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目的研究不同浓度的氯化镧对原代大鼠星形胶质细胞内钙离子浓度、线粒体膜电位、线粒体及胞浆中细胞色素C的影响,探讨氯化镧所产生神经毒性的机制。方法取新生大鼠大脑皮质进行原代星形胶质细胞分离和培养。在纯化及鉴定细胞后,用0、0.25、0.5、1.0mmol/L氯化镧(LaCl3)染毒24h后,应用四甲基偶氮唑盐法检测细胞活力,以荧光探针法测定细胞内钙([Ca2+]i)的变化,以流式细胞仪法测定线粒体膜电位,以化学比色法测定线粒体和胞浆中细胞色素C含量。结果与对照组相比,随着氯化镧暴露浓度的增加,细胞存活率从100%逐渐下降至53.31%,有统计学意义(P<0.01),细胞内[Ca2+]i从(129.45±1.09)nmol/L逐渐升高至(224.84±5.51)nmol/L,有统计学意义(P<0.05),细胞线粒体膜电位从(159.16±4.76)逐渐降低至(85.70±5.94),有统计学意义(P<0.05),细胞线粒体中细胞色素C含量从(3.71±0.33)μmol/mg Pro逐渐降低至(2.55±0.08)μmol/mg Pro,有统计学意义(P<0.05),胞浆中细胞色素C含量从(0.31±0.19)μmol/mg Pro逐渐升高至(1.72±0.31)μmol/mg Pro,有统计学意义(P<0.05),并呈剂量-效应关系。结论氯化镧可致原代大鼠星形胶质细胞内钙稳态失衡,线粒体功能障碍,从而产生神经毒性。
Objective To study the effects of lanthanum chloride (Lanthanum chloride) on the concentration of intracellular calcium, mitochondrial membrane potential, cytochrome C in mitochondria and cytoplasm of primary rat astrocytes, and to explore the mechanism of lanthanum chloride-induced neurotoxicity. Methods The primary astrocytes were isolated and cultured in the cerebral cortex of neonatal rats. After purification and identification of the cells, the cells were treated with 0, 0.25, 0.5 and 1.0 mmol / L LaCl3 for 24 h, and the cell viability was measured by MTT assay. The change of calcium ([Ca2 +] i) was measured by flow cytometry. The contents of cytochrome C in mitochondria and cytoplasm were determined by chemical colorimetry. Results Compared with the control group, the cell viability decreased from 100% to 53.31% with the increase of the exposure concentration of lanthanum chloride (P <0.01), and the intracellular [Ca2 +] i increased from (129.45 ± 1.09) ) was increased to (224.84 ± 5.51) nmol / L with statistical significance (P <0.05). The mitochondrial membrane potential decreased from (159.16 ± 4.76) to (85.70 ± 5.94), with statistical significance (P <0.05). The content of cytochrome C in mitochondria decreased from (3.71 ± 0.33) μmol / mg Pro to (2.55 ± 0.08) μmol / mg Pro in mitochondria (P <0.05) Pigment C content increased from (0.31 ± 0.19) μmol / mg Pro to (1.72 ± 0.31) μmol / mg Pro, statistically significant (P <0.05), and showed a dose-response relationship. Conclusion Lanthanum chloride can cause the imbalance of calcium homeostasis and mitochondrial dysfunction in primary rat astrocytes, resulting in neurotoxicity.