In Vitro Evaluation of the Apoptotic, Autophagic, and Necrotic Molecular Pathways of Fluoride
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In Vitro Evaluation of the Apoptotic, Autophagic, and Necrotic Molecular Pathways of Fluoride F. Urut 1,2,3,4 & S. Dede 1,2,3,4
&
V. Yuksek 1,2,3,4 & S. Cetin 1,2,3,4 & A. Usta 1,2,3,4 & M. Taspinar 1,2,3,4
Received: 20 October 2020 / Accepted: 9 November 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Prolonged exposure to high doses of fluoride causes chronic poisoning called fluorosis, which affects many tissues and causes serious health problems. This study was planned to investigate the apoptotic, autophagic, and necrotic molecular pathways of fluoride. Sodium fluoride (NaF) was administered to normal rat kidney epithelial (NRK-52E) cells. The NaF IC50 value was determined using the MTT assay. The expression of the genes in the autophagic, apoptotic, and necrotic pathways was determined by real-time PCR. It was determined that there were significant changes in NaF-induced molecular pathways depending on the time. There were no increases in apoptotic and necrotic pathway markers except for Atg3, an autophagy gene, at the 3rd and the 12th hours. However, there was an induction in all cell death signaling pathways at 24 h. The molecular mechanisms demonstrated NaF-induced cellular death in the NRK-52E cell line. It was concluded that these molecular mechanisms were activated with NaF, and different mechanisms accelerated the cellular death at the 24th hour. Keywords NaF . NRK-52E cell line . In vitro . Apoptosis . Autophagy . Necrosis
Introduction Fluoride (F) is an element with high electronegativity that can be naturally found in soil, water, and various nutrients and many products used in daily life (toothpaste, insecticides, some chemical compounds). Fluoride toxicity, known as fluorosis, occurs in humans and animals due to long-term exposure to high concentrations of fluoride. In fluorosis, several tissues, especially the teeth and the bones, are damaged. Fluoride is mainly excreted through the kidneys. Since the kidney is the primary organ responsible for fluoride excretion, effects on the kidneys are expected. During the filtration of fluoride, the kidneys are exposed to large amounts of fluoride
* S. Dede [email protected] 1
Biochemistry Department, Faculty of Veterinary Medicine, Van Yuzuncu Yil University, 65090 Van, Turkey
2
Özalp Regional High School, Van Yuzuncu Yil University, 65090 Van, Turkey
3
Chemistry Department, Science Faculty, Van Yuzuncu Yil University, 65090 Van, Turkey
4
Medical Biology Department, Medical Faculty, Aksaray University, Aksaray, Turkey
and participate in fluoride homeostasis with calcified tissues. Fluoride can lead to necrosis through various cellular mechanisms; however, its role has not been fully clarified [1–4]. Several studies have been carried out to clarify fluorideinduced toxicity mechanisms. Oxidative stress–induced DNA damage and activation of apoptotic pathways are important mechanisms [3–6]. Apoptosis is necessary for the normal functions and survival of multicellular organisms. Each cell lives for a certain perio
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