人参皂苷Rg1对神经病理性疼痛大鼠脊髓小胶质细胞活性及其p38MAPK/NF-κB表达的影响  

Effect of ginsenoside Rgl on activity of spinal microglia and expression of p38 mitogen-activated protein kinase/nucleartranscription factor-kappa B in neuropathic pain rats

作　　者：高超 郭小瑜 翁菱琳 周王琛 沈阳 於阳 韩园 

机构地区：[1]徐州医科大学临床医学系,221004 [2]徐州医科大学麻醉学院,221004 [3]徐州医科大学附属医院麻醉科,221002

出　　处：《国际麻醉学与复苏杂志》2017年第12期1084-1089,共6页International Journal of Anesthesiology and Resuscitation

基　　金：江苏省大学生创新创业训练计划（201610313034Y）

摘　　要：目的 探讨人参皂苷Rg1（ginsenoside Rg1,Rg1）对神经病理性疼痛（neuropathic pain,NP）模型大鼠热痛阈的影响,以及脊髓背角小胶质细胞活性及p38丝裂原活化蛋白激酶（p38 mitogen-activated protein kinase,p38MAPK）/NF-κB表达变化,并探讨其可能机制.方法 鞘内置管成功的36只雄性成年SD大鼠,按随机数字表法分为6组（每组6只）：假手术＋鞘内生理盐水组（A组）、慢性坐骨神经压迫损伤（chronic constrictive injury,CCI）＋鞘内生理盐水组（B组）、CCI＋鞘内低剂量Rg1组（C组）、CCI＋鞘内中剂量Rg1组（D组）、CCI＋鞘内高剂量Rg1组（E组）、CCI＋鞘内米诺环素组（F组）.A组仅切开及缝合大鼠CCI手术造模相应区域皮肤,并与其余各组同步鞘内注射生理盐水,其余各组均行CCI手术造模.每组分别于手术前1 d和手术后连续3 d鞘内给药共4次,应用Hargreaves法测定各组左右侧手术前1天及手术后第1、3、5、7、10天热刺激缩足反射潜伏期（thermal withdrawal latency,TWL）,观察不同剂量的Rg1和米诺环素对CCI大鼠热痛觉超敏的影响,并于术后第14天处死取脊髓腰膨大,采用免疫组织化学法和Western blot法分别测定小胶质细胞活化以及p38MAPK/NF-κB通路相关分子的表达情况.结果 ①鞘内注射Rg1显著抑制了CCI导致的热痛觉超敏,并且具有剂量依赖性（P〈0.05）.②鞘内注射Rg1显著抑制了CCI导致的脊髓背角小胶质细胞离子钙接头蛋白抗原（ionized calcium binding adaptor molecule-1,IBA-1）的活化以及脊髓背角小胶质细胞标记物OX-42的表达,显著抑制了小胶质细胞的过度活化,并且模拟了小胶质细胞抑制剂米诺环素的作用（P〈0.05）.③ 鞘内注射Rg1显著抑制了CCI导致的脊髓背角磷酸化p38丝裂原活化蛋白激酶（phosphorylated p38 mitogen-activated protein kinase,p-p38MAPK）和NF-κB亚单位磷酸化p65（phosphorylated p65,p-p65）的表达（P〈0.05）.结论 Rg1对CCIObjective To evaluate the effect of analgesia of ginsenoside Rg1（Rg1） in neuropathic pain（NP） and to investigate the changes in activation of both microglial cells in dorsal horn of spinal cord and p38 mitogen-activated protein kinase（p38MAPK）/NF-κB signaling pathway within microglia. Methods Thirty six rats having been intrathecally catheterized successfully were stochastically divided into 6 groups （6 rats for each group） respectively including sham＋natural saline group （group A）, chronic constrictive injury（CCI）＋natural saline group （group B）, CCI＋low-dose Rg1 group （group C）, CCI＋middle-dose Rg1 group（group D）, CCI＋high-dose Rg1 group（group E） and CCI＋minocycline group（group F）. Among all groups except group A, the left common sciatic nerve of anesthetized rats was exposed to four silk ligatures （3-0） which were tied loosely proximal to the sciatic nerveˊs trifurcation. Different medicines were injected intrathecally （i.t.） 1 d before and 1 d, 3 d and 5 d after CCI. Behavioral assessment of anti-nociceptive effects was conducted by thermal withdrawal latency（TWL） method. The content of p38MAPK, phosphorylated p65（p-p65） （member of NF-κB superfamily） and ionized calcium binding adaptor molecule-1 （IBA-1） was detected by Western blot method. The level of OX-42, one marker of microglia was examined by immunohistochemistry method. Results ① TWL values of group C, group D and group E were significantly higher than value of group B. The effect was in a dose-dependent manner （P〈0.05）. ②Compared with group B, over-activation of microglia in group C, group D and group E were markedly suppressed, indicated by deeply inhibited expression of IBA-1 and OX-42, two markers of microglia （P〈0.05）. ③ Phosphorylation of relevant molecules in p38MAPK signaling pathway in comparison to which in group B such as p38MAPK and NF-κB was attenuated abundantly in group C, group D and group E （P〈0.05）. Conclusions Activation of p38

关 键 词：人参皂苷RGL 慢性坐骨神经压迫损伤 神经病理性疼痛 p38丝裂原活化蛋白激酶/核转录因子-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]