紫杉醇诱发痛大鼠脊髓背角超极化活化环核苷酸门控通道4的表达及其与γ-氨基丁酸B型受体的关系  

Expression of hyperpolarization-activated cyclic nucleotide-gated 4 channel in the spinal dorsal horn of rats with paclitaxel-induced neuropathic pain and its relationship withγ-aminobutyric acid B receptor

作　　者：田晓琳 郭跃先[2] 刘朋[1] 李昭[1] 刘欣[1] 赵爽[1] 王秀丽[1] 王贵英 Tian Xiaolin;Guo Yuexian;Liu Peng;Li Zhao;Liu Xin;Zhao Shuang;Wang Xiuli;Wang Guiying(Department of Anesthesiology,the Third Affiliated Hospital of Hebei Medical University,Shijiazhuang 050051,China;Department of Urology,the Third Affiliated Hospital of Hebei Medical University,Shijiazhuang 050051,China;Department of Gastroenterological Surgery,the Third Affiliated Hospital of Hebei Medical University,Shijiazhuang 050051,China)

机构地区：[1]河北医科大学第三医院麻醉科,石家庄050051 [2]河北医科大学第三医院泌尿外科,石家庄050051 [3]河北医科大学第三医院胃肠外科,石家庄050051

出　　处：《中华实验外科杂志》2020年第4期708-712,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81371231、81971001);河北省自然科学基金(H2018206260)。

摘　　要：目的探讨紫杉醇诱发神经痛大鼠脊髓背角超极化活化环核苷酸门控通道4(HCN4)的表达变化及其与γ-氨基丁酸B型(GABAB)受体的关系。方法清洁级SD雄性大鼠(购自河北医科大学动物实验中心),69周龄,体重180~220 g。于实验开始第1、3、5、7 d分别腹腔注射紫杉醇2 mg/kg制备神经痛模型,行鞘内置管术成功后,随机数字表法将其分为5组(n=10):紫杉醇模型组(P组),紫杉醇模型+生理盐水组(N组),紫杉醇模型+慢病毒空载体组(BV组),紫杉醇模型+HCN4通道慢病毒组(H组),紫杉醇模型+ZD7288组(I组),10只同龄大鼠作为空白对照组(C组)。腹腔注射紫杉醇后15 d鞘内注射药物20 μl:N组注射生理盐水,BV组注射慢病毒空载体(1×108 TU/ml),H组注射HCN4通道慢病毒(1×108 TU/ml)。腹腔注射紫杉醇后21 d鞘内注射药物20 μl:I组注射HCN4通道抑制剂ZD7288(30 μg)。腹腔注射紫杉醇前1 d(T0)、注射后第14天(T1)、第21天(T2)时点分别测定50%机械缩足反应阈(MWT)。于T2时点取脊髓背角组织标本,采用免疫组织化学法和蛋白质印迹法(Western blot)法测定HCN4通道和GABAB受体的蛋白表达水平。测量数据采用均值±标准差(Mean±SD)表示,组内比较采用重复测量设计的方差分析,组间比较采用单因素方差分析。结果与T0时点[(15.4±2.7)、(14.8±2.1)、(14.6±2.1)、(14.2±2.2)、(15.2±2.1) g]比较,P组、N组、BV组、H组、I组T1时[(4.8±1.3)、(4.7±1.5)、(4.8±1.1)、(5.0±1.3)、(4.8±1.3) g] MWT均降低(t=11.326、12.177、12.932、11.298、13.449,P<0.05),差异有统计学意义;与T1时点比较,H组、I组T2时点[(9.9±1.0)、(10.3±1.5) g]MWT均升高(t=9.389、9.006,P<0.05),差异有统计学意义。与C组[(14.4±2.6) g]比较,T1时点P组、N组、BV组、H组、I组MWT均降低(t=10.394、10.009、10.537、10.117、10.335,P<0.05),差异有统计学意义;与P组[(5.2±1.7) g]比较,T2时点H组和I组[(9.9±1.0)、(10.3±1.5) g]MWT均升高(t=7.440、7.212,P<0.05),差异�Objective To investigate the expression of hyperpolarization-activated cyclic nucleotide-gated 4(HCN4)channel in the spinal dorsal horns of rats with paclitaxel-induced neuropathic pain and its relationship withγ-aminobutyric acid B(GABAB)receptor.Methods The male SD rats(Purchased from animal experiment center of Hebei Medical University)of clean grade aged 69 weeks,weighing 180-220 g.The model of paclitaxel-induced neuropathic pain was established by intraperitoneal injection of 2 mg/kg of paclitaxel 1,3,5 and 7 days after the experiment.After successful intrathecal catheterization,the rats were randomly divided into 5 groups(n=10):Paclitaxel model group(P group),paclitaxel model+saline(N),paclitaxel model+lentivirus empty vector group(BV group),paclitaxel model+HCN4 channel lentivirus group(H group),paclitaxel model+ZD7288 group(I group),10 rats of the same age as the blank control group(C group).On the 15th day after intraperitoneal injection,different drugs were intrathecally injected with 20μl:Group N was injected with normal saline,group BV was injected with HCN4 channel lentivirus empty vector(1×108 TU/ml),and group H was injected with HCN4 channel lentivirus(1×108 TU/ml);On the 21th day after intraperitoneal injection:group I was injected with HCN channel inhibitor ZD7288(30 g/20 L).The 50%mechanical foot shrinkage threshold(MWT)was measured 1 day before intraperitoneal injection(T0),14 days(T1)and 21 days(T2)after injection.The expression of HCN4 channel and GABAB receptor in the spinal dorsal horn was determined by immunohistochemistry and Western blotting 21 days(T2)after intraperitoneal injection.The measured data were expressed as meanstandard deviation.Statistical analyses were performed using the One-Way ANOVA in inter-group comparison,the Repeated Measures ANOVA in intra-group comparison.The male SD rats(Purchased from animal experiment center of Hebei Medical University)of clean grade aged 69 weeks,weighing 180-220 g.The model of paclitaxel-induced neuropathic pain was established by intrape

关 键 词：紫杉醇 神经痛 超极化激活环核苷酸门控通道4 慢病毒感染 γ-氨基丁酸B型受体 

分 类 号：R741[医药卫生—神经病学与精神病学]