氟诱导大鼠口腔黏膜细胞和肝细胞的DNA损伤、凋亡及细胞周期变化。
DNA damage, apoptosis and cell cycle changes induced by fluoride in rat oral mucosal cells and hepatocytes.
作者信息
He Ling-Fei, Chen Jian-Gang
机构信息
Department of Dental Medicine, Zhongnan Hospital, Wuhan University, Wuhan 430071, Hubei Province, China.
出版信息
World J Gastroenterol. 2006 Feb 21;12(7):1144-8. doi: 10.3748/wjg.v12.i7.1144.
AIM
To study the effect of fluoride on oxidative stress, DNA damage and apoptosis as well as cell cycle of rat oral mucosal cells and hepatocytes.
METHODS
Ten male SD rats weighing 80-120 g were randomly divided into control group and fluoride group, 5 animals each group. The animals in fluoride group had free access to deionized water containing 150 mg/L sodium fluoride (NaF). The animals in control group were given distilled water. Four weeks later, the animals were killed. Reactive oxygen species (ROS) in oral mucosa and liver were measured by Fenton reaction, lipid peroxidation product, malondialdehyde (MDA), was detected by thiobarbituric acid (TBA) reaction, reduced glutathione (GSH) was assayed by dithionitrobenzoic acid (DTNB) reaction. DNA damage in oral mucosal cells and hepatocytes was determined by single cell gel (SCG) electrophoresis or comet assay. Apoptosis and cell cycle in oral mucosal cells and hepatocytes were detected by flow cytometry.
RESULTS
The contents of ROS and MDA in oral mucosa and liver tissue of fluoride group were significantly higher than those of control group (P < 0.01), but the level of GSH was markedly decreased (P < 0.01). The contents of ROS, MDA and GSH were (134.73 +/- 12.63) U/mg protein, (1.48 +/- 0.13) mmol/mg protein and (76.38 +/- 6.71) mmol/mg protein in oral mucosa respectively, and (143.45 +/- 11.76) U/mg protein, (1.44 +/- 0.12) mmol/mg protein and (78.83 +/- 7.72) mmol/mg protein in liver tissue respectively. The DNA damage rate in fluoride group was 50.20% in oral mucosal cells and 44.80% in hepatocytes, higher than those in the control group (P < 0.01). The apoptosis rate in oral mucosal cells was (13.63 +/- 1.81) % in fluoride group, and (12.76 +/- 1.67)% in hepatocytes, higher than those in control group. Excess fluoride could differently lower the number of oral mucosal cells and hepatocytes at G0/G1 and S G2/M phases (P < 0.05).
CONCLUSION
Excess fluoride can induce oxidative stress and DNA damage and lead to apoptosis and cell cycle change in rat oral mucosal cells and hepatocytes.
目的
研究氟对大鼠口腔黏膜细胞和肝细胞氧化应激、DNA损伤、凋亡以及细胞周期的影响。
方法
将10只体重80 - 120 g的雄性SD大鼠随机分为对照组和氟组，每组5只。氟组动物自由饮用含150 mg/L氟化钠（NaF）的去离子水，对照组动物给予蒸馏水。4周后，处死动物。采用Fenton反应测定口腔黏膜和肝脏中的活性氧（ROS），用硫代巴比妥酸（TBA）反应检测脂质过氧化产物丙二醛（MDA），用二硫代硝基苯甲酸（DTNB）反应测定还原型谷胱甘肽（GSH）。通过单细胞凝胶（SCG）电泳或彗星试验检测口腔黏膜细胞和肝细胞中的DNA损伤。采用流式细胞术检测口腔黏膜细胞和肝细胞的凋亡及细胞周期。
结果
氟组口腔黏膜和肝脏组织中ROS和MDA含量显著高于对照组（P < 0.01），但GSH水平明显降低（P < 0.01）。口腔黏膜中ROS、MDA和GSH含量分别为（134.73 ± 12.63）U/mg蛋白、（1.48 ± 0.13）mmol/mg蛋白和（76.38 ± 6.71）mmol/mg蛋白，肝脏组织中分别为（143.45 ± 11.76）U/mg蛋白、（1.44 ± 0.12）mmol/mg蛋白和（78.83 ± 7.72）mmol/mg蛋白。氟组口腔黏膜细胞DNA损伤率为50.20%，肝细胞为44.80%，高于对照组（P < 0.01）。氟组口腔黏膜细胞凋亡率为（13.63 ± 1.81）%，肝细胞为（12.76 ± 1.67）%，高于对照组。过量氟可不同程度降低口腔黏膜细胞和肝细胞在G0/G1期及S + G2/M期的细胞数量（P < 0.05）。
结论
过量氟可诱导大鼠口腔黏膜细胞和肝细胞氧化应激及DNA损伤，导致细胞凋亡和细胞周期改变。
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