乙醇、乙醛对原代培养星形胶质细胞神经甾体合成的影响

 目的研究长时程乙醇、乙醛处理对原代培养大鼠大脑皮质星形胶质细胞神经甾体合成的影响。方法采用MTT法测定不同浓度乙醇、乙醛对星形胶质细胞活性和存活率的影响,并采用HPLC-MS分离测定游离型神经甾体孕烯醇酮(PREG)、脱氢表雄酮(DHEA)、别孕烯醇酮(AP)及结合型神经甾体脱氢表雄酮硫酸酯(DHEAS)、孕烯醇酮硫酸酯(PREGS)的含量,观察不同浓度乙醇、乙醛处理72 h后,原代培养的大鼠大脑皮质神经星形胶质细胞中各类神经甾体合成释放的变化。结果不同浓度乙醇、乙醛处理72 h后,各处理组星形胶质细胞活性及存活率与对照组相比无显著性差异。与PBS对照组比较,低浓度(25 mmol·L^-1)和中浓度(50 mmol·L^-1)乙醇分别使DHEAS和DHEA水平显著下降(P<0.05);中、高(100 mmol·L^-1)浓度乙醇可使AP水平显著升高(P<0.05,P<0.001)、PREGS水平显著下降(P<0.05),高浓度乙醇还可显著降低PREG的水平(P<0.05);低浓度(12.5 mmol·L^-1)乙醛可使PREG水平显著降低(P<0.05),而中(25 mmol·L^-1)、高(50 mmol·L^-1)浓度乙醛可使DHEA水平显著增高(P<0.05)。结论乙醇和乙醛对神经星形胶质细胞神经甾体的合成均有一定影响,但二者的作用特点有所不同。

Effect of Ethanol and Aldehyde on Levels of Neurosteroids in Primary Cultural Astrocytes

OBJECTIVE To evaluate the effect of long term treatment of ethanol and aldehyde on levels of neurosteroids in primary cultural cortex astrocytes from fetal rats brain. METHODS The influence of ethanol and aldehyde on the livability and survival rate of primary cultural cortex astrocytes was evaluated by MTT.The concentration of unconjugated neurosteroids(PREG,DHEA and AP) and conjugated neurosteroids(DHEAS and PREGS)in Media from primary cultural cortex astrocyte were analyzed and determined using HPLC-MS/APCI.The effect on levels of neurosteroids after 72 h treatment with differents concentrations ethanol and aldehyde was evaluated. RESULTS No Significant effect of 12.5～100 mmol·L^-1ethanol and 6.25～50 mmol·L^-1aldehyde on the livability and survival rate of primary cultural cortex astrocytes was abserved.After 72 h treatment with three concentrations of ethanol and aldehyde,the levels of DHEAS after treatment with 25 mmol·L^-1 ethanol and the level of DHEA after treatment with 50 mmol·L^-1 ethanol were significant decreased(P<0.05).The level of AP was significant increased(P<0.05,P<0.001),but PREGS was significant decreased(P<0.05) after treatment with 50 mmol·L^-1 or 100 mmol·L^-1 ethanol.The level of PREG was significant decreased(P<0.05) after treatment with 100 mmol·L ethanol and 12.5 mmol·L^-1 aldehyde,The level of DHEA was significant increased(P<0.05) after treatment with 25 and 50 mmol·L^-1 aldehyde.Other treatment have no obviously effects on unconjugated and conjugated neurosteroids compared with PBS control group.CONCLUSION Both ethanol and aldehyde show effects on levels of neurosteroids in primary cultural cortex astrocytes with different characteristics.