[1]范俊驰,宋建东,唐偲,等.电针夹脊穴调控脊髓损伤后炎症反应的作用机制研究[J].中医正骨,2020,32(05):1-5.
　FAN Junchi,SONG Jiandong,TANG Cai,et al.Study on mechanism of action of electroacupuncture at Point JIAJI(EX-B2)for regulating inflammatory reaction after spinal cord injury[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2020,32(05):1-5.
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电针夹脊穴调控脊髓损伤后炎症反应的作用机制研究()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第32卷
期数:
2020年05期
页码:
1-5
栏目:
基础研究
出版日期:
2020-05-20

文章信息/Info

Title:
Study on mechanism of action of electroacupuncture at Point JIAJI(EX-B2)for regulating inflammatory reaction after spinal cord injury
作者:
范俊驰; 宋建东; 唐偲; 汪今朝
(湖北省中西医结合医院,湖北 武汉 430015)
Author(s):
FAN Junchi; SONG Jiandong; TANG Cai; WANG Jinzhao
Hubei Combinational Hospital of Traditional Chinese Medicine and Western Medicine,Wuhan 430015,Hubei,China
关键词:
穴; 夹脊;  电针;  脊髓损伤;  大鼠;  炎症;  白细胞介素-1β;  白细胞介素-18;  NLRP3炎症小体
Keywords:
point ex-b2(JIAJI);  electroacupuncture;  spinal cord injuries;  rats;  inflammation;  interleukin-1beta;  interleukin-18;  NLRP3 inflammasome
摘要:
目的:探讨电针夹脊穴调控脊髓损伤后炎症反应的作用机制。方法:3月龄雌性SD大鼠48只,随机分为假手术组、模型组和电针组,每组16只。行椎板切除显露脊髓后,假手术组对脊髓不做处理,模型组和电针组用NYU脊髓打击器击打脊髓,建立脊髓损伤模型。造模成功后,电针组用电针刺激大鼠L1～L6双侧夹脊穴,每次20 min,每日早晚各1次,共干预3 d; 其余2组大鼠不做特殊干预。干预3 d后处理标本,采用尼氏染色法观察各组大鼠脊髓神经细胞形态,免疫组化法和Western blot法检测各组大鼠脊髓组织中NLRP3炎症小体的表达,ELISA法检测各组大鼠血清中白细胞介素(interleukin,IL)-1β、IL-18的含量。结果:①脊髓神经细胞形态。假手术组大鼠脊髓神经细胞形态完整,模型组大鼠脊髓神经细胞形态不规则,电针组大鼠脊髓神经细胞形态较模型组有所改善。②脊髓组织中NLRP3炎症小体的表达。免疫组化检测结果显示,NLRP3炎症小体主要在脊髓神经细胞胞浆中表达,呈棕褐色; 3组大鼠脊髓神经细胞中NLRP3炎症小体阳性相对表达量总体比较,差异有统计学意义[假手术组(1.000±0.000),模型组(1.599±0.009),电针组(1.294±0.007); F=1 912.233,P=0.000]; 电针组和模型组均高于假手术组(P=0.000,P=0.000),模型组高于电针组(P=0.000)。Western blot检测结果显示,3组大鼠脊髓组织中NLRP3炎症小体蛋白相对表达量总体比较,差异有统计学意义[假手术组(1.000±0.000),模型组(1.944±0.076),电针组(1.469±0.062); F=69.240,P=0.000]; 电针组和模型组均高于假手术组(P=0.000,P=0.000),模型组高于电针组(P=0.000)。③血清中IL-1β、IL-18含量。3组大鼠血清中IL-1β、IL-18含量总体比较,组间差异均有统计学意义[IL-1β:假手术组(3.500±0.423)pg·mL-1,模型组(24.500±1.180)pg·mL-1,电针组(13.000±0.886)pg·mL-1,F=140.742,P=0.000; IL-18:假手术组(4.750±0.675)pg·mL-1,模型组(28.375±1.879)pgmL-1,电针组(14.750±1.278)pg·mL-1,F=75.033,P=0.000]; 电针组和模型组均高于假手术组(P=0.000,P=0.000; P=0.000,P=0.000),模型组高于电针组(P=0.000,P=0.000)。结论:电针夹脊穴可抑制脊髓损伤大鼠脊髓组织中NLRP3炎症小体的表达,减少血清中炎症因子IL-1β和IL-18的含量,这可能是该疗法调控脊髓损伤后炎症发应、减轻脊髓神经细胞损伤的机制。
Abstract:
Objective:To explore the mechanism of action of electroacupuncture at Point JIAJI(EX-B2)for regulating inflammatory reaction after spinal cord injury(SCI).Methods:Forty-eight 3-month-old female SD rats were randomly divided into sham-operated group,model group and electroacupuncture group,16 cases in each group.The laminectomies were performed on rats in the 3 groups to expose their spinal cords.The spinal cords of rats in sham-operated group were not given any intervention; while the spinal cords of rats in model group and electroacupuncture group were hit by using NYU impactor to build the SCI models.After successful modeling,the rats in electroacupuncture group were stimulated with electroacupuncture at bilateral Point JIAJI(EX-B2)from L1 to L6,20 minutes at a time,twice a day in the morning and evening respectively for consecutive 3 days.No special intervention was performed on the rats in the other 2 groups.After 3-day intervention,the spinal nerve cell morphology of rats in the 3 groups were observed by using Nissl’s staining method,the expressions of NLRP3 inflammasomes in spinal cord tissues were detected by using immunohistochemical method and Western blot method,and the serum contents of interleukin(IL)-1β and IL-18 were detected by using ELISA method.Results:The spinal nerve cell morphology were complete in rats of sham-operated group and irregular in rats of model group and were improved in rats of electroacupuncture group compared to model group.The immunohistochemical staining results showed that NLRP3 inflammasomes were mainly expressed in cytoplasm of spinal nerve cells and presented with tawny staining.There was statistical difference in relative levels of positive expression of NLRP3 inflammasomes in spinal nerve cells of rats between the 3 groups in general(sham-operated group:1.000+/-0.000,model group:1.599+/-0.009,electroacupuncture group:1.294+/-0.007; F=1 912.233,P=0.000).The relative levels of positive expression of NLRP3 inflammasomes in spinal nerve cells were higher in electroacupuncture group and model group compared to sham-operated group,and were higher in model group compared to electroacupuncture group(P=0.000,P=0.000; P=0.000).The results of Western blot assay showed that there was statistical difference in the relative levels of protein expression of NLRP3 inflammasomes in spinal cord tissues of rats between the 3 groups in general(sham-operated group:1.000+/-0.000; model group:1.944+/-0.076; electroacupuncture group:1.469+/-0.062; F=69.240,P=0.000).The relative levels of protein expression of NLRP3 inflammasomes in spinal cord tissues of rats were higher in electroacupuncture group and model group compared to sham-operated group,and were higher in model group compared to electroacupuncture group(P=0.000,P=0.000; P=0.000).There was statistical difference in serum contents of IL-1β and IL-18 between the 3 groups in general(sham-operated group:3.500+/-0.423 pg/mL,model group:24.500+/-1.180 pg/mL,electroacupuncture group:13.000+/-0.886 pg/mL,F=140.742,P=0.000; sham-operated group:4.750+/-0.675 pg/mL,model group:28.375+/-1.879 pg/mL,electroacupuncture group:14.750+/-1.278 pg/mL,F=75.033,P=0.000).The serum contents of IL-1β and IL-18 were higher in electroacupuncture group and model group compared to sham-operated group,and were higher in model group compared to electroacupuncture group(P=0.000,P=0.000; P=0.000,P=0.000; P=0.000,P=0.000).Conclusion:Electroacupuncture at Point JIAJI(EX-B2)can inhibit the expression of NLRP3 inflammasomes in spinal cord tissues and decrease the serum contents of IL-1β and IL-18 in rats with spinal cord injuries,which may be its mechanism of action in regulating inflammatory reaction and lessening spinal nerve cell injuries after SCI.

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备注/Memo

备注/Memo:
(收稿日期:2020-01-23 本文编辑:杨雅)通讯作者:汪今朝 E-mail:wangjinzhao0326@163.com
更新日期/Last Update: 2020-08-15