AMPK-依赖性自噬信号通路在氯胺酮减轻大鼠糖尿病神经痛中的作用  

Role of AMPK-dependent autophagic signaling pathway in ketamine-induced reduction of diabetic neuropathic pain in rats

作　　者：丁洁[1] 胡益民[4] 杨春[1] 朱滨[2] 华飞[3] 

机构地区：[1]苏州大学附属第三医院麻醉科,常州市213003 [2]苏州大学附属第三医院重症医学科,常州市213003 [3]苏州大学附属第三医院内分泌代谢科,常州市213003 [4]南京军区南京总医院麻醉科,210002

出　　处：《中华麻醉学杂志》2016年第2期180-182,共3页Chinese Journal of Anesthesiology

摘　　要：目的评价腺苷酸活化蛋白激酶（AMPK）-依赖性自噬信号通路在氯胺酮减轻大鼠糖尿病神经痛中的作用。方法清洁级健康雄性Wistar大鼠60只，3月龄，体重200～250g，采用随机数字表法分为5组（n=12）：对照组（C组）、生理盐水组（NS组）、氯胺酮组（K组）、氯胺酮＋Com—poundC组（KC组）和氯胺酮＋3-甲基腺嘌呤（3-MA）组（KM组）。NS组、K组、KC组和KM组单次腹腔注射链脲菌素65mg／kg制备大鼠糖尿病神经痛模型。4周后，NS组、K组、KC组和KM组分别腹腔注射等容量生理盐水、氯胺酮10mg／kg、氯胺酮10mg，kg＋CompoundC1mg／kg和氯胺酮＋3-MA2μl，1次／d，连续7d。药物干预后第8天，测定机械缩足反应阈（MWT），随后处死取L。脊髓，采用Western blot法检测背根神经节AMPKa、Beclin-1、自噬微管相关蛋白轻链（LC）3B的表达，并测定树突棘密度。结果与C组比较，NS组MWT、脊髓背根神经节AMPKα、Beclin-1、LC3B表达水平和树突棘密度降低（P〈0．05）；与NS组比较，K组MWT、脊髓背根神经节AMPKα、Beclin-1、LC3B表达水平和树突棘密度升高（P〈0．05）；与K组比较，KC组和KM组MWT、脊髓背根神经节AMPKα、Beclin-1、LC3B表达水平和树突棘密度降低（P〈0．05）。结论AMPK-依赖性自噬信号通路的激活参与了氯胺酮减轻大鼠糖尿病神经痛的过程。Objective To evaluate the role of AMP-activated protein kinase （AMPK） -dependent autophagic signaling pathway in ketamine-induced reduction of diabetic neuropathic pain （DNP） in rats. Methods Sixty male Wistar rats, aged 3 months, weighing 200-250 g, were equally randomized into 5 groups using a random number table： control group （ C group） , normal saline group （ NS group） , ketamine group （K group） , ketamine ＋ Compound C group （KC group） , and ketamine ＋ 3-methyladenine （3-MA） group （KM group）. DNP model was established by intraperitoneal injection of streptozocin （STZ） 65 mg/kg in anesthetized rats. Four weeks later, the equal volume of normal saline, ketamine 10 mg/kg, ketamine 10 mg/kg ＋ Compound C 1 mg/kg, and ketamine ＋ 3-MA 2 μl were injected intraperitoneally once a day for 7 consecutive days in NS, K, KC and KM groups, respectively. The mechanical paw with- drawal threshold （MWT） was measured on 8th day. The rats were then sacrificed, and the lumbar segment （L1-5） of the spinal cord was removed for determination of the expression of AMPKα, Beclin-1, microtubule-associated protein 1 light chain （LC） 3B （by Western blot） and dendritic spine density in the dorsal root ganglia. Results Compared with group C, the MWT, expression of AMPKα, Beclin-1, and LC3B,and dendritic spine density were significantly decreased in group NS （P〈0.05）. Compared with group NS, the MWT, expression of AMPKα, Beclin-1, and LC3B, and dendritic spine density were significantly increased in group K （P〈0. 05）. Compared with group K, the MWT, expression of AMPKα, Beclin-1, and LC3B, and dendritic spine density were significantly decreased in KC and KM groups （P〈0.05）. Conclusion Activation of AMPK-dependent autophagic pathway is involved in ketamine-induced reduction of DNP in rats.

关 键 词：蛋白质丝氨酸苏氨酸激酶 自噬 氯胺酮 糖尿病 神经痛 

分 类 号：R587.2[医药卫生—内分泌] R741[医药卫生—内科学]