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目的探讨硒对氟诱导的原代大鼠肝细胞氧化应激和DNA损伤的拮抗作用。方法采用改良原位2步非循环灌流法及多次过滤低速离心法分离原代大鼠肝细胞,实验设空白对照组、氟染毒组(氟化钠250、500、1 000、2 000μmol/L,染毒24 h)和硒干预组(亚硒酸钠10或100 nmol/L,干预4 h);测定细胞内活性氧(ROS)、丙二醛(MDA)、谷胱甘肽(GSH)含量及超氧化物歧化酶(SOD)活力,采用单细胞凝胶电泳技术和微核实验检测DNA损伤情况。结果与空白对照组比较,2 000μmol/L氟染毒组ROS(16.75±8.32)和MDA[(11.54±3.83)nmol/mg·prot]含量明显增加,SOD[(19.53±5.28)NU/mg·prot]和GSH[(21.73±15.32)μg/mg·prot]明显下降,彗星率[(46.25±10.60)%]、彗星尾长[(31.74±9.25)μm]、尾矩[(15.57±7.92)]和微核率[(42.80±24.61)‰]明显增加(均P<0.01);与等剂量氟染毒组比较,10 nmol/L硒干预组ROS(12.36±5.15)和MDA[(7.33±4.01)nmol/mg·prot]含量明显下降,SOD[(23.72±7.15)NU/mg·prot]与GSH[(26.36±14.24)μg/mg·prot]明显增加,彗星率[(39.27±15.0)%]、彗星尾长[(24.06±8.77)μm]、尾矩[(9.64±4.80)]和微核率[(31.28±12.65)‰]分别降低(P<0.05)。结论适量的硒可拮抗氟对原代大鼠肝细胞的氧化应激和DNA损伤作用。
Objective To explore the antagonistic effect of selenium on fluoride-induced oxidative stress and DNA damage in primary rat hepatocytes. Methods The primary rat hepatocytes were isolated by modified two-step non-perfusion in situ flow cytometry and multiple filtration low-speed centrifugation. The experimental groups were divided into blank control group, fluorosis group (250, 500, 1000 and 2000μmol / L and exposed to selenium for 24 h) and selenium intervention group (10 or 100 nmol / L sodium selenite for 4 h). The levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione ) Content and superoxide dismutase (SOD) activity were detected by single cell gel electrophoresis and micronucleus test DNA damage. Results Compared with the blank control group, ROS (16.75 ± 8.32) and MDA [(11.54 ± 3.83) nmol / mg · prot] significantly increased in the 2 000 μmol / (21.73 ± 15.32) μg / mg · prot], the comet rate [(46.25 ± 10.60)%], the tail length of tail [(31.74 ± 9.25) μm] and the tail moment [(15.57 ± 7.92) (P <0.01). Compared with the same dose of fluoride exposure group, ROS (12.36 ± 5.15) and MDA [(7.33 ± 4.01) nmol / mg · prot] significantly decreased with the increase of SOD [(23.72 ± 7.15) NU / mg · prot] and GSH [(26.36 ± 14.24) μg / mg · prot] %], Tail length (24.06 ± 8.77 μm), tail moment (9.64 ± 4.80) and micronucleus rate (31.28 ± 12.65)%, respectively. Conclusions Selenium can antagonize the oxidative stress and DNA damage of primary rat hepatocytes induced by fluoride.