Mitochondrial Toxicity of Azithromycin Results in Aerobic Glycolysis and DNA Damage of Human Mammary Epithelia and Fibroblasts
Mitochondria evolved from free-living bacteria via endocytosis within eukaryotic host cells millions of year ago. We hypothesized that antibiotics cause mammalian mitochondrial damage while causing bacterial lethality. Mitochondrial toxicity of azithromycin in human mammary epithelia MCF-12A and fib...
Main Authors: | , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2019-08-01
|
Series: | Antibiotics |
Subjects: | |
Online Access: | https://www.mdpi.com/2079-6382/8/3/110 |
id |
doaj-ef38a8e3ff9b47ffad24b2efe83b0aaf |
---|---|
record_format |
Article |
spelling |
doaj-ef38a8e3ff9b47ffad24b2efe83b0aaf2020-11-24T21:34:29ZengMDPI AGAntibiotics2079-63822019-08-018311010.3390/antibiotics8030110antibiotics8030110Mitochondrial Toxicity of Azithromycin Results in Aerobic Glycolysis and DNA Damage of Human Mammary Epithelia and FibroblastsXianpeng Jiang0Catherine Baucom1Robert L. Elliott2Breast Cancer Research, Sallie A. Burdine Breast Foundation, Baton Rouge, LA 70806, USAElliott Mastology Center, Baton Rouge, LA 70806, USAElliott Mastology Center, Baton Rouge, LA 70806, USAMitochondria evolved from free-living bacteria via endocytosis within eukaryotic host cells millions of year ago. We hypothesized that antibiotics cause mammalian mitochondrial damage while causing bacterial lethality. Mitochondrial toxicity of azithromycin in human mammary epithelia MCF-12A and fibroblasts were tested by fluorescent and transmission electron microscopy. Gene expression and DNA damage were tested by real-time polymerase chain reaction (qPCR) and ELISA. We found azithromycin suppressed the mitochondrial membrane potential gradient of MCF-12A cells and fibroblasts. Ultrastructure exams showed that the antibiotic caused vacuolated and swollen mitochondria with disrupted cristae in MCF-12A cells and fibroblasts compared to the morphology of mitochondria in the cells without antibiotic treatment. Fluorescent microscopy also showed azithromycin-induced mitochondrial reactive oxygen species (ROS), superoxide, after 3 h of culture. The DNA oxidative damage product, 8-hydroxy-2’-deoxyguanosine (8-OHdG, significantly increased in the media after MCF-12A cells and fibroblasts were cultured in the media containing azithromycin for 24 h. Azithromycin upregulated gene expression of hypoxia inducible factor 1 alpha (<i>HIF1a</i>), glycolytic enzymes including hexokinase 2 (<i>HK2</i>), phosphofructokinase 1 (<i>PFKM</i>), pyruvate kinase muscle isozyme M2 (<i>PKM2</i>), and glucose transporters in MCF-12A cells and fibroblasts. Lactate production also increased in the culture media. After treatment with azithromycin, healthy MCF-12A and fibroblast cells increased aerobic glycolysis—the “Warburg Effect”—to generate energy. In summary, azithromycin caused mitochondrial toxicity, ROS overproduction, DNA oxidative damage, upregulation of the <i>HIF1a</i> gene, and aerobic glycolysis in healthy mammalian cells. Over-usage of antibiotics could contribute to tumorigenesis and neurodegeneration and aggravate existing mitochondria-associated diseases.https://www.mdpi.com/2079-6382/8/3/110azithromycinmitochondrial toxicityaerobic glycolysisDNA damage |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Xianpeng Jiang Catherine Baucom Robert L. Elliott |
spellingShingle |
Xianpeng Jiang Catherine Baucom Robert L. Elliott Mitochondrial Toxicity of Azithromycin Results in Aerobic Glycolysis and DNA Damage of Human Mammary Epithelia and Fibroblasts Antibiotics azithromycin mitochondrial toxicity aerobic glycolysis DNA damage |
author_facet |
Xianpeng Jiang Catherine Baucom Robert L. Elliott |
author_sort |
Xianpeng Jiang |
title |
Mitochondrial Toxicity of Azithromycin Results in Aerobic Glycolysis and DNA Damage of Human Mammary Epithelia and Fibroblasts |
title_short |
Mitochondrial Toxicity of Azithromycin Results in Aerobic Glycolysis and DNA Damage of Human Mammary Epithelia and Fibroblasts |
title_full |
Mitochondrial Toxicity of Azithromycin Results in Aerobic Glycolysis and DNA Damage of Human Mammary Epithelia and Fibroblasts |
title_fullStr |
Mitochondrial Toxicity of Azithromycin Results in Aerobic Glycolysis and DNA Damage of Human Mammary Epithelia and Fibroblasts |
title_full_unstemmed |
Mitochondrial Toxicity of Azithromycin Results in Aerobic Glycolysis and DNA Damage of Human Mammary Epithelia and Fibroblasts |
title_sort |
mitochondrial toxicity of azithromycin results in aerobic glycolysis and dna damage of human mammary epithelia and fibroblasts |
publisher |
MDPI AG |
series |
Antibiotics |
issn |
2079-6382 |
publishDate |
2019-08-01 |
description |
Mitochondria evolved from free-living bacteria via endocytosis within eukaryotic host cells millions of year ago. We hypothesized that antibiotics cause mammalian mitochondrial damage while causing bacterial lethality. Mitochondrial toxicity of azithromycin in human mammary epithelia MCF-12A and fibroblasts were tested by fluorescent and transmission electron microscopy. Gene expression and DNA damage were tested by real-time polymerase chain reaction (qPCR) and ELISA. We found azithromycin suppressed the mitochondrial membrane potential gradient of MCF-12A cells and fibroblasts. Ultrastructure exams showed that the antibiotic caused vacuolated and swollen mitochondria with disrupted cristae in MCF-12A cells and fibroblasts compared to the morphology of mitochondria in the cells without antibiotic treatment. Fluorescent microscopy also showed azithromycin-induced mitochondrial reactive oxygen species (ROS), superoxide, after 3 h of culture. The DNA oxidative damage product, 8-hydroxy-2’-deoxyguanosine (8-OHdG, significantly increased in the media after MCF-12A cells and fibroblasts were cultured in the media containing azithromycin for 24 h. Azithromycin upregulated gene expression of hypoxia inducible factor 1 alpha (<i>HIF1a</i>), glycolytic enzymes including hexokinase 2 (<i>HK2</i>), phosphofructokinase 1 (<i>PFKM</i>), pyruvate kinase muscle isozyme M2 (<i>PKM2</i>), and glucose transporters in MCF-12A cells and fibroblasts. Lactate production also increased in the culture media. After treatment with azithromycin, healthy MCF-12A and fibroblast cells increased aerobic glycolysis—the “Warburg Effect”—to generate energy. In summary, azithromycin caused mitochondrial toxicity, ROS overproduction, DNA oxidative damage, upregulation of the <i>HIF1a</i> gene, and aerobic glycolysis in healthy mammalian cells. Over-usage of antibiotics could contribute to tumorigenesis and neurodegeneration and aggravate existing mitochondria-associated diseases. |
topic |
azithromycin mitochondrial toxicity aerobic glycolysis DNA damage |
url |
https://www.mdpi.com/2079-6382/8/3/110 |
work_keys_str_mv |
AT xianpengjiang mitochondrialtoxicityofazithromycinresultsinaerobicglycolysisanddnadamageofhumanmammaryepitheliaandfibroblasts AT catherinebaucom mitochondrialtoxicityofazithromycinresultsinaerobicglycolysisanddnadamageofhumanmammaryepitheliaandfibroblasts AT robertlelliott mitochondrialtoxicityofazithromycinresultsinaerobicglycolysisanddnadamageofhumanmammaryepitheliaandfibroblasts |
_version_ |
1725949183342411776 |