摘要
目的:探索程序性死亡分子1(PD1)敲除对小鼠脊髓损伤后巨噬/小胶质细胞极化、血管生成、神经元存活、空洞形成及运动功能恢复的效应,阐明PD1在脊髓继发性损伤中的作用,为进一步研发干预措施奠定基础。方法:采用PD1基因敲除小鼠,制备脊髓挫伤模型;免疫荧光化学染色研究巨噬/小胶质细胞极化、血管生成、神经元存活及空洞形成;实时荧光定量聚合酶链式反应(real time RT-PCR)和免疫印迹实验(Western Blot)研究炎症因子、血管生长因子的表达变化;Basso小鼠运动功能评分(BMS)评估小鼠的运动功能。结果:与野生型(WT)小鼠相比,PD1敲除小鼠脊髓损伤区M1型巨噬小胶质细胞相关分子诱导型一氧化氮合酶(iNOS)、白细胞分化抗原86(CD86)、白介素-1β(IL-1β)、白介素6(IL-6)和肿瘤坏死因子-α(TFN-α)的表达水平显著上调,M2型相关分子精氨酸酶1(Arg1)、白介素-10(IL-10)、白介素-4(IL-4)和转化生长因子-β(TGF-β)显著下调;损伤中心两侧1 mm内血管密度显著下降,血管内皮生长因子(VEGF)表达水平显著下调,空洞面积显著增加,空洞周围存活神经元数量显著减少。BMS评分显示PD1敲除小鼠的自发运动功能恢复显著差于WT小鼠。结论:PD1敲除加重小鼠脊髓挫伤后的继发性损伤,增强PD1信号可能有助于促进脊髓损伤后的运动功能恢复。
Objective:This study is aimed to explore the effects of programmed death protein 1(PD1)knockout on the polarization of macrophage/microglia,angiogenesis,neuron survival,cavity formation and motor function after spinal cord injury(SCI),and to clarify the roles of PD1 in the development of secondary injury,hoping to lay the foundation for bringing up intervention measures in the future.Methods:Spinal cord contusion was made in PD1 knockout mice.Immunofluorescence staining was used to study the polarization,angiogenesis,neuronal survival and cavity formation.Real time RT-PCR and Western Blot were used to evaluate the expression of inflammatory factors and vascular endothelial growth factor(VEGF).Basso Mouse Scale(BMS)score was adopted to assess the motor function of the animals.Results:Compared with that in wild-type(WT)mice,the expression of M1 macrophage/microglia associated factors such as iNOS,CD86,IL-6,and TNF-αin the injured spinal cord of PD1 deficient mice were significantly increased.The levels of M2 macrophage/microglia associated molecules,such as Arg1,IL-10,IL-4 and TGF-β,were significantly down-regulated.The vascular density within and around the lesion area of PD1 deficient spinal cord decreased significantly.Accordingly,the expression of VEGF was decreased,the cavity area was enlarged,and the number of surviving neurons around the cavity was reduced significantly.In addition,BMS scoring revealed that the recovery of spontaneous motor function of PD1 knockout mice was significantly worse than that of WT mice.Conclusion:PD1 knockout worsens the secondary spinal cord injury.Amplifying PD1 signaling may be beneficial for promoting locomotion recovery following SCI.
作者
唐润妍
惠首智
张明
张倩
吴寅
张坤
Tang Runyan;Hui Shouzhi;Zhang Ming;Zhang Qian;Wu Yin;Zhang Kun(Class 2018,the eighth team of the Second Battalion,School of Basic Medicine,Air Force Medical University,Xi’an 710032;Class 2018,the fifth team of the Second Battalion,School of Basic Medicine,Air Force Medical University,Xi’an 710032;Department of Neurobiology,School of Basic Medicine,Air Force Medical University,Xi’an 710032;Department of Neurology,Hainan Hospital of Chinese People’s Liberation Army General Hospital,Hainan 572013;Xi’an Gaoxin Hospital affiliated to Northwest University,Xi’an 710075,China)
出处
《神经解剖学杂志》
CAS
CSCD
2021年第5期517-524,共8页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金(81601056,81901252)
陕西省重点研发计划(2020SF-083)
陕西省自然科学基金基础研究计划(2020-JM-722)。
