Corrigendum: Mechanisms by which statins protect endothelial cells from radiation-induced injury in the carotid artery

Abstract

[This corrects the article DOI: 10.3389/fcvm.2023.1133315.].

Keywords: carotid stenosis; endothelium; mitochondria; prevention; radiation therapy; statin.

Figure 1

Pravastatin and atorvastatin preserve endothelial function in vivo following head-and-neck IR. (A–C) Effects of statins on endothelium-dependent relaxation of the carotid artery in response to acetylcholine (ACh). C57BL/6J mice were treated with (A) vehicle, (B) pravastatin (Prava), or (C) atorvastatin (Ator) after head-and-neck irradiation (12Gy) or sham treatment, and relaxation was tested at 24 and 240 h after irradiation. (D–F) Effects of statins on endothelium-independent relaxation of the carotid artery in response to sodium nitroprusside (SNP), in mice treated as in A, B, and C, respectively. (G–I) Effects of statins on endothelium-dependent relaxation of mesenteric resistance arteries (MRAs), in mice treated as in A, B, and C, respectively. n = 5 mice per group. p values were determined using repeated measures 2-way ANOVA followed by Tukey's post-hoc test.

Figure 3

Pravastatin protects against IR-induced mitochondrial damage or hyperpolarization in vitro. All panels compare cells subjected to irradiation (4Gy) after pretreatment with pravastatin (Prava, 10 μM) starting at 18 h before irradiation. (A,B) Representative images and signal integrated density of mitoSOX fluorescence normalized to mitoTracker fluorescence in HCAECs at (A) 24 and (B) 240 h after IR. (C,D) _mtDNA lesions in DNA extracted from HUVECs at (C) 24 h and (D) 240 h after IR. (E,F) Representative images and integrated density of mitochondrial membrane potential in HCAECs, as determined by TMRM fluorescence, at (E) 24 h and (F) 240 h after irradiation. Analysis per cell, n = 4 independent experiments. p values were determined by Kruskal–Wallis test.

Figure 4

Atorvastatin does not protect against IR-induced mitochondrial damage or hyperpolarization in vitro. All panels compare HCAECs subjected to irradiation (4 Gy) after pretreatment with atorvastatin (Ator, 5 μM) or vehicle. Parameters assessed are: (A,B) Representative images and signal integrated density of MitoSOX fluorescence normalized to MitoTracker fluorescence at (A) 24 and (B) 240 h after irradiation, in cells treated with atorvastatin (5 μM) or vehicle starting 18hr before irradiation. (C,D) Damage to _mtDNA as assessed by PCR assay. _mtDNA lesions at (C) 24 and (D) 240 h after irradiation, in HUVECs treated with atorvastatin or vehicle starting 18 h before IR. (E,F) Representative images and integrated density of mitochondrial membrane potential, as determined by TMRM fluorescence, at (E) 24 and (F) 240 h after irradiation. Analysis per cell, n = 4 independent experiments, p values by Kruskal–Wallis test.

Figure 7

Pravastatin, but not atorvastatin, prevents irradiation-induced reduction of _mtDNA transcription and ETC activity. All panels compare HCAECs subjected to irradiation (4Gy) after pretreatment with atorvastatin (Ator, 5 μM, overnight), pravastatin (Prava, 10 μM, overnight) or vehicle. (A–D) Effects of pretreatment with pravastatin (Prava, 10 μM, overnight) on transcriptional activity. (A,B) Quantitative (q)RT-PCR for cytochrome c oxidase I (MT-COI) at (A) 24 and (B) 240 h after irradiation. (C–D) qRT-PCR for NADH-ubiquinone oxidoreductase chain 1 (MT-ND1) at (C) 24 and (D) 240 h after irradiation. (E–H) Effects of pretreatment with atorvastatin (Ator, 5 μM, overnight) on transcriptional activity. (E,F) qRT-PCR for MT-COI at (E) 24 and (F) 240 h after irradiation. (G,H) qRT-PCR for MT-ND1 at (G) 24 and (H) 240 h after irradiation. (I,J) Activity of ETC complex 1, as assessed by fluorometric assay at (I) 24 and (J) 240 h after irradiation. p values by Kruskal–Wallis test.

Supplementary Figure 3

Neither pravastatin nor atorvastatin affects IR-induced transcription of nuclear DNA. (A–D) Effects of pretreatment with pravastatin (Prava, 10 μM, 1 h) on nucDNA damage in HCAECs after irradiation (IR, 4 Gy). (A,B) Quantitative (q)RT-PCR for NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 [(B), NDUF1], with cDNA normalized to 100 ng at 24 and 240 h after IR. (C,D) qRT-PCR for cytochrome c oxidase 11 [(D), COX11], with cDNA normalized to 100 ng at 24 and 240 h after IR. (E–H) Effects of pretreatment with atorvastatin (5 μM, overnight) on nucDNA damage in HCAECs subjected to IR. (E,F) qRT-PCR for NDUF1 (B), with cDNA normalized to 100 ng at 24 and 240 h after IR. (G,H) qRT-PCR for COX11 (D), with cDNA normalized to 100 ng at 24 and 240 h after IR. Statistical analysis by Kruskal–Wallis test. Ns indicates not significant.