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目的探讨锌对第5腰椎(L5)脊神经结扎横切所致神经病理性疼痛(NPP)模型的学习记忆功能障碍及海马组织中P53表达的影响。方法成年小鼠(C57/BL6)48只,随机分为3组(n=16),对照组(con),NPP组及高锌NPP组(Zn-NPP)。对照组仅暴露L5脊神经不结扎,NPP组施行L5脊神经左侧结扎横切(L L5-SNT)手术,Zn-NPP组施行左侧L5-SNT手术后饮用硫酸锌水溶液。3组术后第29天进行Morris水迷宫实验测量小鼠空间学习记忆能力,水迷宫实验结束后,采用流式细胞术检测各组细胞凋亡率,采用免疫组织化学及免疫印迹法检测各组P53表达。结果 Morris水迷宫实验中,NPP组(21.03±12.08)m/s,Zn-NPP组(16.97±8.59)m/s及con组(8.74±3.61)m/s,隐藏平台潜伏期逐渐缩短(P<0.05),并穿过平台所在象限的时间及路径占总游泳时间和总路径的比例逐渐增加(P<0.05)。流式细胞术检测结果显示,con组(1.88%),Zn-NPP(16.12%)及NPP组(33.17%)凋亡率逐渐增多(P<0.05)。免疫组织化学结果显示,con组,Zn-NPP组及NPP组的P53阳性表达吸光度值在CA1区分别为27.42±7.08、37.52±10.58及53.31±9.26;CA3区分别为29.51±8.31、36.21±7.91及56.71±8.09。免疫印迹结果显示,con组,Zn-NPP组及NPP组的P53表达吸光度值分别为0.12±1.01、0.37±1.13和0.86±1.22,吸光度值逐渐增多(P<0.05)。结论锌可改善NPP后小鼠学习记忆能力。
Objective To investigate the effects of zinc on learning and memory dysfunction and P53 expression in hippocampal tissue of neuropathic pain (NPP) model induced by lumbar spinal cord transection in the 5th lumbar vertebra (L5). Methods Forty-eight adult C57 / BL6 mice were randomly divided into three groups (n = 16), control group, NPP group and Zn-NPP group. In the control group, only the L5 spinal nerve was not ligated, the L5-SNT L5 spinal nerve was performed in the NPP group, and the L-5 SNT was used in the Zn-NPP group. The Morris water maze test was used to measure the spatial learning and memory abilities of the mice in the three groups at the 29th day after operation. After the water maze test, the apoptosis rate of each group was detected by flow cytometry. The immunohistochemistry and immunoblotting were used to detect the groups P53 expression. Results In the Morris water maze test, the latent period of hidden platform was shortened gradually in the NPP group (21.03 ± 12.08) m / s, the Zn-NPP group (16.97 ± 8.59) m / s and the con group (8.74 ± 3.61) m / 0.05). The proportion of time and path traversing the quadrant in which the platform is located to total swimming time and total path gradually increased (P <0.05). The results of flow cytometry showed that the apoptotic rates of con group (1.88%), Zn-NPP (16.12%) and NPP group (33.17%) increased gradually (P <0.05). The results of immunohistochemistry showed that the positive expression of P53 in con group, Zn-NPP group and NPP group were 27.42 ± 7.08,37.52 ± 10.58 and 53.31 ± 9.26 in CA1 and 29.51 ± 8.31,36.21 ± 7.91 in CA3 And 56.71 ± 8.09. Immunoblotting results showed that the absorbance values of P53 in con group, Zn-NPP group and NPP group were 0.12 ± 1.01,0.37 ± 1.13 and 0.86 ± 1.22 respectively, and the absorbance values gradually increased (P <0.05). Conclusion Zinc can improve learning and memory ability of mice after NPP.