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目的探讨对氨基水杨酸钠(PAS-Na)对体外染锰致大鼠原代基底核神经元损伤的影响。方法培养至第8天的基底核神经元随机分为正常对照组(对照组)、低、中、高剂量染锰(L-、M-、H-Mn)组、PAS-Na(PAS)对照组、低、中、高剂量PAS-Na(L-、M-、H-PAS)干预组。对照组神经元给予培养液培养24 h;染锰组神经元暴露于MnCl2.4H2O 100、200和400μmol/L培养液培养24 h;PAS对照组神经元给予含PAS-Na 50、500和5000μmol/L培养液培养24 h。L-、M-、H-PAS干预组神经元分别暴露于MnCl2.4H2O 100、200和400μmol/L培养液培养24 h,接着弃掉原培养液,再分别给予含PAS-Na 50、500和5000μmol/L的培养液培养24 h。其余组培养液培养24 h。然后,用噻唑蓝(MTT)法测定存活率,单细胞凝胶电泳技术(SCGE)测定DNA损伤,硫代巴比妥酸(TBA)法测定丙二醛(MDA)。结果与对照组相比,染锰组神经元存活率明显降低,彗星尾部DNA百分率、Olive尾距增高,MDA含量增加,差异有统计学意义(P<0.05)。L-、M-PAS干预使暴露于L-Mn组神经元存活率提高,彗星尾部DNA百分率、Olive尾距降低,MDA含量减少;M-PAS干预使暴露于M-Mn组神经元存活率提高,彗星尾部DNA百分率、Olive尾距降低,差异有统计学意义(P<0.05)。结论体外染锰对大鼠原代基底核神经元损伤明显,PAS-Na对锰致基底核神经元毒性有一定的干预作用。
Objective To investigate the effects of sodium salicylate (PAS-Na) on primary cultured rat primary basal ganglia neurons injured by manganese. Methods The basal ganglia neurons cultured on the 8th day were randomly divided into normal control group (control group), low, middle and high doses of manganese (L-, M-, H-Mn) group, PAS-Na Group, low, medium and high doses of PAS-Na (L-, M-, H-PAS) intervention group. The neurons in the control group were cultured for 24 h. The neurons in the manganese-exposed group were exposed to MnCl2.4H2O at 100, 200 and 400 μmol / L for 24 h. The neurons in the PAS control group were treated with PAS-Na at concentrations of 50, 500 and 5000 μmol / L culture medium for 24 h. Neurons exposed to L-, M- and H-PAS groups were incubated with MnCl2.4H2O at 100, 200 and 400 μmol / L for 24 h, then discarded the original culture medium, and then treated with PAS-Na 50,500 and 5000μmol / L culture medium for 24 h. The remaining group of culture medium for 24 h. Then, the survival rate was determined by MTT assay, DNA damage was determined by single cell gel electrophoresis (SCGE), and malondialdehyde (MDA) was measured by thiobarbituric acid (TBA). Results Compared with the control group, the survival rate of neurons in Mn-Mn group was significantly decreased. The tail DNA percentage, Olive tail distance and MDA content in comets were significantly increased (P <0.05). The survival rate of neurons exposed to L-Mn group increased, the DNA percentage of comet tail, Olive tail distance decreased and MDA content decreased after intervention with L- and M-PAS. The survival rate of neurons exposed to M-Mn increased , Tail DNA percentage, Olive tail distance decreased, the difference was statistically significant (P <0.05). Conclusion In vitro manganese exposure significantly damages neurons in the primary basal ganglia of rats, and PAS-Na may interfere with the neurotoxicity induced by manganese in basal ganglia.