目的:探讨Akt3基因敲除对坐骨神经结扎小鼠神经病理性痛影响的机制。方法:SPF级C57BL/6小鼠为背景的Akt3基因敲除小鼠(Akt3n -/-小鼠)12只按照随机数字表法分为Akt3基因敲除＋假手术组(Akt3n -/-＋Sham组，n n＝6)，Akt3基因敲除＋模型组(Akt3n -/-＋CCI组，n n＝6)；野生型小鼠(WT小鼠)12只采用随机数字表法分为野生＋Sham组(WT＋假手术组，n n＝6)，野生＋模型组(WT＋CCI组，n n＝6)。CCI组采用坐骨神经慢性结扎损伤制作神经病理性疼痛模型，Sham组行假手术。各组小鼠分别于术前1 d，CCI术后7 d测定机械刺激缩足反应阈值(paw withdrawal mechanical threshold，PWMT)。Sham组于造模后7 d，CCI组于造模后7 d、14 d后各处死3只小鼠，取L3～5脊髓节段，采用Western blot方法检测Akt3、磷酸化Akt(p-Akt)、GSK3β、磷酸化NR2B(p-NR2B)表达水平。采用SPSS 21.0软件进行统计学分析。n 结果:小鼠PWMT，脊髓组织Akt3、p-Akt、GSK3β、p-NR2B蛋白表达的组别×时间交互效应均显著(n F＝16.667, 269.899，26.651，572.998，37.836，n P<0.01)。与Akt3n -/-＋Sham组比较，Akt3n -/-＋CCI组术后7 d PWMT降低[(0.34±0.20)g，(1.18±0.11)g；n P<0.01]，脊髓水平p-Akt[ (0.90±0.08)，(0.51±0.06)；n P<0.01]、GSK3β[ (0.74±0.04)，(0.29±0.02)；n P<0.01]、p-NR2B[(0.96±0.11)，(0.71±0.04)；n P<0.05]表达增加。与WT＋CCI组比较，Akt3n -/-＋CCI组术后7 d PWMT降低[(0.34±0.20)g，(0.49±0.12)g；n P<0.05]；脊髓水平p-Akt表达减少[ (0.90±0.08)，(1.02±0.17)；n P<0.05]、GSK3β[ (0.74±0.04)，(0.57±0.09)；n P<0.01]、p-NR2B表达增加[ (0.96±0.11)，(0.91±0.08)；n P<0.05]，差异有统计学意义。n 结论:Akt3基因敲除后，小鼠坐骨神经结扎神经病理性疼痛加重可能与Akt/GSK3β/NR2B表达变化有关。“,”Objective:To explore the mechanism of Akt3 gene knockout on neuropathic pain induced by sciatic nerve ligation.Methods:Twelve Akt3 knockout mice with SPF grade C57BL/6 mice as background were randomly divided into Akt3 gene knockout + Sham group (Akt3n -/-+ Sham, n n=6)and Akt3 gene knockout + CCI group(Akt3n -/-+ CCI, n n=6). Twelve wild type C57BL/6 mice were randomly divided into wild + Sham group (WT + Sham, n n=6) and wild + CCI group (WT + CCI, n n=6) by random number table. Chronic sciatic nerve ligation was used to make neuropathic pain model in CCI group, and sham group was subjected to sham operation. The paw withdrawal mechanical threshold (PWMT) was measured 1 day before operation and 7 days after CCI. Three mice were randomly killed in Sham group 7 days after modeling, and 3 mice in CCI group were killed at 7 days and 14 days respectively.The L3-5 spinal cord segment was taken. Akt3, phosphorylated Akt (p-Akt), GSK3β and phosphorylated NR2B (p-NR2B) were detected by Western blot. SPSS 21.0 software was used for statistical analysis.n Results:Group time interaction effects of PWMT, the expression of protein Akt3, p-Akt, GSK3β, p-NR2B in spinal cord were significantly different (n F=16.667, 269.899, 26.651, 572.998, 37.836, n P<0.01). Compared with the Akt3n -/-+ Sham group, the PWMT in Akt3n -/-+ CCI group was significantly lower((0.34±0.20)g, (1.18±0.11)g, n P<0.01), and the expression of p-Akt((0.90±0.08), (0.51±0.06),n P<0.01), GSK3β((0.74±0.04), (0.29±0.02),n P<0.01) and p-NR2B((0.96±0.11), (0.71±0.04),n P<0.05) in spinal cord increased. Compared with the WT+ CCI group, the PWMT of the Akt3n -/-+ CCI group was obviously lower((0.34±0.20)g , (0.49±0.12)g, n P<0.05), and the expression of p-Akt((0.90±0.08), (1.02±0.17),n P<0.05)decreased, and the expression of GSK3β((0.74±0.04), (0.57±0.09),n P<0.01) and p-NR2B((0.96±0.11), (0.91±0.08),n P<0.05) increased.n Conclusion:After Akt3 gene knockout, the aggravation of neuropathic pain after sciatic nerve ligation may be related with the change of Akt/GSK3β/NR2B expression.