2,3-吲哚醌对多巴胺能细胞氧化损伤的保护作用

目的采用过氧化氢(H2O2)作为损伤因素,检测2,3-吲哚醌(isatin,ISA)对MES 23.5细胞的保护作用及其机制。方法 MTT比色法检测H2O2的毒性作用及ISA的保护作用。细胞分为:对照组,H2O2 20μmol.L-1组,和ISA 100μmol.L-1+H2O2 20μmol.L-1组,采用流式细胞技术(FCM)检测线粒体膜电位(△ψm)的变化;激光扫描共聚焦显微镜(LSCM)检测细胞内Ca2+浓度(Ca2+]i)变化。结果 H2O2组细胞存活率明细降低。ISA 100μmol.L-1及以上浓度处理的细胞存活率均高于H2O2组(P<0.05)。H2O2组细胞内R123平均荧光强度较对照组明显降低而ISA组明显增强(P<0.01)。H2O2组细胞内Fluo-3荧光强度明显高于对照组(P<0.01),而ISA组细胞内荧光强度明显低于H2O2组(P<0.01)。结论 ISA可减轻H2O2导致的DA能MES 23.5细胞损伤,其机制与减轻△ψm异常,降低Ca2+]i水平有关。

Protective effects of isatin on H2O2-damaged dopaminergic cells

Aim MES 23.5 was incubated with different concentrations of H2O2 and H2O2 plus isatin(ISA),to detect the protective effect and its mechanism of ISA on MES 23.5 cells.Methods The MTT assay was used to detect the cell viability;FCM was used to detect the the mitochondrial membrane potential(△ψm).The changes of intracellular free calcium concentration(i)were monitored by laser scanning confocal microscope(LSCM) using Fluo-3/AM staining.Results H2O2 decreased the viability of MES 23.5 cells dose-dependently(P<0.01).The viability of MES 23.5 cells incubated with 20 μmol·L-1 H2O2 plus different concentration of ISA was increased compared with H2O2 treated alone(P<0.05).The result of FCM showed the average fluorescence intensity of R 123 in H2O2-treated cells was lower than that in control(P<0.01).The average fluorescence intensity in ISA 100 μmol·L-1 treated cells was higher than that in H2O2 group(P<0.01).The result of LSCM showed the average fluorescence intensity of Fluo-3 in H2O2 incubation cells was higer than that in control group(P<0.01);in ISA pretreatment group,the average fluorescence intensity was lower than that in H2O2 treated alone group(P<0.01).Conclusion ISA protects against the damage of H2O2 on MES 23.5 cells,mitigating dysfunction of △ψm,and decreasing the i are the underlying mechanisms of ISA protective effect.