电针预处理对慢性炎性痛大鼠脊髓背角组织中mTOR-自噬通路活性的影响  

Effect of electroacupuncture pretreatment on mTOR-autophagy pathway activity in spinal dorsal horn tissues of rats with chronic inflammatory pain

作　　者：吕丹[1] 栾静[1] 杨艳梅[1] 贺永进[1] LYU Dan;LUAN Jing;YANG Yanmei;HE Yongjin(Tianjin First Genter Hospital,Tianjin 300192,China)

机构地区：[1]天津市第一中心医院,天津300192

出　　处：《山东医药》2021年第17期29-32,共4页Shandong Medical Journal

基　　金：天津市卫生健康委、天津市中医药管理局中医、中西医结合科研课题一般项目(2019101);天津市第一中心医院春系列项目(2019CF26);天津市第一中心医院春系列项目(2019CF33)。

摘　　要：目的观察电针预处理对慢性炎性痛大鼠脊髓背角组织中哺乳动物雷帕霉素靶蛋白(mTOR)—自噬通路活性的影响。方法取健康成年雄性SD大鼠32只,按随机数字表法分为对照组(C组)、炎性痛组(IP组)、电针组(EA组)、非穴位电针组(NE组)各8只。IP组、EA组、NE组大鼠左侧后肢足底皮下注射蜜蜂毒溶液制备慢性炎性痛模型,C组同部位注射等体积无菌生理盐水;EA组造模前3 d每天给予左侧阳陵泉、足三里电针刺激,电针结束后1 h造模;NE组左侧阳陵泉、足三里旁开5 mm给予电针刺激。造模后2 h,测定机械刺激缩足阈(MWT)和热刺激缩足潜伏期(TWL);痛阈测定结束后处死大鼠,取脊髓背角组织,采用Westernblotting法检测自噬相关蛋白(LC3Ⅱ、Beclin-1、p62)及mTOR相关信号通路蛋白(mTOR、p-mTOR、S6K、p-S6K)。结果与C组比较,IP组和NE组MWT降低、TWL缩短(P均<0.01),EA组各指标差异无统计学意义;与IP组比较,EA组MWT升高、TWL延长(P均<0.01),NE组各指标差异无统计学意义。与C组比较,其余各组脊髓背角组织中LC3Ⅱ、Beclin-1和p62表达上调,IP组和NE组脊髓背角组织中p-mTOR、p-S6K表达上调(P均<0.05);与IP组比较,EA组脊髓背角组织中LC3Ⅱ和Beclin-1表达上调而p62、p-mTOR、p-S6K表达下调(P均<0.05),NE组各指标差异无统计学意义。结论电针预处理能激活mTOR信号通路调节脊髓背角的自噬状态以缓解大鼠慢性炎性痛。Objective To evaluate the effect of electroacupuncture pretreatment on mammalian target of rapamycin(mTOR)-autophagy signaling pathway activity in spinal dorsal horn of rats with chronic inflammatory pain.Methods Thirty-two adult male Sprague-Dawley rats were randomly divided into the control group(group C),inflammatory pain group(IP group),electroacupuncture group(EA group),and non-acupoint electroacupuncture group(NE group),with 8 in each.Chronic inflammatory pain models of rats were made by plantar subcutaneous injection of 50μL bee venom into rats´left hind limbs in the IP group,EA group and NE group,while the same volume of 0.9%sterile saline instead of bee vemon was injected into the same position in the group C;in the EA group,electroacupuncture was used to stimulate the left side of Yanglingquan(GB34)and Zusanli(ST36)every day for 3 days before modeling,and then the model was created 1 h after electroacupuncture pretreatment;in the NE group,the points 5 mm nearby rats´left Yanglingquan(GB34)and Zusanli(ST36)were pretreated with electroacupuncture.The mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were measured at 2 h after modeling;rats were sacrificed after the pain threshold measurement and spinal dorsal horn tissues were isolated,and we detected the expression levels of autophagy-related protein(microtubule-associated protein light chain type 3Ⅱ(LC3Ⅱ),Beclin-1 and p62)and mTOR-related signaling pathway proteins[mTOR,phosphorylated mTOR(p-mTOR),S6 kinase(S6K)and phosphorylated-S6K(p-S6K)]by Western blotting.Results Compared with those in the group C,MWT decreased and TWL became shorter both in the IP group and NE group(both P<0.01),while there was no significant statistical difference in the indicators of the EA group.Compared with those in the IP group,MWT increased and TWL became prolonged in the EA group(both P<0.01),while there was no statistically significant difference for the indicators in the NE group.In contrast with those in the group C,the expression levels of L

关 键 词：自噬 哺乳动物雷帕霉素靶蛋白 电针 炎性痛 脊髓背角 大鼠 

分 类 号：R338.2[医药卫生—人体生理学]