锰对神经元细胞内钙稳态影响的研究

Study of the effects of manganese on intracellular Ca^(2+) homeostasis in primary cultured neurons

目的研究锰对神经细胞内钙稳态的影响,重点观察神经元细胞钙离子浓度、Na+-K+-ATPase和Ca2+-ATPase活性的改变。方法选用原代培养神经元为模型,待细胞生长至最佳状态时,予以分组处理:锰处理组为含不同浓度氯化锰(0,25,100,400μmol/L)的培养液,培养神经细胞12h后,通过倒置相差显微镜观察细胞形态的改变,并检测神经元细胞内钙离子浓度、Na+-K+-ATPase和Ca2+-ATPase活性的变化。结果随着染Mn剂量的加大,神经元形态出现异常,细胞内钙离子浓度逐渐升高;细胞Na+-K+-ATPase和Ca2+-ATPase活性也逐渐下降。结论锰通过影响与维持钙稳态相关的酶(Na+-K+-ATPase和Ca2+-ATPase)活性,干扰神经元细胞内钙稳态,进而造成神经元细胞损伤。

Objectives To explore the effect of manganese （Mn） on intracellular calcium concentration （[Ca2＋]i）, and activities of Na＋-K＋-ATPase and Ca2＋-ATPase in primary cultured neurons.Methods Primary cultured neurons was cultured with fresh 0,25,100,400 μmol/L, manganese chloride media for 12h, respectively; then observed morphological changes by inverted phase contrast microscope and detected [Ca2＋]i, and activities of Na＋-K＋-ATPase and Ca2＋-ATPase in primary cultured neurons.Results The results showed that the neurons were gradually shrinked, meanwhile, the neuraxons were shortened, and the networks among neurons were disappeared along with the increase of Mn concentration,[Ca2＋]i was increased. Additionally, the activities of Na＋-K＋-ATPase and Ca2＋-ATPase were significantly decreased in a concentration-dependent manner.Conclusions It was suggested that Mn could impair neurons by disturbing intracellular Ca2＋ homeostasis and inhibiting activity of Na＋-K＋-ATPase and Ca2＋-ATPase.