电针对急性术后痛大鼠背根神经节PD-L1/PD-1-SHP-1信号通路的影响  

Effect of electroacupuncture on PD-L1/PD-1-SHP-1 signaling pathway in dorsal root ganglia of rats with acute postoperative pain

作　　者：邓达玲 徐锋 陈向东 Deng Daling;Xu Feng;Chen Xiangdong(Department of Anesthesiology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China)

机构地区：[1]华中科技大学同济医学院附属协和医院麻醉科,武汉430022

出　　处：《中华麻醉学杂志》2023年第9期1108-1112,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82071251);国家重点研发计划(2018YFC2001802);湖北省重点研发计划(2021BCA145)。

摘　　要：目的评价电针对急性术后痛大鼠背根神经节(DRG)程序性细胞死亡配体1(PD-L1)/程序性细胞死亡受体(PD-1)/含Src-同源区2蛋白酪氨酸磷酸酶-1(SHP-1)信号通路的影响。方法SPF级雄性SD大鼠39只,体质量220~250 g,6~8周龄,采用随机数字表法分为3组(n=13):对照组(C组)、腹部手术组(S组)和腹部手术+电针组(S+EA组)。S组和S+EA组在异氟烷麻醉下行腹部手术。S+EA组于术毕即刻和术后2 h时,选取双侧足三里和三阴交穴进行电针刺激30 min,频率为10 Hz连续波,电流为1 mA。每组取7只大鼠,分别于造模前1 d(T_(0))和造模后3、5、7、9、11、24 h时(T_(1~6)),依次测定机械缩足反应阈(MWT)、机械缩腹反应阈(ACT)、热缩足潜伏期(TWL)和冷缩足潜伏期(CWL)。于造模后5 h每组处死6只大鼠,取出T_(12)~L_(4) DRG,分别采用Western blot法检测IL-6、PD-L1、PD-1和SHP-1的表达,采用免疫荧光法观察PD-L1在各类神经元的表达情况。结果3组不同时点TWL和CWL比较差异无统计学意义(P>0.05)。与C组比较,S组T_(1~5)时MWT降低,T_(1~6)时ACT降低,DRG IL-6表达上调,PD-L1、PD-1和SHP-1表达下调(P<0.05);与S组比较,S+EA组T1,2时MWT和ACT升高,DRG IL-6表达下调,PD-L1、PD-1和SHP-1表达上调(P<0.05)。免疫荧光结果显示,PD-L1在小直径神经元(CGRP+神经元和IB4+神经元)和大直径神经元(NF200+神经元)上均存在表达,且主要表达在CGRP+神经元和IB4^(+)神经元上。3组间PD-L1在DRG各类神经元的表达比较差异无统计学意义(P>0.05)。结论电针减轻大鼠急性术后痛的机制可能与激活PD-L1/PD-1/SHP-1信号通路有关。Objective To evaluate the effect of electroacupuncture(EA)on programmed cell death ligand-1(PD-L1)/programmed cell death receptor-1(PD-1)/Src-homology region two domain-containing phosphatase-1(SHP-1)signaling pathway in the dorsal root ganglia(DRG)of rats with acute postoperative pain.Methods Thirty-nine SPF male Sprague-Dawley rats,weighing 220-250 g,aged 6-8 weeks,were divided into 3 groups(n=13 each)using a random number table method:control group(group C),abdominal surgery group(group S)and abdominal surgery+EA group(group S+EA).Group S and group S+EA underwent abdominal surgery under isoflurane anesthesia.In S+EA group,both Zusanli and Sanyinjiao acupoints were selected and stimulated for 30 min with a frequency of 10 Hz continuous wave and a current of 1 mA starting from the end of operation and 2 h after operation.Seven rats were selected from each group for measurement of the mechanical paw withdrawal threshold(MWT),abdominal contraction threshold(ACT),thermal paw withdrawal latency(TWL)and cold paw withdrawal latency(CWL)at 1 day before developing the model(T_(0))and 3,5,7,9,11 and 24 h after developing the model(T_(1-6)).Six rats in each group were sacrificed at 5 h after the model was prepared,and T_(12)-L_(4) DRG was removed for determination of the expression of interleukin-6(IL-6),PD-L1,PD-1 and SHP-1(by Western blot)and expression of PD-L1 in various neurons(by immunofluorescence).Results There was no significant difference in TWL and CWL between the three groups at different time points(P>0.05).Compared with group C,MWT at T_(1-5) and ACT at T_(1-6)were significantly decreased,and the expression of IL-6 in DRG was up-regulated,and the expression of PD-L1,PD-1 and SHP-1 in DRG was down-regulated in group S(P<0.05).Compared with group S,MWT and ACT were significantly increased at T_(1,2),the expression of IL-6 in DRG was down-regulated,and the expression of PD-L1,PD-1 and SHP-1 in DRG was up-regulated in group S+EA(P<0.05).The results of immunofluorescence showed that PD-L1 was expressed on both sm

关 键 词：电针 疼痛 手术后 神经节 B7-H1抗原 程序性细胞死亡受体1 蛋白酪氨酸磷酸酶 非受体6型 

分 类 号：R245[医药卫生—针灸推拿学]