【摘要】：實(shí)驗(yàn)?zāi)康模?研究工頻磁場暴露對FL細(xì)胞內(nèi)神經(jīng)酰胺代謝的影響。 實(shí)驗(yàn)方法： 人羊膜(FL)細(xì)胞經(jīng)50Hz,0.4mT工頻磁場分別暴露5min,15min,30min和1hour后,用改良的Folch法進(jìn)行脂類提取,采用高效液相色譜串聯(lián)質(zhì)譜法PIS模式下分析FL細(xì)胞內(nèi)神經(jīng)酰胺的種類,在MRM模式下分析不同神經(jīng)酰胺含量變化；隨后分別用伏馬菌素(FB1)和丙咪嗪(Imipramine)預(yù)處理抑制神經(jīng)酰胺從頭合成途徑和酸性鞘磷脂酶水解途徑,用N-acetylcysteine(NAC)預(yù)處理抑制活性氧(ROS)后,以工頻磁場暴露15min,并采用同樣的方法提取并檢測FL細(xì)胞內(nèi)神經(jīng)酰胺含量的變化。實(shí)驗(yàn)結(jié)果用SPSS16.0統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì)分析。 實(shí)驗(yàn)結(jié)果： 經(jīng)工頻磁場輻照5min,15min,30min和1hour后,細(xì)胞內(nèi)神經(jīng)酰胺含量明顯升高,且C16CER均有統(tǒng)計(jì)學(xué)意義(1hour組P0.01,其余組P＜0.05),C18CER在5min差異有統(tǒng)計(jì)學(xué)意義(P0.05)；C24CER在15min(P0.01)和1hour(P0.05)差異有統(tǒng)計(jì)學(xué)意義。伏馬菌素(FB1)預(yù)處理可抑制工頻磁場暴露對神經(jīng)酰胺含量的誘導(dǎo)作用,然而沒有統(tǒng)計(jì)學(xué)差異；而丙咪嗪(Imipramine)預(yù)處理則能顯著抑制工頻磁場對C24CER的誘導(dǎo)作用(P0.05),C18CEP啥量雖有有一定程度降低,但未見有統(tǒng)計(jì)學(xué)差異。NAC預(yù)處理同樣可以顯著抑制工頻磁場對C24CER神經(jīng)酰胺的誘導(dǎo)作用(P0.05)。 結(jié)論： 1.50Hz工頻磁場暴露可以誘導(dǎo)FL細(xì)胞內(nèi)C16,C18和C24神經(jīng)酰胺含量的增高。 2.工頻磁場暴露誘導(dǎo)FL細(xì)胞內(nèi)神經(jīng)酰胺含量的增高可能通過酸性鞘磷脂酶的水解途徑和從頭合成途徑。 3.工頻磁場誘導(dǎo)神經(jīng)酰胺含量(C24CER)的升高與細(xì)胞內(nèi)ROS存在一定相關(guān)性。
[Abstract]:Objective: to study the effect of power frequency magnetic field exposure on ceramide metabolism in FL cells. Methods: human amniotic membrane (FL) cells were exposed to 50Hz and 0.4mT power frequency magnetic field for 5 min. After 15 min, 30 min and 1hour, the lipids were extracted by modified Folch method. The types of ceramide in FL cells were analyzed by high performance liquid chromatography (HPLC) tandem mass spectrometry (PIS), and the changes of ceramide content in FL cells were analyzed under MRM mode. Subsequently, fumonisin (FB1) and imipramine (Imipramine) pretreatment were used to inhibit ceramide ab initio synthesis pathway and acid sphingomyelase hydrolysis pathway, respectively. After N-acetylcysteine (NAC) pretreatment, reactive oxygen species (ROS) was suppressed and exposed to power frequency magnetic field for 15 min. The changes of ceramide content in FL cells were extracted and detected by the same method. The experimental results were analyzed by SPSS16.0 statistical software. The results showed that after 5 min of power frequency magnetic field irradiation, the content of ceramide in the cells increased significantly at 30 min, 30 min and 1hour, and the content of C16CER was significantly higher than that in the other groups (P0.01 in 1hour group and P < 0.05 in other groups). The difference of C18CER in 5min was statistically significant (P 0.05). There was significant difference in C24CER between 15min (P 0.01) and 1hour (P 0.05). Fumonisin (FB1) pretreatment could inhibit the induction of ceramide content induced by power frequency magnetic field exposure, but there was no significant difference. Imipramine (Imipramine) pretreatment could significantly inhibit the induction of C24CER by power frequency magnetic field (P 0.05). Although the amount of C18CEP decreased to a certain extent, However, there was no significant difference. NAC pretreatment could also significantly inhibit the induction of C24CER ceramide by power frequency magnetic field (P 0.05). Conclusion: 1.50Hz power frequency magnetic field exposure can induce the increase of C 16, C 18 and C 24 ceramide contents in FL cells. two銆，

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