苦参素对脂多糖诱导的海马神经元凋亡的保护作用

Protective effects of oxymatrine on LPS-induced neuronal apoptosis of hippocampal neurons

目的探讨苦参素(OMT)对脂多糖(LPS)诱导的海马神经元凋亡的影响及其可能机制。方法根据海马神经元乳酸脱氢酶(LDH)释放率选择OMT的浓度进行实验分组。分为7组:正常对照组;脂多糖组;阿司匹林+脂多糖组;苦参素组;苦参素低剂量+脂多糖组;苦参素中剂量+脂多糖组;苦参素高剂量+脂多糖组。流式细胞仪检测细胞凋亡率;免疫荧光观察NF KB/P65核易位情况;酶联免疫吸附(ELISA)技术测定海马神经元培养上清中炎症因子肿瘤坏死因子-a(TNF-a)和白细胞介素-1β(IL-1β)的水平。结果LPS组海马神经元凋亡率显著高于正常对照组(P<0.01),OMT预处理可以显著减少LPS刺激引起的海马神经元凋亡率(P<0.05)。LPS可以促进海马神经元NF-kB/P65由胞浆转位到胞核.提高海马神经元培养上清中TNF-a和IL-1β水平;而OMT预处理能显著减少LPS对海马神经元NF-κB/P65核易位作用及降低TNF-a和IL-1β的水平(P<0.05)。结论LPS诱导海马神经元NF-κB/P65核易位,导致炎症因子TNF-a和IL-1β的含量增加;OMT预处理可以抑制海马神经元NF-κB/P65核易位,减少TNF-a和IL-1β的分泌,从而抑制海马神经元的凋亡。

Objective to investigate the ffeet and mechanisms of oxymatrine(OMT)on lipopolysaccharide(LPS)in-duced apoptosis of hippocampal neurons.Methods According to the release rate of lactate dehydrogenase(LDH)in hippocampal neurons,the concentration of OMT was selected for experimental grouping.There were seven groups:nor-mal group,LPS group,Aspirin+LPS group,OMT group,OMT low-dose+LPS group,OMT mid-dose+LPS group and OMT high-dose+LPS group.Apoptosis rate was detected by flow cytometry.The nuclear translocation of NF-KB/P65 was observed by immunofluorescence.The level of tumor necrosis factor-a(TNF-a)and interleukin-1β(IL-1β)in culture supernatants of hippocampal neurons were determined by Enzyme-linked immunosorbent assay(ELISA).Results The apoptosis rate of hippocampal neurons in the LPS group was higher than normal group(P<0.01),and OMT could sig-nificantly reduce LPS stimulated apoptosis rate of hippocampal neurons(P<0.05).LPS promoted NF-KB/P65 shifted from cytoplasm to the nucleus,and increased the level of TNF-a and IL-Iβ in the culture supernatant of hippocampal.neuron,while OMT reduced the LPS on hippocampal neurons NF-KB/P65 nuclear translocation and reduce the level of TNF-a and IL-1β.Conclusion LPS could induce NF-kB/P65 nuclear translocation in hippocampal neurons and result in increased levels of inflammatory factors TNF-a and IL-1β.OMT could weaken the nuclear translocation of NF-KB/P65 induced by LPS and reduce the level of TNF-a and IL-1β,which in turn to inhibit apoplosis of hippocampal neurons.