转录因子GATA4介导电针对神经病理性痛大鼠的镇痛作用  

Spinal transcription factor GATA4 mediated adenosine A1 receptor activation contributes to electroacupuncture analgesia in neuropathic pain rats

作　　者：金文君 张明晓[1] 王雷雷[1] 戴勤学[1,2] JIN Wen-jun;ZHANG Ming-xiao;WANG Lei-lei;DAI Qin-xue(Department of Anesthesiology,The First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,Zhejiang Province,China;Department of Anesthesiology,The Second Affiliated Hospital of Zhejiang University School of Medicine,Hangzhou 310052)

机构地区：[1]温州医科大学附属第一医院麻醉科,浙江温州325000 [2]浙江大学医学院附属第二医院麻醉科,杭州310052

出　　处：《针刺研究》2020年第11期908-913,共6页Acupuncture Research

基　　金：国家自然科学基金项目(No.81704180)。

摘　　要：目的:观察转录因子GATA4是否介导电针对神经病理性痛大鼠的镇痛作用。方法:实验分两部分。第一部分将SD大鼠按照随机数字表法随机分成对照组、GATA4腺病毒干扰RNA(AV-shGATA4 RNA)组和腺病毒空载体(AV-shCTRL)组,每组6只。观察大鼠脊髓GATA4蛋白表达量,评估腺病毒AV-shGATA4 RNA的转染效率。第二部分将30只SD大鼠按照随机数字表法分成假手术组、模型组、电针组、电针复合AV-shGATA4 RNA组、电针复合AV-shCTRL组,每组6只。采用慢性坐骨神经压迫法(CCI)制备大鼠神经病理性疼痛模型。在造模后第7天电针"足三里""太冲"。在电针前48 h,将腺病毒AV-shGATA4和AV-shCTR注射到大鼠脊髓腰(L)4—L6节段。在造模前、造模后第7天和电针后60 min时检测大鼠患肢的机械痛阈值和热痛阈值;采用Western blot法检测脊髓组织中腺苷A1受体和GATA4蛋白表达量。结果:第一部分实验:对照组和AV-shCTRL组比较,大鼠脊髓GATA4蛋白表达量差异无统计学意义(P>0.05)。与AV-shCTRL组相比,AV-shGATA4 RNA组大鼠脊髓GATA4蛋白表达量明显减少(P<0.05)。各组大鼠右后肢机械痛阈值与热敏痛阈值比较差异均无统计学意义(P>0.05)。第二部分实验:在造模后第7天,除假手术组外,各组大鼠的机械痛阈值与热敏痛阈值较造模前明显下降(P<0.05)。在电针后60 min时,与模型组比较,电针组大鼠机械痛和热敏痛阈值明显增加(P<0.05);与电针组和电针复合AV-shCTRL组比较,电针复合AV-shGATA4 RNA组大鼠机械痛和热敏痛阈值明显减小(P<0.05)。与假手术组比较,模型组大鼠脊髓腺苷A1受体和GATA4蛋白表达量明显增加(P<0.05)。与模型组比较,电针组和电针复合AV-shCTRL大鼠脊髓腺苷A1受体和GATA4蛋白表达量明显增加(P<0.05)。与电针组和电针复合AV-shCTRL组比较,电针复合AV-shGATA4 RNA组大鼠脊髓腺苷A1受体和GATA4蛋白表达量明显减少(P<0.05)。结论:电针可以增加CCI大鼠脊髓�Objective To investigate the effect of electroacupuncture(EA)on pain behaviors and expression of spinal transcription factor GATA-binding Protein 4(GATA4)and adenosine A1 receptor in neuropathic pain rats,so as to explore its mechanism underlying pain relief.Methods The present study includes 2 parts.In the first part,18 SD rats were randomly divided into control,adenovirus short-hairpin interference RNA for GATA4(AV-shGATA4 RNA)and adenovirus empty vector(AV-control short-hairpin RNA,AV-shCTRL)groups,with 6 rats in each group.The expression of GATA4 protein in the lumbar spinal cord(L4—L6)was detected to evaluate the transfection efficiency of AV-shGATA4 RNA(silencing GATA4 expression).In the second part,thirty SD rats were randomly divided into 5 groups,namely sham operation,CCI model,EA,EA+AV-shGATA4 RNA,and EA+AV-shCTRL groups,with 6 rats in each group.The neuropathic pain model was established by chronic constriction injury(CCI)of the right sciatic nerve.On the 7 th day following modeling,EA was applied to the right"Zusanli"(ST36)and"Taichong"(LR3)(1 mA,2 Hz/100 Hz)for 30 min.Rats of the EA+AV-shGATA4 RNA and EA+AV-shCTRL groups received intrathecal injection of AV-shGATA4 RNA and AV-shCTRL(1×1011 PFU/mL,10μL)at the spinal L4—L6 segments,separately,48 h before EA intervention.The mechanical pain threshold and thermal pain threshold of the affected limb were detected before molding,7 days following molding and 60 min after EA.The expressions of adenosine A1 receptor and GATA4 protein in the spinal cord(L4—L6)were detected by Western blot.Results Outcomes of the first part showed that compared with the control group,no significant changes were found in the mechanical and thermal pain thresholds in both AV-shCTRL and AV-shGATA4 RNA groups and in the expression of spinal GATA4 protein of the AV-shCTRL group(P>0.05).The expression of spinal GATA4 protein of the AV-shGATA4 RNA group was significantly lower than that of the AV-shCTRL group(P<0.05).In the second part of the study,before CCI modeling,there we

关 键 词：神经病理性痛 电针镇痛 腺苷A1受体 GATA4转录因子 

分 类 号：R245.9+7[医药卫生—针灸推拿学]