氯化锰致人骨髓神经母细胞瘤细胞株线粒体损伤及对多巴胺分泌和PARK2表达的影响
发布时间：2019-07-23 09:16
【摘要】：[目的]研究氯化锰对人骨髓神经母细胞瘤细胞株(SH-SY5Y)线粒体损伤、氧化应激、多巴胺分泌及PARK2表达的影响。[方法]0、100、300、500μmol/L浓度氯化锰染毒SH-SY5Y细胞24 h后,用MTT法测细胞抑制率(反映线粒体损伤情况),石墨炉原子吸收光谱法测定细胞内锰浓度,高度水溶性四唑盐(WST-1)法测定细胞内超氧化物歧化酶(SOD)活性,硫代巴比妥酸法测定细胞内丙二醛(MDA)含量,反相高效液相色谱-荧光法测定细胞内多巴胺(DA)含量,实时荧光定量-PCR检测PARK2 m RNA表达,蛋白免疫印迹法检测Parkin蛋白表达。[结果]与对照组比较,MnCl2浓度为300、500μmol/L时,细胞抑制率(线粒体损伤)增高(P0.01)。与对照组比较,染锰组细胞内锰浓度升高(P0.05或P0.01)。与对照组比较,MnCl2浓度为300、500μmol/L时,SOD活性和DA含量降低(P0.01),MDA含量升高(P0.01);细胞PARK2 m RNA表达和Parkin蛋白表达降低(P0.01)。相关性分析显示,PARK2 m RNA表达与细胞抑制率(线粒体损伤)、细胞内锰浓度及MDA含量呈负相关,r值分别为-0.872、-0.880、-0.862(均P0.01);PARK2 m RNA表达与SOD活性、DA含量以及Parkin蛋白表达呈正相关,r值分别为0.879、0.859、0.809(均P0.01)。[结论]氯化锰暴露可引起SH-SY5Y细胞的线粒体损伤、氧化应激、DA分泌减少和PARK2表达下降。
[Abstract]:[objective] to study the effects of manganese chloride on mitochondrial damage, oxidative stress, dopamine secretion and PARK2 expression in human bone marrow neuroblastoma cell line (SH-SY5Y). [methods] SH-SY5Y cells were exposed to manganese chloride at the concentration of 0100300500 渭 mol / L for 24 h. The cell inhibition rate was measured by MTT, the intracellular manganese concentration was measured by graphite furnace atomic absorption spectrometry (GFAAS), the intracellular SOD (SOD) activity was measured by highly water-soluble tetrazolium salt (WST-1) method, and the intracellular malondialdehyde (MDA) (MDA) content was measured by thiobarbituric acid method. The content of dopamine (DA) in cells was determined by reversed-phase high performance liquid chromatography-fluorescence method, the expression of PARK2 m RNA was detected by real-time fluorescence quantitative PCR, and the expression of Parkin protein was detected by Western imprinting. [results] compared with the control group, when the concentration of MnCl2 was 300500 渭 mol / L, the cell inhibition rate (mtDNA damage) was increased (P 0.01). Compared with the control group, the intracellular manganese concentration in the manganese exposed group was higher than that in the control group (P 0.05 or P 0.01). Compared with the control group, when the concentration of MnCl2 was 300500 渭 mol / L, the activity of SOD and the content of DA decreased (P 0.01), MDA), and the expression of PARK2 m RNA and Parkin protein decreased (P 0.01). Correlation analysis showed that the expression of PARK2 m RNA was negatively correlated with cell inhibition rate (mtDNA damage), intracellular manganese concentration and MDA content, r value was-0.872, 鈮，
本文编号：2518056
[Abstract]:[objective] to study the effects of manganese chloride on mitochondrial damage, oxidative stress, dopamine secretion and PARK2 expression in human bone marrow neuroblastoma cell line (SH-SY5Y). [methods] SH-SY5Y cells were exposed to manganese chloride at the concentration of 0100300500 渭 mol / L for 24 h. The cell inhibition rate was measured by MTT, the intracellular manganese concentration was measured by graphite furnace atomic absorption spectrometry (GFAAS), the intracellular SOD (SOD) activity was measured by highly water-soluble tetrazolium salt (WST-1) method, and the intracellular malondialdehyde (MDA) (MDA) content was measured by thiobarbituric acid method. The content of dopamine (DA) in cells was determined by reversed-phase high performance liquid chromatography-fluorescence method, the expression of PARK2 m RNA was detected by real-time fluorescence quantitative PCR, and the expression of Parkin protein was detected by Western imprinting. [results] compared with the control group, when the concentration of MnCl2 was 300500 渭 mol / L, the cell inhibition rate (mtDNA damage) was increased (P 0.01). Compared with the control group, the intracellular manganese concentration in the manganese exposed group was higher than that in the control group (P 0.05 or P 0.01). Compared with the control group, when the concentration of MnCl2 was 300500 渭 mol / L, the activity of SOD and the content of DA decreased (P 0.01), MDA), and the expression of PARK2 m RNA and Parkin protein decreased (P 0.01). Correlation analysis showed that the expression of PARK2 m RNA was negatively correlated with cell inhibition rate (mtDNA damage), intracellular manganese concentration and MDA content, r value was-0.872, 鈮，
本文编号：2518056
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