丙戊酸钠减轻大鼠神经病理性疼痛及其脊髓水平机制
Sodium valproate relieves neuropathic pain and its mechanism at the spinal cord level in rats
左慧颖 1邵翠杰 2束宁德 3熊桃 4范倩倩 4李青青 4王德强5
作者信息
- 1. 滨州医学院烟台附属医院康复医学科,烟台市 264100
- 2. 滨州医学院附属医院医学研究中心,滨州市 256600
- 3. 滨州医学院附属医院神经内科,滨州市 256600
- 4. 滨州医学院附属医院康复科,滨州市 256600
- 5. 滨州医学院附属医院疼痛科,滨州市 256600
- 折叠
摘要
目的 探讨腹腔注射组蛋白去乙酰化酶(HDACs)抑制剂丙戊酸钠(VPA)对神经病理性疼痛(NP)的镇痛作用及其脊髓水平镇痛机制.方法 实验1:将30只SD雄性大鼠按照随机数字表法以1:4比例将其分为假手术组(Sham组,n=6)、慢性压缩性损伤组(CCI组,n=24).CCI组行坐骨神经压迫术,Sham组仅分离坐骨神经周围组织.根据预实验中行为学观察的结果对CCI组大鼠再一次随机分组,作为CCI组的4个亚组:5、10、15和20日组.实验2:将24只SD雄性大鼠按照随机数字表法随机分为为假手术+生理盐水治疗组(Sham+NS组)、假手术+丙戊酸钠治疗组(Sham+VPA组)、慢性压缩性损伤+生理盐水治疗组(CCI+NS组)、慢性压缩性损伤+丙戊酸钠治疗组(CCI+VPA),各组6只.实验1、2各组均于术前1 d,术后24h后每隔1日测定机械缩足反射阈值(MWT)及热缩足反射潜伏期(TWL).其中实验2各组大鼠于术后第3日开始通过腹腔注射给药,1次/d,连续7d.术后不同时点断头处死并取大鼠脊髓腰膨大处(L4～L6)组织,分别采取PCR方法测定HDACs以及GABA的mRNA水平,蛋白质印迹法测定相应的蛋白水平.结果 与Sham组相比,CCI组MWT、TWL均明显降低(P均＜0.05).CCI组HDAC2、3的mRNA和蛋白表达均高于Sham组(P均＜0.05),AcH3的蛋白表达,GABA的mRNA和蛋白表达均低于Sham组(P均＜0.05).经过治疗后,CCI+VPA组较CCI+NS组,HDAC1、2、3的mRNA含量均降低(P均＜0.05),HDAC2蛋白表达也降低(P＜0.05),但AcH3、GABA的蛋白表达均增加(P均＜0.05).结论 腹腔注射VPA可在脊髓水平降低NP大鼠HDACs的mRNA和蛋白表达,而达到缓解NP的作用,其可能的作用机制与促进升高AcH3、GABA蛋白的表达有关.
Abstract
Objective To investigate the analgesic effect of intraperitoneal injection of histone deacetylase(HDACs)inhibitor valproate acid sodium(VPA)on neuropathic pain(NP)and its analgesic mechanism at spinal level.Methods In experiment 1,30 male SD rats were randomly divided into sham-operation group(Sham group,n=6)and chronic compression injury group(CCI group,n=24)at a ratio of 1:4.The sciatic nerve compression was performed in the CCI group,while only surrounding tissue of the sciatic nerve was separated in the Sham group.Twenty-four rats were again randomly assigned to four subgroups of the CCI group:the 5-day group,the 10-day group,the 15-day group,and the 20-day group based on behavioral observations in the preliminary experiment.In experiment 2,24 male SD rats were randomly divided into Sham+NS group,Sham+VPA group,CCI+NS group and CCI+VPA group(n=6)according to random number method.The mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were measured respectively on the first day before operation and 24 hours after operation in all groups.In experiment 2,the drugs were injected intraperitoneally 3 days after operation,once a day for 7 consecutive days.The rats were sacrificed by amputation at different time point after operation,and the tissues from the lumbar enlargement of the spinal cord(L4-L6)were collected.The mRNA levels of HDACs and GABA were determined by PCR,and the corresponding protein levels were detected by western blotting.Results Compared with the Sham group,MWT and TWL decreased significantly in the CCI group and CCI+NS group(all P＜0.05).At the spinal cord level,the mRNA and protein expressions of HDAC2 and HDAC3 in the CCI group were higher than those in the Sham group(all P＜0.05).The protein expression of AcH3,the mRNA and protein expression of GABA were lower than those in the Sham group(all P＜0.05).The mRNA content of HDAC1,2 and 3 in the CCI+VPA group was lower than that in the CCI+NS group(all P＜0.05).The protein expression of HDAC 2 decreased(P＜0.05),but the protein expression of AcH3 and GABA increased(all P＜0.05).Conclusion Intraperitoneal injection of HDAC inhibitors VPA can reduce the mRNA and protein expression of HDACs in NP rats at the spinal cord level,and alleviate the effect of NP,which may be related to the increase of AcH3 and inhibitory neurotransmitter GABA protein expression.
关键词
神经痛/丙戊酸钠/组蛋白乙酰化/γ-氨基丁酸Key words
Neuralgia/Valproic acid/Histone acetylation/Gamma-aminobutyric acid引用本文复制引用
基金项目
山东省自然科学基金(ZR2018LH005)
出版年
2024
NETL
NSTL
万方数据