Liu Qing, Xu Xiaohan, Xu Li, Huang Yuguang
Department of Anesthesiology,Plastic Surgery Hospital,CAMS and PUMC,Beijing 100144,China.
Department of Anesthesiology,PUMC Hospital,CAMS and PUMC,Beijing 100730,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2020 Jun 30;42(3):338-346. doi: 10.3881/j.issn.1000-503X.12002.
To observe the cell origin of N-methyl-D-aspartic acid(NMDA)receptor expression in skin after chronic ischemic pain modeling in rats and explore the role of NMDA receptor in type Ⅰ complex regional pain syndrome. Forty-two adult male Sprague-Dawley rats were randomly divided into five groups:sham operation group(=12),chronic post ischemia pain(CPIP)group(=12),CPIP+normal saline(NS)group(=6),CPIP+NMDA group(=6),and CPIP+MK801 group(=6).Six rats in the sham operation group and CPIP group were sacrificed under deep anesthesia one day after modeling.The plantar skin and L3-L5 spinal cord tissue were used for NR1(NMDA receptor)subunit immunofluorescence detection and for Western blotting of NR1,interleukin(IL)-1β,and tumor necrosis factor(TNF)-α.For the remaining rats,the mechanical withdrawal threshold(MWT)values on the 2nd,6th,10th and 14th day after ischemia were recorded,and the corresponding drugs were injected subcutaneously from the 6th day after ischemia.The skin and L3-L5 spinal cords were collected on the 14th day,and the same detection methods were applied. Compared with the sham operation group,the CPIP group had significantly higher expressions of NR1(1.708±0.064;=12.120, <0.001),IL-1β(2.575±0.305;=5.158, =0.003),and TNF-α(2.691±0.217;=7.786, <0.001)in the skin on the first day after modeling.After intervention with NMDA and MK801,the MWT value was [(20.37±0.95)g] in the CPIP+NS group,which was significantly higher than that in CPIP+NMDA group [(15.85±1.09)g;=10.920, <0.001] but significantly lower than that in CPIP+MK801 group[(22.95±0.96)g;=6.421, <0.001] 10 days after modeling.On the 14th day,compared with the MWT of the CPIP+NS group [(21.57±0.96)g],the CPIP+NMDA group had significantly decreased MWT value [(16.53±1.63)g;=12.190, <0.001],and the CPIP+MK801 group had significantly increased MWT value [(23.27±1.28)g;=4.094, =0.025].Compared with the sham operation group,the CPIP group had significantly increased NR1 expression(1.708±0.064;=10.910, <0.001)and the CPIP+NS group had significantly increased expressions of IL-1β(2.518±0.147;=11.010, <0.001)and TNF-α(1.949±0.184;=10.870, <0.001).Compared with the CPIP+NS group,the CPIP+NMDA group had significantly increased expressions of IL-1β(4.816±0.607;=16.670, =0.003)and TNF-α(2.629±0.349;=7.790, <0.001)and the CPIP+MK801 group had significantly decreased expressions of IL-1β(1.048±0.257;=10.660, =0.003)and TNF-α(0.790±0.165;=13.280, <0.001). NMDA receptor activation in skin keratinocytes after chronic ischemia in rats hinders the expression of inflammatory cytokines such as IL-1β and TNF-α,which may be involved in central sensitization and pain conduction of type Ⅰ complex regional pain syndrome.
观察大鼠慢性缺血性疼痛模型建立后皮肤中N-甲基-D-天冬氨酸（NMDA）受体表达的细胞来源，并探讨NMDA受体在Ⅰ型复杂性区域疼痛综合征中的作用。42只成年雄性Sprague-Dawley大鼠随机分为五组：假手术组（n = 12）、慢性缺血后疼痛（CPIP）组（n = 12）、CPIP + 生理盐水（NS）组（n = 6）、CPIP + NMDA组（n = 6）和CPIP + MK801组（n = 6）。建模后1天，对假手术组和CPIP组中的6只大鼠进行深度麻醉后处死。取足底皮肤和L3-L5脊髓组织用于NR1（NMDA受体）亚基免疫荧光检测以及NR1、白细胞介素（IL）-1β和肿瘤坏死因子（TNF）-α的蛋白质免疫印迹分析。对于其余大鼠，记录缺血后第2、6、10和14天的机械缩足阈值（MWT）值，并从缺血后第6天开始皮下注射相应药物。在第14天收集皮肤和L3-L5脊髓，采用相同的检测方法。与假手术组相比，建模后第1天CPIP组皮肤中NR1（1.708±0.064；t = 12.120，P < 0.001）、IL-1β（2.575±0.305；t = 5.158，P = 0.003）和TNF-α（2.691±0.217；t = 7.786，P < 0.001）的表达显著升高。用NMDA和MK801干预后，建模后10天CPIP + NS组的MWT值为[（20.37±0.95）g]，显著高于CPIP + NMDA组[（15.85±1.09）g；t = 10.920，P < 0.001]，但显著低于CPIP + MK801组[（22.95±0.96）g；t = 6.421，P < 0.001]。在第14天，与CPIP + NS组的MWT[（21.57±0.96）g]相比，CPIP + NMDA组的MWT值显著降低[（16.53±1.63）g；t = 12.190，P < 0.001]，CPIP + MK801组的MWT值显著升高[（23.27±1.28）g；t = 4.094，P = 0.025]。与假手术组相比，CPIP组NR1表达显著升高（1.708±0.064；t = 10.910，P < 0.001），CPIP + NS组IL-1β（2.518±0.147；t = 11.010，P < 0.001）和TNF-α（1.949±0.184；t = 10.870，P < 0.001）表达显著升高。与CPIP + NS组相比，CPIP + NMDA组IL-1β（4.816±0.607；t = 16.670，P =
