OBJECTIVETo study the effect of excessive fluoride on calcium overload and apoptosis in cultured rat ameloblasts in vitro.

METHODSLogarithmic-phase ameloblasts (HAT-7) were treated with 0, 0.4, 0.8, 1.6, 3.2, and 6.4 mmol · L(-1) sodium fluoride (NaF) solution. Cell activities were detected by using a Cell Counting Kit 8 (CCK-8) assay after 48 h of treatment. The effect of fluoride on cell apoptosis was analyzed by using flow cytometry. Excessive fluoride-induced calcium concentration and calreticulin expression changes in ameloblasts were detected by using laser scanning confocal microscopy, Western blot analysis, and real-time quantitative polymerase chain reaction.

RESULTSNaF inhibited ameloblast activity at 1.6, 3.2, and 6.4 mmol · L(-1) (dose-dependent) after 48 h of induction. The Ca2+ fluorescence intensity of HAT-7 cells incubated with 1.6 and 3.2 mmol · L(-1) NaF was higher than that in the control group. The fluoride-induced early-stage apoptosis of ameloblasts after 48 h of induction and the early-stage apoptosis rate was positively correlated with fluoride concentration. Calreticulin mRNA expression in HAT-7 cells was higher than that in the control group after 48 h of incubation with 0.8, 1.2, and 1.6 mmol · L(-1) NaF.

CONCLUSIONExcessive fluoride-induced calcium overload in ameloblasts and further caused endoplasmic reticulum stress-mediated apoptosis.

Ameloblasts ; Animals ; Apoptosis ; Calcium ; Calcium Fluoride ; Fluorides ; Phosphates ; Rats ; Sodium Fluoride

Objective:To evaluate the effect of fluoride on the viability of rat ameloblast HAT-7 cells and calcium concentration in the cells.Methods:HAT-7 cells were exposed to NaF at 0,0.4,0.8,1.6,3.2 and 6.4 mmol/L for 24,48 and 72 h respectively. CCK-8 assay was performed to examine the cells proliferation;the apoptosis rate was determined by flow cytometry;Ca2 +concentration in the cells was detected by laser scanning confocal microscopy.Results:The cell proliferation was increased by NaF at 0.4 mmol/L and 0.8 mmol/L,whereas inhibited at 1.6 mmol/L and above.The effects were in a time-dependent manner.NaF increased apoptosis of the cells and increased Ca2 + concentration in the cells in a concentration-dependent manner.Conclusion:Fluoride at low doses promotes proliferation,at high doses inhibits proliferation of HAT-7 cells.NaF of 1.6 mmol/L or more induces apoptosis of HAT-7 cells and in-duce Ca2 + overloading in the cells.

Objective To study the effect of excessive fluoride on calcium overload and apoptosis in cultured rat ameloblasts in vitro. Methods Logarithmic-phase ameloblasts (HAT-7) were treated with 0, 0.4, 0.8, 1.6, 3.2, and 6.4 mmol·L-1 sodium fluoride (NaF) solution. Cell activities were detected by using a Cell Counting Kit 8 (CCK-8) assay after 48 h of treatment. The effect of fluoride on cell apoptosis was analyzed by using flow cytometry. Excessive fluoride-induced calcium concentration and calreticulin expression changes in ameloblasts were detected by using laser scanning confocal microscopy, Western blot analysis, and real-time quantitative polymerase chain reaction. Results NaF inhibited ameloblast activity at 1.6, 3.2, and 6.4 mmol·L?1 (dose-dependent) after 48 h of induction. The Ca2+ fluorescence intensity of HAT-7 cells incubated with 1.6 and 3.2 mmol·L?1 NaF was higher than that in the control group. The fluoride-induced early-stage apoptosis of ameloblasts after 48 h of induction and the early-stage apoptosis rate was positively correlated with fluoride concentration. Calreticulin mRNA expression in HAT-7 cells was higher than that in the control group after 48 h of incubation with 0.8, 1.2, and 1.6 mmol·L?1 NaF. Conclusion Excessive fluoride-induced calcium overload in ameloblasts and further caused endoplasmic reticulum stress-mediated apoptosis.

Sepsis is a prevalent critical illness observed in emergency intensive care unit (ICU), characterized by life-threatening organ dysfunction caused by infection-induced inflammatory immune disorders in the body. The suppression of immune function plays a crucial role in the development and progression of sepsis. Traditional Chinese medicine theory of "acute deficiency syndrome" in sepsis shares similarities with the concept of "immunosuppression". According to this theory, ginseng is frequently utilized in clinical treatment of sepsis due to its ability to invigorate vitality and strengthen the body, playing a crucial role in tonifying deficiency and improving the overall health of patients. This paper provides a detailed discussion of the pathophysiological mechanisms of sepsis immune dysfunction and its correlation with "acute deficiency syndrome" in traditional Chinese medicine. It summarizes the current state of modern pharmacological research on ginseng's impact on the body's immune function, discusses relevant research progress and shortcomings regarding ginseng's therapeutic effects on immunosuppression in sepsis, and proposes future research directions.