發(fā)布時間：2019-07-23 09:16

【摘要】：[目的]研究氯化錳對人骨髓神經(jīng)母細(xì)胞瘤細(xì)胞株(SH-SY5Y)線粒體損傷、氧化應(yīng)激、多巴胺分泌及PARK2表達(dá)的影響。[方法]0、100、300、500μmol/L濃度氯化錳染毒SH-SY5Y細(xì)胞24 h后,用MTT法測細(xì)胞抑制率(反映線粒體損傷情況),石墨爐原子吸收光譜法測定細(xì)胞內(nèi)錳濃度,高度水溶性四唑鹽(WST-1)法測定細(xì)胞內(nèi)超氧化物歧化酶(SOD)活性,硫代巴比妥酸法測定細(xì)胞內(nèi)丙二醛(MDA)含量,反相高效液相色譜-熒光法測定細(xì)胞內(nèi)多巴胺(DA)含量,實時熒光定量-PCR檢測PARK2 m RNA表達(dá),蛋白免疫印跡法檢測Parkin蛋白表達(dá)。[結(jié)果]與對照組比較,MnCl2濃度為300、500μmol/L時,細(xì)胞抑制率(線粒體損傷)增高(P0.01)。與對照組比較,染錳組細(xì)胞內(nèi)錳濃度升高(P0.05或P0.01)。與對照組比較,MnCl2濃度為300、500μmol/L時,SOD活性和DA含量降低(P0.01),MDA含量升高(P0.01);細(xì)胞PARK2 m RNA表達(dá)和Parkin蛋白表達(dá)降低(P0.01)。相關(guān)性分析顯示,PARK2 m RNA表達(dá)與細(xì)胞抑制率(線粒體損傷)、細(xì)胞內(nèi)錳濃度及MDA含量呈負(fù)相關(guān),r值分別為-0.872、-0.880、-0.862(均P0.01);PARK2 m RNA表達(dá)與SOD活性、DA含量以及Parkin蛋白表達(dá)呈正相關(guān),r值分別為0.879、0.859、0.809(均P0.01)。[結(jié)論]氯化錳暴露可引起SH-SY5Y細(xì)胞的線粒體損傷、氧化應(yīng)激、DA分泌減少和PARK2表達(dá)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，

本文編號：2518056