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目的建立1,2-二氯乙烷(1,2-DCE)对离体培养的人神经母细胞瘤细胞系SH-SY5Y细胞的染毒模型,探讨1,2-DCE对SH-SY5Y细胞的毒性作用及机制。方法取对数生长期的SH-SY5Y细胞,分别以终浓度为0、10、20、30、40、50、60、70、80 mmol/L的1,2-DCE染毒并培养24 h后,观察细胞形态,以CCK-8法检测细胞存活率,采用化学比色法检测细胞培养上清液中乳酸脱氢酶(LDH)活力,细胞内丙二醛(MDA)水平和超氧化物歧化酶(SOD)、三磷酸腺苷(ATP)酶的活力。结果随着1,2-DCE染毒剂量的增加,SH-SY5Y细胞细胞密度逐渐降低,突触变短,胞膜破裂,胞质浓缩,胞质内容物外溢逐渐增多。随着1,2-DCE染毒剂量的增加,细胞存活率呈下降趋势(P<0.01),细胞培养上清液中LDH活力呈升高趋势(P<0.01),均呈剂量-效应关系;而细胞内MDA水平和SOD、钠-钾ATP酶、钙-镁ATP酶、总ATP酶的活力总体上均呈先升高后下降的趋势。细胞培养上清液中LDH活力与细胞存活率呈负相关(相关系数为-0.907,P<0.01)。结论 1,2-DCE对SH-SY5Y细胞有增殖抑制作用,其机制可能与1,2-DCE导致的细胞膜通透性改变,自由基对细胞损害加强,细胞对自由基清除能力的下降,ATP酶活力下降以及钙超载有关。SH-SY5Y细胞可作为1,2-DCE细胞毒性分析的常用细胞系。
Objective To establish a model of human neuroblastoma cell line SH-SY5Y cultured with 1,2-dichloroethane (1,2-DCE) and investigate the effect of 1,2-DCE on SH-SY5Y cells Toxicity and mechanism. Methods SH-SY5Y cells in logarithmic growth phase were treated with 1,2-DCE at final concentration of 0, 10, 20, 30, 40, 50, 60, 70 and 80 mmol / L for 24 h Cell morphology was observed. The cell viability was detected by CCK-8 assay. The activity of lactate dehydrogenase (LDH), intracellular malondialdehyde (MDA) and superoxide dismutase Enzyme (SOD), adenosine triphosphate (ATP) enzyme activity. Results With the increase of the dose of 1,2-DCE, the cell density of SH-SY5Y cells gradually decreased, the synapses became shorter, the membrane ruptured, the cytoplasm was concentrated and the content of cytoplasm increased gradually. With the increase of 1,2-DCE dose, the cell viability decreased (P <0.01) and the LDH activity in cell culture supernatant increased (P <0.01), showing a dose-response relationship. However, the intracellular MDA level and the activities of SOD, sodium-potassium ATPase, calcium-magnesium ATPase and total ATPase increased first and then decreased. LDH activity in cell culture supernatant was negatively correlated with cell viability (correlation coefficient -0.907, P <0.01). Conclusions 1,2-DCE can inhibit the proliferation of SH-SY5Y cells. The mechanism may be related to the change of cell membrane permeability induced by 1,2-DCE, the damage of free radicals to cells and the decrease of free radical scavenging capacity of ATP. Decreased enzyme activity and calcium overload. SH-SY5Y cells can be used as a common cell line for 1,2-DCE cytotoxicity assays.