Abstract
ObjectiveThis study aims to analyze the expressions of autophagy-related factors light chain 3 alpha (LC3A) and Beclin 1 and apoptosis-related factors B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (BAX) in primary osteoblasts treated with sodium fluoride (NaF).MethodsOsteoblasts were extracted from Sprague-Dawley rats and treated with 0, 2.5, 5, and 10 mg/L NaF solutions, followed by 10 mmol/L 3-methyladenine (3-MA) for 24 h. The apoptotic rate was determined by flow cytometry, and the expressions of the autophagy- and apoptosis-related factors were measured by western blotting and real-time quantitative polymerase chain reaction.ResultsThe mRNA expressions of LC3A, Beclin 1, and BAX in the NaF-treated osteoblast group were higher than those in the control group, while the protein expressions of these factors in the NaF-treated group were significantly higher than those in the control group. However, the Bcl-2 protein expression in the NaF-treated osteoblasts was significantly decreased compared to that in the control cells. After the 3-MA treatment, the protein expressions of LC3A, Beclin 1, and Bcl-2 were significantly decreased compared with those of the NaF-treated group, whereas the expression of BAX increased. Moreover, the apoptosis rate was increased after the addition of the 3-MA inhibitor.ConclusionNaF stimulation promoted autophagy and apoptosis of the osteoblasts, suggesting the involvement of fluoride damage in these processes.
Highlights
Endemic fluorosis is a global disease (Wei et al, 2019)
Light chain 3 alpha (LC3A) and Beclin 1 have been used as markers to detect autophagy activity (Ding et al, 2016)
When the primary cells were transferred to the fourth generation, the osteoblasts adhered to the wall with a good spindle shape, round nucleus, and good refractive properties
Summary
Endemic fluorosis is a global disease (Wei et al, 2019). While physiological doses of fluoride are beneficial to the human body and can promote bone growth and development, excessive fluoride intake leads to its accumulation in a variety of organs, eventually causing fluorosis (Wang et al, 2019a). Autophagy is a highly valued protective mechanism in the process of evolution (Antonioli et al, 2017; Ommati et al, 2019, 2020a,b) While it can remove damaged proteins and organelles to maintain the stability of the intracellular environment, in excess it can cause damage to organelles, an unstable internal environment, and eventually death (Cicchini et al, 2015). This may be one of the reasons for the pathological changes of osteomalacia, osteosclerosis, and osteoporosis in fluorosis. Apoptosis is regulated by many related genes, such as B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (BAX) (Anesan et al, 2018)
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