电针大脑中动脉闭塞模型大鼠血清减轻体外新生乳鼠神经细胞的缺糖低氧损伤的作用研究

目的 观察"电针血清"对新生乳鼠皮质神经细胞糖氧剥夺/复氧(oxygen-glucose depriva-tion and reoxygenation,OGD/R)损伤的作用.方法 将成年SD大鼠随机分为对照组、模型组、电针组,采用电凝法复制大脑中动脉闭塞模型;7d后对各组大鼠进行腹主动脉取血,离心,抽取上清,分离新生...

Effect of Electroacupuncture Serum in Alleviating Oxygen-Glucose Deprivation/Reoxygenation Injury of Neural Cells from Neonatal Mice in vitro in a Rat Model of Middle Cerebral Artery Occlusion

Objective To investigate the effect of electroacupuncture serum on oxygen-glucose deprivation/reoxygenation(OGD/R)injury of cortical neurons from neonatal mice.Methods Adult Sprague-Dawley(SD)rats were randomly divided into control group,model group,and electroacupuncture group.A rat model of middle cerebral artery occlusion model(MCAO)was established by electrocoagulation;at 7 days after modeling,blood was collected from the abdominal aorta and supernatant was extracted by centrifugation.Cortical neurons were isolated from neonatal SD rats within 24 hours after birth and were then randomly divided into control group,model group,and electroacupuncture serum group.The neurons in the control group were cultured in the medium of normal rat serum,those in the model group were treated with oxygen-glucose deprivation for 2 hours followed by glucose and oxygenation for 24 hours,and those in the electroacupuncture serum group were cultured in the medium containing electroacupuncture serum for 24 hours,followed by the same treatment as the model group.MTT assay was used to measure the viability of neurons;TUNEL method was used to measure neuronal apoptosis;the lactate dehydrogenase(LDH)release assay was used to measure the level of LDH leakage in neuronal culture medium;the kit was used to measure the levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in neurons;Western blot was used to measure the protein expression levels of Wnt-1,glycogen synthase kinase-3β(GSK-3β),β-catenin,neuronal nuclear protein(NeuN)in neurons.Results Compared with the control group,the model group had a significant reduction in the viability of neurons(P<0.05)and a significant increase in the apoptosis rate of neurons(P<0.05);compared with the model group,the electroacupuncture serum group had a significant increase in the viability of neurons(P<0.05)and a significant reduction in the apoptosis rate of neurons(P<0.05).Compared with the control group,the model group had a significant reduction in SOD activity(P<0.05)and significant increases in MDA content and LDH leakage(P<0.05);compared with the model group,the electroacupuncture serum group had a significant increase in SOD activity(P<0.05)and significant reductions in MDA content and LDH leakage(P<0.05).Compared with the control group,the model group had significant reductions in the protein expression levels of Wnt-1,GSK-3β,β-catenin,and NeuN(P<0.05);compared with the model group,the electroacupuncture serum group had significant increases in the protein expression levels of Wnt-1,β-catenin,and NeuN(P<0.05)and a significant reduction in the protein expression level of GSK-3β(P<0.05).Conclusion Electroacupuncture serum can alleviate the injury of cortical neurons induced by hypoxia and glucose deprivation and reduce the apoptosis of neurons,possibly by activating the Wnt/β-catenin signaling pathway.