官志忠,王亚南,Gustav Dallner
GUAN Zhi-zhong 1, WANG Ya-nan 2,3, Gustav Dallner 3 (1. Molecular Biological Key Laboratory of Guiyang Medical College, Guiyang 550004, China; 2. Research Administration Department, Guiyang Medical College, Guiyang 550004, China; 3. Department of Biophysics and Biochemistry, Stockholm University, 14186 Stockholm, Sweden)

• 1:贵阳医学院分子生物学重点实验室
• 2:贵阳医学院科研处
• 3:Stockholm大学生物物理学与生物化学系 贵州贵阳550004
• 4:贵州贵阳550004

摘要(Abstract)：

目的:深入了解氟中毒对神经细胞中生物膜性脂质结构的影响及其改变机制。方法:体外培养SH SY5Y人脑神经母细胞瘤细胞,在培养液中加入不同浓度的氟化物或加入抗氧化剂,培养48h后用测定细胞 MTT的方法来了解细胞的损伤程度,用高效液相色谱法分离和测定培养液中脂质过氧化物水平,用过柱和比色 法测定细胞生物膜磷脂含量,用高效液相色谱法测定细胞生物膜辅酶Q和胆固醇含量。结果:经氟处理的神经 细胞中MTT水平降低和脂质过氧化水平升高,呈现剂量依赖关系;高浓度氟处理的神经细胞中磷脂和辅酶Q含 量减少,但胆固醇含量不变;加入维生素E后可减弱高剂量氟对生物膜性脂质的破坏作用。结论:氟中毒可引 起SH SY5Y神经母细胞瘤细胞生物膜性脂质结构发生异常改变,其机制可能与高剂量氟引起的自由基增多而 对细胞生物膜造成的损害有关。
Objective: To investigate the influence of fluoride on cellular membrane lipids in SH-SY5Y cells as well as to reveal mechanism of lipid modification. Methods: SH-SY5Y cells, a human neuroblastoma cell line, were incubated with different concentrations of fluoride for 48 hr and somle of them were treated with vitamin E precedently. The functional situation of cells was measured by MTT method; lipid peroxidation was detected by High-Performance Liquid Chromatography (HPLC); phospholipid was separated by a Silica SepPak cartridge and neutral lipids by HPLC. Results: In the groups with high-doses of fluoride as compared to control, the decreased MTT and increased lipid peroxidation were obviously observed in SH-SY5Y cells with a manner of opposite concentration-dependence. Decreases of phospholipid and ubiquinone, but not cholesterol, were found in the cells treated with high-doses of fluoride. Prior treatment with vitamin E can prevent the decreases of these lipids in the cells exposed to high dose of fluoride. Conclusion: High-doses of fluoride can induce modification of membrane lipids in SH-SY5Y cells, which might be involved in a mechanism of free radical attack resulted from fluorosis.

关键词(KeyWords)： 氟化物中毒;神经母细胞瘤;磷脂类;泛醌;胆固醇;脂质过氧化作用
fluoride poisoning; neuroblastoma; phospholipids; ubiquinone; cholesterol; lipid peroxidation

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(30060026);; 贵州省省长基金资助项目

作者(Author): 官志忠,王亚南,Gustav Dallner
GUAN Zhi-zhong 1, WANG Ya-nan 2,3, Gustav Dallner 3 (1. Molecular Biological Key Laboratory of Guiyang Medical College, Guiyang 550004, China; 2. Research Administration Department, Guiyang Medical College, Guiyang 550004, China; 3. Department of Biophysics and Biochemistry, Stockholm University, 14186 Stockholm, Sweden)

Email:

DOI: 10.19367/j.cnki.1000-2707.2005.01.001

参考文献(References)：

• [1]MulienixPJ,DenbestenPK,SchuniorA,etal.Neurotox icityofsodiumfluorideinrats[J].Neurotoxicology&Ter atology,1995,17:169-177.
• [2]GuanZZ,WangYN,XiaoKQ,etal.Influenceofchronic fluorosisonmembranelipidsinratbrain[J].Neurotoxi cology&Teratology,1998,20:537-542.
• [3]LiuY,PetersonDA,KimuraH,etal.Mechanismofcel lular3(4,5 dimethylthiazol2 yl)2,5 diphenyltetrazoli umbromide(MTT)reduction[J].JournalofNeurochem istry,1997,69:581-593.
• [4]DraperHH,SquiresEJ,MahmoodiH,etal.Acompara tiveevaluationofthiobarbituricacidmethodsforthedeter minationofmalondialdehydeinbiologicalmaterials[J].FreeRadicalBiology&Medicine,1993,15:353-363.
• [5]GuanZZ,GrunlerJ,PiaoS,etal.Separationandquanti tationofphospholipidsandtheiretheranaloguesbyhigh performanceliquidchromatography[J].AnalyticalBio chemistry,2001,297:137-143.
• [6]GuanZZ,XiaoKQ,ZengXY,etal.Changedcellular membranelipidcompositionandlipidperoxidationofkid neyinratswithchronicfluorosis[J].ArchiveToxicology,2000,74:602-608.
• [7]WangYN,XiaoKQ,LiuJL,etal.Effectoflongterm fluorideexposureonlipidcompositioninratliver[J].Toxicology,2000,146:161-169.

文章评论(Comment)：