摘要
目的观察α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)谷氨酸受体-2(AMPA-GluR2)在大鼠脊髓损伤(SCI)后少突胶质前体细胞(OPCs)凋亡中的作用。方法原代培养OPCs,与脊髓组织块进行共培养。应用Western blot检测OPCs中AMPAR-GluR2表达,细胞流式技术检测OPCs凋亡。SD大鼠随机分为假手术组、SCI组,NBQX拮抗剂组和JSTx拮抗剂组(n=15)。Western blot检测脊髓组织中AMPA-GluR2的表达,原位缺口末端标记法(TUNEL)法检测OPCs的凋亡,动物后肢神经功能评分[Basso-Beattie-Bresnahan(BBB)评分]评估运动功能恢复。结果与SCI组织块共培养的OPCs凋亡率(36.5%)高于正常对照组(6.3%)(P<0.01),且AMPAR-GluR2在24 h和48 h表达SCI组(0.72±0.02,0.51±0.03)明显低于对照组(0.90±0.04,0.91±0.02)(P<0.01)。动物实验结果显示:OPCs凋亡指数:假手术组(0.13±0.01)<JSTx组(0.17±0.01)<NBQX组(0.21±0.02)<SCI组(0.42±0.02)(P<0.05);AMPA-GluR2表达:假手术组(0.94±0.07)>SCI组(0.69±0.03)>JSTx组(0.54±0.12)>NBQX组(0.37±0.07)(P<0.05);SCI后7 d BBB评分:假手术组(21.00±0.00)分>NBQX组(8.90±0.82)分和JSTx组(9.50±0.79)分>SCI组(6.50±0.61)分(P<0.05),NBQX组和JSTx组之间差异无统计学意义(P>0.05)。结论SCI后OPCs凋亡与AMPAR-GluR2表达减少有关。
Objective To investigate the effect of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor (AMPA)-glutamate receptor subunit-2 (AMPA-GluR2) on the apoptosis of oligodendrocyte precursor cells (OPCs) after spinal cord injury (SCI) in rats. Methods The primary OPCs were co cultured with spinal cord tissue after SCI model was establishedusing Allen’s method. The apoptosis was detected by flow cytometry, and the AMPAR-GluR2 expression was detected by Westem-blot.60 female SD rats were randomly divided into sham operation group, SCI group, 6-nitro-2, 3-dioxo-1, 2, 3, 4-tetrahydrobenzo [f]quinoxaline-7-sulfonamide (NBQX) group and JSTx group (n=15). Western blotting was used to detect AMPA-GluR2 expression and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) method was used to detect OPCs apoptosis in spinal cord. Basso-Beattie-Bresnahan test (BBB score) was used to evaluate the recovery of motor function. Results The co cultured OPCs apoptosis rate of SCI group (36.5%) was higher than the control group (6.3%)(P< 0.01), and GluR2 in 24 h and 48 h expression in SCI group (0.72±0.02, 0.51±0.03)was less than that of control group (0.90±0.04, 0.91±0.02)(P<0.01);The apoptosis index of OPCs in spinal cord: Sham operation group (0.13±0.01)<JSTx group (0.17±0.01)<NBQX group (0.21±0.02)<SCI group (0.42±0.02)(P<0.05). AMPA-GluR2 expression: Sham operation group (0.94±0.07)>SCI group (0.69±0.03)>JSTx group (0.54±0.12)>NBQX group (0.37±0.07)(P<0.05). The BBB score: Sham operation group (21.00±0.00)>JSTx group (9.50±0.79)and NBQX group (8.90±0.82)>SCI group (6.50±0.61)(P<0.05). And there was no difference between NBQX groupand JSTx group (P>0.05). Conclusion The apoptosis of OPCs after spinal cord injury in rats is related to the decrease of AMPA-GluR2 expression.
作者
周强
李超
王学辉
Zhou Qiang;Li Chao;Wang Xuehui(Department of Orthopedics, Tianjin First Center Hospital, Tianjin 300192, China;Department of Cervical and Lumbar Pain, Weifang Hospital of Traditional Chinese Medicine, Weifang 261041, China;Graduate School of Tianjin Medical University, Tianjin 300070, China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第4期722-725,共4页
Chinese Journal of Experimental Surgery
基金
天津市卫计委科技基金(2015KZ036).
