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目的通过观察砷对星形胶质细胞(AST)分泌D-丝氨酸的影响,探讨砷的智力损伤作用机制。方法以原代培养脑AST为试验对象,在含砷浓度分别为0、2.5、5、10μmol/L的培养液中培养24 h,采用高效液相色谱法检测D-丝氨酸水平,Western blot法检测丝氨酸消旋酶(SR)蛋白表达水平,荧光双波长分光光度计法检测细胞内游离钙离子([Ca2+]i)浓度。结果与对照组比较,2.5、5、10μmol/L砷暴露后,AST分泌D-丝氨酸的水平[分别为(21.580±1.313)、(21.936±1.539)、(23.401±1.648)μmol/L]和SR蛋白表达水平[分别为(1.327±0.122)、(1.397±0.105)、(1.403±0.104)]均明显升高,10μmol/L砷暴露组AST内[Ca2+]i含量明显升高。结论砷暴露可引起AST分泌D-丝氨酸水平增加,该过程可能与砷暴露引起AST内SR的表达水平增强以及[Ca2+]i升高有关。
Objective To observe the effect of arsenic on the secretion of D-serine by astrocytes (AST) and to explore the mechanism of arsenic damage. Methods Primary cultured brain AST was cultured in culture media containing 0, 2.5, 5, 10 and 10 μmol / L arsenic concentrations for 24 h. The levels of D-serine were determined by high performance liquid chromatography (HPLC) Serine gyrase (SR) protein expression level, intracellular free calcium ion ([Ca2 +] i) concentration was measured by fluorescence double-wavelength spectrophotometry. Results Compared with the control group, the levels of D-serine secreted by AST after 2.5, 5, 10μmol / L arsenic exposure were 21.580 ± 1.313, 21.936 ± 1.539 and 23.401 ± 1.648 μmol / L, respectively (1.327 ± 0.122, 1.397 ± 0.105, 1.403 ± 0.104, respectively] were significantly higher than those in the arsenic-exposed group (P <0.05). The contents of [Ca2 +] i in the 10 μmol / L arsenic exposure group were significantly increased. Conclusion Arsenic exposure may induce the increase of D-serine secretion in AST, which may be related to the increased expression of SR and the increase of [Ca2 +] i in AST.