摘要
目的探讨神经生长因子(NGF)改善脑组织低氧缺血状态作用机制。方法本研究通过培养混合神经干细胞(NSC)衍生的少突胶质细胞前体细胞(OPC)/星形胶质细胞培养来阐明神经生长因子在整个少突胶质细胞(OL)分化过程中的作用,并探讨其在缺血低氧状态条件下对OPC的保护作用。结果共聚焦显微镜检查结果显示,GW-44175610μM处理组细胞凝聚核百分比显著高于0.1μM、1μM及NGF抗体组(P<0.05)。抗NGF抗体处理组及GW-4417561μM处理组分化末期OPC百分比显著高于其他组(P<0.05)。抗NGF抗体处理组及GW-44175610μM处理组成熟OL及成髓鞘OPC百分比显著低于其他组(P<0.05)。抗NGF抗体处理组及GW-44175处理组NG2-阳性细胞比例显著高于其他组(P<0.05);抗NGF抗体处理组及GW-44175处理组CNPase/MBP染色细胞比例显著低于其他组(P<0.05);GW-441756处理组GFAP染色细胞比例显著高于其他组(P<0.05)。共聚焦显微镜检查结果显示,OGD处理组星形胶质细胞NGF mRNA表达水平显著高于无OGD处理组(P<0.05);NGF处理组细胞核内蛋白激酶B(AKT)荧光强度水平显著高于无NGF处理组(P<0.05);NGF处理组细胞核中磷酸化AKT荧光强度显著高于无NGF处理组、神经生长因子抗体(Ab-NGF)处理组及原肌球蛋白受体激酶A(TRKA)拮抗剂组(P<0.05)。结论NGF在体外细胞培养中与星形胶质细胞的相互作用可调节OPC生理分化;同时在缺血低氧状态下其对于OPC存活及OL成熟具有明确保护作用。
Objective To explore the mechanism of nerve growth factor in improving hypoxic and ischemic state of brain tissue.Methods In this study oligodendrocyte precursor cells(OPC)/astrocytes derived from mixed neural stem cells(NSC)were cultured to elucidate the role of nerve growth factor in the whole process of oligodendrocyte(OL)differentiation,and to explore its protective effect on oligodendrocyte precursor cells under the condition of ischemia and hypoxia.Results Confocal microscopy showed that the percentage of agglomerated nuclei in GW-44175610μM treatment group was significantly higher than that in 0.1μM,1μM and anti-NGF antibody groups(P<0.05).The percentage of OPC at the end of differentiation in anti-NGF antibody treatment group and GW-4417561μM treatment group was significantly higher than that in other groups(P<0.05).The percentages of mature OL and myelinated OPC in the anti-NGF antibody treatment group and GW-44175610μM treatment group were significantly lower than those in the other groups(P<0.05).The proportion of NG2-positive cells in anti-NG F antibody treatment group and GW-44175 treatment group was significantly higher than that in other groups(P<0.05).The proportion of CNPase/MBP staining cells in anti-NGF antibody treatment group and GW-44175 treatment group was significantly lower than that in other groups(P<0.05).The proportion of GFAP stained cells in GW-441756treatment group was significantly higher than that in other groups(P<0.05).Confocal microscopy showed that the NGF mRNA expression level of astrocytes treated with OGD was significantly higher than that without OGD(P<0.05).The fluorescence intensity of AKT in nucleus of NGF treated group was significantly higher than that of non-NGF treated group(P<0.05).The fluorescence intensity of phosphorylated AKT in the nucleus of NGF treated group was significantly higher than that of non-NGF treated group,Ab-NGF treated group and TRKA antagonist group(P<0.05).Conclusion The interaction of nerve growth factor with astrocytes in vitro cell cul
作者
刘萍萍
何学芳
张翼
杨旭
张珊珊
季一飞
Liu Pingping;He Xuefang;Zhang Yi;Yang Xu;Zhang Shanshan;Ji Yifei(Department of Neurology,Nanchong Central Hospital,Beijing Anzhen Hospital Nanchong Hospital,&Sichuan Provincial Clinical Medical Research Center for Neurological Diseases,Sichuan 637000,China)
出处
《脑与神经疾病杂志》
CAS
2024年第8期467-473,共7页
Journal of Brain and Nervous Diseases
基金
国家自然科学基金资助项目(81870966)
四川省自然科学基金项目(2022NSFSC0756)。
关键词
神经生长因子
少突胶质前体细胞
缺血低氧
发育性髓鞘
Nerve growth factor
Oligodendrocyte progenitor cells
Hypoxia/ischemia
Developmental myelin sheath
