;9:639944. doi: 10.3389/fcell.2021.639944

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Copyright © 2021 Shi, Wei, Xu, Liu, Zhang, Lv and Wang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Vitamin C causes oxidative stress in P. berghei–infected RBCs and parasites. (A) Vc induced mitochondrial superoxide production in a dose-dependent manner in P. berghei–infected RBCs, which could be reversed by the antioxidants N-acetyl cysteine (NAC) and glutathione (GSH). Left: Representative FACS plots of superoxide production in iRBCs. Middle: Quantitative superoxide production in iRBCs (n = 3/group). One-way ANOVA was performed to detect statistical differences. Right: Quantitative superoxide production in both iRBCs and RBCs (n = 3/group). Differences between Vc-treated iRBCs/RBCs and non-treated iRBCs/RBCs were analyzed by two-way ANOVA with Tukey multiple comparisons. (B) Vc induced superoxide production in a dose-dependent manner in P. berghei–infected RBCs measured by DHE, which could be reversed by NAC and GSH. Left: Representative FACS plots of superoxide production in iRBCs. Middle: Quantitative superoxide production in iRBCs (n = 3/group). Right: Quantitative superoxide production in both iRBCs and RBCs (n = 3/group). (C) Vc treatment decreased the GSH/GSSG ratio in P. berghei–infected RBCs (left panel) and isolated P. berghei parasites (right panel) (n = 5/group). (D) Vc decreased ATP production in P. berghei–infected RBCs (left panel) and isolated parasites (right panel) (n = 5/group). Based on calibration, the ATP concentrations of the tested samples derived from iRBCs and isolated parasites treated with 0 mM Vc are 5.40 ± 0.33 μM and 0.72 ± 0.003 μM. One-way ANOVA was performed to analyzed the results in (C, D). All data are presented as mean ± SD and are representative of at least three repetitions. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and n.s., not significant.