鞘内注射白藜芦醇对骨癌痛大鼠脊髓背角神经元CaMKⅡ活化的影响  

Effect of intrathecal resveratrol on activation of CaMK Ⅱ in spinal dorsal horn neurons of rats with bone cancer pain

作　　者：张志辉[1] 徐俊[1] 柯建辉[1] 马淑娟[1] 胡学铭[2] 

机构地区：[1]江苏省丹阳市第二人民医院麻醉科,212300 [2]徐州医学院附属医院疼痛科

出　　处：《中华麻醉学杂志》2015年第1期68-71,共4页Chinese Journal of Anesthesiology

摘　　要：目的 评价鞘内注射白藜芦醇对骨癌痛大鼠脊髓背角神经元钙-钙调素依赖性蛋白激酶Ⅱ（CaMKⅡ）活化的影响.方法 清洁级成年雌性SD大鼠32只,体重180～220 g,采用随机数字表法分为4组（n=8）：假手术组（S组）、骨癌痛组（B组）、骨癌痛＋白藜芦醇组（BR组）和骨癌痛＋溶媒对照组（BD组）.B组、BR组和BD组右侧胫骨骨髓腔注射Walker 256乳腺癌细胞5μl（4＆#215;105个/ml）;S组右侧胫骨骨髓腔注入5μl生理盐水.于造模后第12、13和14天,BR组鞘内注射白藜芦醇200 μg/10μl,1次/d;BD组鞘内注射1％二甲基亚砜10μl,1次/d.于注入乳腺癌细胞前（To）、注入乳腺癌细胞后3、5、7、10、12和14 d（T1-6）时测定机械痛阈和热痛阈,并于T6时取右侧胫骨,用HE染色法检测胫骨病理学变化;取大鼠L4,5脊髓节段,用免疫荧光法测定脊髓背角神经元磷酸化CaMKⅡ（p-CaMKⅡ）的细胞定位,并用免疫印迹法检测其蛋白表达.结果 与S组比较,B组、BR组和BD组T2-6时机械痛阈和热痛阈降低,T6时脊髓p-CaMKⅡ表达上调（P＜0.01）,且主要与神经元共表达;与B组比较,BR组T5,6时机械痛阈和热痛阈升高,T6时脊髓p-CaMKⅡ表达下调（P＜0.01）,BD组各时点机械痛阈和热痛阈以及p-CaMKⅡ表达差异无统计学意义（P＞0.05）.结论 鞘内注射白藜芦醇减轻骨癌痛大鼠痛觉过敏的机制可能与抑制脊髓背角神经元CaMKⅡ活化有关.Objective To evaluate the effect of intrathecal resveratrol on activation of Ca2＋/ calmodulin-dependent protein kinase Ⅱ （CaMK Ⅱ） in spinal dorsal horn neurons of rats with bone cancer pain.Methods Thirty-two adult female Sprague-Dawley rats,weighing 180-220 g,were equally randomized into 4 groups using a random number table：sham operation group （group S）,bone cancer pain group （B group）,and bone cancer pain ＋ solvent control group （BD group）.Walker 256 mammary gland cancer cell suspension （4＆#215; 105 cells/ml） 5 μl was injected into the medullary cavity of the right tibia in B,BR and BD groups.Normal saline 5 μl was given in group S.On 12,13 and 14 days after injection of mammary gland cancer cells,resveratrol 200 μg/10 μl was injected intrathecally once a day in group BR,and 1% dimethyl sulfoxide 10 μl was intrathecally injected once a day in group BD.Before injection of mammary gland cancer cells （T0） and on 3,5,7,10,12 and 14 days after injection of mammary gland cancer cells （T1-6）,mechanical paw withdrawal threshold （MWT） and thermal paw withdrawal latency （TWL） were measured.The rats were then sacrificed and L4,5 segments of the spinal cord were removed for confirmation of the location of phosphorylated CaMK Ⅱ （p-CaMK Ⅱ） in spinal dorsal horn neurons （by immunofluorescence） and for detection of p-CaMK lⅡ expression （using Western blot）.Results Compared with group S,MWT and TWL were significantly decreased at T2-6,and p-CaMK Ⅱ expression was upregulated at T6,and p-CaMK Ⅱ was mainly co-expressed with neurons in B,BR and BD groups.Compared with group B,MWT and TWL were significantly increased at T5,6,and p-CaMK Ⅱ expression was down-regulated at T6 in group BR.There was no significant difference in MWT,TWL,and p-CaMK Ⅱ expression at each time point between group B and group BD.Conclusion Resveratrol can alleviate hyperalgesia in rats with bone cancer pain and inhibited activation of CaMK Ⅱ in spinal dorsal horn neurons may be i

关 键 词：酚类 注射 鞘内 骨肿瘤 疼痛 脊髓 神经元 钙-钙调素依赖性蛋白激酶2型 

分 类 号：R738.1[医药卫生—肿瘤] R614[医药卫生—临床医学]