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目的 探讨胆红素诱导原代培养神经细胞内Ca2 + 含量的变化 ,以及MgSO4 的拮抗作用。方法 采用胎鼠大脑皮层细胞作原代神经细胞培养 ,观察 10 0 μmol/L胆红素对不同成熟程度神经细胞内Ca2 + 含量的影响。原代培养神经细胞经Fura- 2 /AM负载 ,荧光图像分析测定细胞内Ca2 + 含量。结果 未成熟神经细胞 (培养2天 )暴露于 10 0 μmol/L胆红素 4h ,细胞内Ca2 + 从 75 .3nmol增加到 32 0 .8nmol;继续暴露 4h ,细胞内Ca2 + 增加到 40 0 .7nmol;去除胆红素暴露 ,细胞内Ca2 + 不能逆转。成熟神经细胞 (培养 8天 )暴露于 10 0 μmol/L胆红素 4h ,细胞内Ca2 + 从 70 .6nmol增加到 15 0 .8nmol;继续暴露 4h ,细胞内Ca2 + 不再增加 ;去除胆红素暴露 ,细胞内Ca2 + 浓度可显著降低。培养液中Mg2 + 加至 2mmol/L可减轻胆红素诱导的神经细胞内Ca2 + 含量变化。结论 胆红素可诱导原代培养的、未成熟的神经细胞Ca2 + 超载 ,MgSO4 可拮抗胆红素诱导的神经细胞Ca2 + 超载。
Objective To investigate the change of intracellular Ca2 + content induced by bilirubin in primary cultured neurons and the antagonism of MgSO4. Methods Fetal cortex cells were cultured as primary neurons to observe the effects of 100 μmol / L bilirubin on Ca2 + content in neurons of different maturation levels. Primary cultured neurons were loaded with Fura-2 / AM and intracellular Ca2 + contents were determined by fluorescence image analysis. Results Immature neurons (cultured for 2 days) were exposed to 10 μmol / L bilirubin for 4 hours, intracellular Ca2 + increased from 75.3 nmol to 32.08 nmol, and continued to be exposed for 4 hours. The intracellular Ca2 + increased to 40 °. 7nmol; Remove bilirubin exposure, intracellular Ca2 + can not be reversed. Mature neurons (cultured for 8 days) were exposed to 100 μmol / L bilirubin for 4 h, intracellular Ca2 + increased from 70.6 nmol to 150.8 nmol, and continued to be exposed for 4 h without increasing Ca2 + Su-exposed, intracellular Ca2 concentration can be significantly reduced. The addition of Mg2 + in the culture medium to 2 mmol / L can reduce the bilirubin-induced changes of Ca2 + in neurons. Conclusion Bilirubin can induce primary cultured and immature neurons to overload Ca2 +, and MgSO4 can antagonize bilirubin-induced Ca2 + overload in neurons.