蝎毒纤溶活性肽对低糖低氧/复氧损伤大鼠皮层神经元的保护作用

Protective effect of scorpion venom fibrinolytic active peptideson anoxia/reoxygenation injury of cortex neurons

目的:研究蝎毒纤溶活性肽(SVFAPs)对大鼠神经元低糖低氧/复氧损伤的影响,探讨药物发挥神经保护作用的可能机制。方法:建立原代培养的大鼠皮层神经元的低糖低氧/复氧(OGD/R)模型,采用四唑蓝(MTT)法和乳酸脱氢酶(LDH)释放法检测细胞活力;检测细胞上清液中丙二醛(MDA)和一氧化氮(NO)含量、超氧化物歧化酶(SOD)活性;Hoechst染色和DNA片段法检测细胞凋亡;W estern b lot法检测PARP蛋白的表达。结果:经OGD/R损伤后细胞活力明显下降,细胞上清液中MDA、NO升高,SOD降低;出现凋亡典型的形态学特征,PARP蛋白表达下降;蝎毒纤溶活性肽(2～8mg/L)可不同程度提高低糖低氧/复氧后神经元的活力,降低细胞上清液MDA和NO含量,提高SOD活性。改善凋亡相关的细胞核形态学改变,并且减少凋亡特征性DNA片段化的发生,降低细胞凋亡率,保持PARP的完整性。结论:蝎毒纤溶活性肽对低糖低氧/复氧损伤的神经元具有保护作用,其机制可能与抗氧化和减轻细胞凋亡有关。

Objective： To study the effect of scorpion venom fibrinolytic active peptides（SVFAPs）on the oxygen-glucose deprivation /reoxygenation（OGD/R）injured neuron in rat.Methods： Primary cultured rat＇s cortex neurons were made into OGD/R model cells.The cell viability was detected by MTT assay and lactate dehydrogenase releasing methods;the activity of superoxide dismutase（SOD）,and contents of malondialdehyde（MDA） and nitride oxide（NO） in culture supernate were detected;the apoptosis rate of cortex neurons after OGD/R was measured by hoechst33342 staining and DNA ladder;Western blot was used to detect the protein expression of PARP.Results ： SVFAPs enhanced the cell viability of neurons after OGD/R injury,increased SOD activity and decreased the MDA and NO contents in supernate,reduced the OGD/R-induced apoptosis and keep the instate of PARP.Conclusion： SVFAPs has the protective effects on OGD/R injured neurons,the mechanism is possibly related with its anti-oxidation and induced apoptosis.