Article(id=1210676790213866430, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1210676785113600955, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.2021.05.06, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1611158400000, receivedDateStr=2021-01-21, revisedDate=1615132800000, revisedDateStr=2021-03-08, acceptedDate=null, acceptedDateStr=null, onlineDate=1766577440073, onlineDateStr=2025-12-24, pubDate=1622131200000, pubDateStr=2021-05-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1766577440073, onlineIssueDateStr=2025-12-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1766577440073, creator=13701087609, updateTime=1766577440073, updator=13701087609, issue=Issue{id=1210676785113600955, tenantId=1146029695717560320, journalId=1189873630562394117, year='2021', volume='46', issue='5', pageStart='425', pageEnd='530', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1766577438858, creator=13701087609, updateTime=1766718730270, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1211269404306838321, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1210676785113600955, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1211269404306838322, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1210676785113600955, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=462, endPage=467, ext={EN=ArticleExt(id=1210676790549410772, articleId=1210676790213866430, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=Analgesic effect of low-intensity exercise induced by interleukin-4 on chronic constriction injury of sciatic nerve in mice, columnId=1190310110212751762, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Basic Research, runingTitle=null, highlight=null, articleAbstract=

Objective To explore the analgesic effect of low-intensity exercise induced by interleukin (IL)-4 on chronic constriction injury of the sciatic nerve (CCI) mice. Methods A total of 48 male C57/BL6 mice were randomized into 6 groups(n=8): sham/sedentary group (SS group), sham/normal saline/exercise group (SNE group), sham/anti-IL-4/exercise group (SAE group), CCI/sedentary group (CS group), CCI/normal saline/exercise group (CNE group), CCI/anti-IL-4/exercise group (CAE group). After inducing CCI, mice were given low-intensity exercise from 1 to 15 days. For the SAE group and the CAE group, mice received 50 μg of IL-4 receptor blocker (IL-4Ra) 30 min before exercise through intraperitoneal injection. Behavioural tests were performed on the 3rd day (T1), 7th day (T2), 11th day (T3) and 15th day (T4) to analyze the difference of hyperalgesia. On the 15th day after CCI induction, the injured sciatic nerve and lumbar spinal cord (L1-L6) of all mice were collected. The levels of IL-4, IL-1Ra, IL-5, TNF-α, IL-1β, brain-derived neurotrophic factor (BDNF) and β-nerve growth factor (β-NGF) were detected by ELISA. Results The paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) values of the CNE group at T3 and T4 were higher than those of the CS group and CAE group (P<0.05); the IL-4 levels of the sciatic nerve and lumbar spinal cord in CNE group and CAE group were higher than those of CS group (P<0.05); the IL-1Ra and IL-5 levels of the sciatic nerve and lumbar spinal cord in CNE group were higher than those of CS group and CAE group (P<0.05); the TNF-α, IL-1β, BDNF and β-NGF levels of the sciatic nerve and lumbar spinal cord of CNE group were lower than those of CS group and CAE group (P<0.05). Conclusion Low-intensity exercise exerts the analgesic effect on mice with chronic constriction injury of sciatic nerve through IL-4.

, correspAuthors=Pei Wang, authorNote=null, correspAuthorsNote=
*E-mail:
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目的 探讨低强度运动能否通过白细胞介素(IL)-4介导对慢性坐骨神经缩窄性损伤(CCI)小鼠发挥镇痛作用。方法 雄性C57/BL6健康小鼠48只,采用随机数字表法分为6组(n=8):假手术/静坐组(SS组)、假手术/生理盐水/运动(SNE组)、假手术/抗IL-4/运动(SAE组)、CCI/静坐组(CS组)、CCI/生理盐水/运动组(CNE组)、CCI/抗IL-4/运动组(CAE组)。制备CCI模型,造模后第1—15天行低强度运动,SAE组和CAE组小鼠运动前30 min腹腔注射IL-4受体拮抗剂(IL-4Ra)50 μg。造模后第3(T1)、7(T2)、11(T3)、15(T4)天进行行为学测试以分析小鼠痛觉过敏的差异。造模后第15天取小鼠损伤侧坐骨神经和腰段脊髓(L1~L6),采用ELISA法检测IL-4、IL-1Ra、IL-5、肿瘤坏死因子-α(TNF-α)、IL-1β、脑源性神经营养因子(BDNF)、β-神经生长因子(β-NGF)的表达水平。结果 CNE组造模后T3和T4时的机械刺激缩足反射阈值(PWMT)和热刺激缩足反射潜伏期(PWTL)均高于CS组和CAE组(P<0.05);CNE组和CAE组坐骨神经和腰段脊髓的IL-4水平均高于CS组,差异有统计学意义(P<0.05);CNE组坐骨神经和腰段脊髓的IL-1Ra、IL-5水平均高于CS组和CAE组,TNF-α、IL-1β、BDNF、β-NGF水平均低于CS组和CAE组,差异有统计学意义(P<0.05)。结论 低强度运动可能通过IL-4介导对CCI小鼠产生镇痛作用。

, correspAuthors=汪佩, authorNote=null, correspAuthorsNote=
汪佩,E-mail:
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方平,硕士研究生,副主任医师,主要从事神经病理性疼痛研究

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J Cell Biochem, 2019, 120(8): 12604-12617., articleTitle=Docosahexaenoic acid enhances M2 macrophage polarization via the p38 signaling pathway and autophagy, refAbstract=null), Reference(id=1210676798094963033, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, doi=null, pmid=null, pmcid=null, year=2018, volume=66, issue=8, pageStart=1775, pageEnd=1787, url=null, language=null, rfNumber=[24], rfOrder=27, authorNames=Okutani H, Yamanaka H, Kobayashi K, journalName=Glia, refType=null, unstructuredReference=Okutani H, Yamanaka H, Kobayashi K, et al. Recombinant interleukin-4 alleviates mechanical allodynia via injury-induced interleukin-4 receptor alpha in spinal microglia in a rat model of neuropathic pain[J]. Glia, 2018, 66(8): 1775-1787., articleTitle=Recombinant interleukin-4 alleviates mechanical allodynia via injury-induced interleukin-4 receptor alpha in spinal microglia in a rat model of neuropathic pain, refAbstract=null)], funds=[Fund(id=1210676795452551412, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, awardId=202003N4246, language=EN, fundingSource=Natural Science Foundation of Ningbo City(202003N4246), fundOrder=null, country=null), Fund(id=1210676795540631799, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, awardId=202003N4246, language=CN, fundingSource=宁波市自然科学基金(202003N4246), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1210676791577014281, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, xref=1, ext=[AuthorCompanyExt(id=1210676791585402890, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, companyId=1210676791577014281, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1Department of Anesthesiology, Lihuili Hospital, Ningbo Medical Center, Ningbo, Zhejiang 315000, China), AuthorCompanyExt(id=1210676791593791500, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, companyId=1210676791577014281, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1浙江省宁波市医疗中心李惠利医院麻醉科,浙江 宁波 315000)]), AuthorCompany(id=1210676791707037714, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, xref=2, ext=[AuthorCompanyExt(id=1210676791715426324, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, companyId=1210676791707037714, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2Department of Operating Room, Lihuili Hospital, Ningbo Medical Center, Ningbo, Zhejiang 315000, China), AuthorCompanyExt(id=1210676791719620629, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, companyId=1210676791707037714, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2浙江省宁波市医疗中心李惠利医院手术室,浙江 宁波 315000)])], figs=[ArticleFig(id=1210676794680799441, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, language=EN, label=Fig.1, caption=Processing flow chart of mice in each group, figureFileSmall=opgHSVXRx8kW7Vp2J9B7HQ==, figureFileBig=XN1uJb6tqoA+9tFya5QZvw==, tableContent=null), ArticleFig(id=1210676794752102612, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, language=CN, label=图1, caption=各组小鼠处理流程

SS. 假手术/静坐;SNE. 假手术/生理盐水/运动;SAE. 假手术/抗IL-4/运动;CCI. 慢性坐骨神经缩窄性损伤;CS. CCI/静坐;CNE. CCI/生理盐水/运动;CAE. CCI/抗IL-4/运动;PWMT. 机械刺激缩足反射阈值;PWTL. 热刺激缩足反射潜伏期;ip. 腹腔注射;IL. 白细胞介素;ELISA. 酶联免疫吸附试验

, figureFileSmall=opgHSVXRx8kW7Vp2J9B7HQ==, figureFileBig=XN1uJb6tqoA+9tFya5QZvw==, tableContent=null), ArticleFig(id=1210676794995372255, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, language=EN, label=Fig.2, caption=Comparison of cytokine levels in sciatic nerve and lumbar spinal cord (L1-L6) of mice in each group (n=8), figureFileSmall=bPc4WUJv1BVZbqW0NHKHgw==, figureFileBig=6nZTFQCJaMm763Xr/jJVaA==, tableContent=null), ArticleFig(id=1210676795066675427, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, language=CN, label=图2, caption=各组小鼠坐骨神经和腰段脊髓(L1~L6)细胞因子水平比较(n=8)

IL. 白细胞介素;TNF-α. 肿瘤坏死因子-α;BDNF. 脑源性神经营养因子;β-NGF. 神经生长因子-β;A—G. 各组小鼠坐骨神经IL-4、IL-1Ra、IL-5、TNF-α、IL-1β、BDNF、β-NGF水平比较;H—N. 各组小鼠腰段脊髓(L1~L6)IL-4、IL-1Ra、IL-5、TNF-α、IL-1β、BDNF、β-NGF水平比较;与SS组比较,(1)P<0.05;与SNE组比较,(2)P<0.05;与SAE组比较,(3)P<0.05;与CS组比较,(4)P<0.05;与CNE组比较,(5)P<0.05。

, figureFileSmall=bPc4WUJv1BVZbqW0NHKHgw==, figureFileBig=6nZTFQCJaMm763Xr/jJVaA==, tableContent=null), ArticleFig(id=1210676795196698859, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, language=EN, label=Tab.1, caption=

Comparison of PWMT and PWTL values of mice in each group at different time points ($\bar{x}±s$, n=8)

, figureFileSmall=null, figureFileBig=null, tableContent=
指标SS组SNE组SAE组CS组CNE组CAE组FP
PWMT (g)
T03.0±0.13.0±0.32.8±0.23.0±0.33.1±0.23.0±0.31.0410.406
T13.1±0.43.1±0.63.1±0.31.7±0.2(1)(2)(3)(6)1.9±0.2(1)(2)(3)(6)1.6±0.3(1)(2)(3)(5)(6)21.8800.000
T23.0±0.42.9±0.62.8±0.51.3±0.1(1)(2)(3)(6)1.8±0.3(1)(2)(3)(6)1.3±0.2(1)(2)(3)(6)29.5930.000
T32.8±0.32.7±0.32.5±0.31.3±0.1(1)(2)(3)(5)(6)2.0±0.1(1)(2)(3)(4)(6)1.2±0.1(1)(2)(3)(5)(6)48.8260.000
T42.9±0.22.8±0.22.6±0.41.5±0.1(1)(2)(3)(5)(6)2.2±0.2(1)(2)(3)(4)(6)1.4±0.2(1)(2)(3)(5)(6)47.4700.000
PWTL (s)
T07.6±1.09.3±1.39.2±1.38.7±0.79.1±1.39.0±1.42.1590.077
T16.5±1.17.6±1.07.8±1.52.7±0.8(1)(2)(3)(6)3.5±1.1(1)(2)(3)(6)2.6±0.7(1)(2)(3)(6)37.5340.000
T26.8±1.87.3±1.77.1±1.91.6±0.9(1)(2)(3)(6)3.3±0.7(1)(2)(3)(6)1.7±0.7(1)(2)(3)(6)29.0460.000
T37.3±1.77.5±2.17.1±1.52.2±0.7(1)(2)(3)(5)(6)4.8±0.9(1)(2)(3)(4)(6)2.2±1.0(1)(2)(3)(5)(6)23.7420.000
T46.2±0.77.1±1.17.5±1.61.8±0.7(1)(2)(3)(5)(6)4.4±0.9(1)(2)(3)(4)(6)1.5±0.7(1)(2)(3)(5)(6)50.3870.00
), ArticleFig(id=1210676795326722287, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1210676790213866430, language=CN, label=表1, caption=

各组小鼠不同时间点PWMT、PWTL比较($\bar{x}±s$,n=8)

, figureFileSmall=null, figureFileBig=null, tableContent=
指标SS组SNE组SAE组CS组CNE组CAE组FP
PWMT (g)
T03.0±0.13.0±0.32.8±0.23.0±0.33.1±0.23.0±0.31.0410.406
T13.1±0.43.1±0.63.1±0.31.7±0.2(1)(2)(3)(6)1.9±0.2(1)(2)(3)(6)1.6±0.3(1)(2)(3)(5)(6)21.8800.000
T23.0±0.42.9±0.62.8±0.51.3±0.1(1)(2)(3)(6)1.8±0.3(1)(2)(3)(6)1.3±0.2(1)(2)(3)(6)29.5930.000
T32.8±0.32.7±0.32.5±0.31.3±0.1(1)(2)(3)(5)(6)2.0±0.1(1)(2)(3)(4)(6)1.2±0.1(1)(2)(3)(5)(6)48.8260.000
T42.9±0.22.8±0.22.6±0.41.5±0.1(1)(2)(3)(5)(6)2.2±0.2(1)(2)(3)(4)(6)1.4±0.2(1)(2)(3)(5)(6)47.4700.000
PWTL (s)
T07.6±1.09.3±1.39.2±1.38.7±0.79.1±1.39.0±1.42.1590.077
T16.5±1.17.6±1.07.8±1.52.7±0.8(1)(2)(3)(6)3.5±1.1(1)(2)(3)(6)2.6±0.7(1)(2)(3)(6)37.5340.000
T26.8±1.87.3±1.77.1±1.91.6±0.9(1)(2)(3)(6)3.3±0.7(1)(2)(3)(6)1.7±0.7(1)(2)(3)(6)29.0460.000
T37.3±1.77.5±2.17.1±1.52.2±0.7(1)(2)(3)(5)(6)4.8±0.9(1)(2)(3)(4)(6)2.2±1.0(1)(2)(3)(5)(6)23.7420.000
T46.2±0.77.1±1.17.5±1.61.8±0.7(1)(2)(3)(5)(6)4.4±0.9(1)(2)(3)(4)(6)1.5±0.7(1)(2)(3)(5)(6)50.3870.00
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白细胞介素-4介导低强度运动对慢性坐骨神经缩窄性损伤小鼠的镇痛作用
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方平 1 , 王静玉 2 , 许周亮 1 , 孙刚强 1 , 荀玉月 1 , 汪佩 1, *
解放军医学杂志 | 基础研究 2021,46(5): 462-467
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解放军医学杂志 | 基础研究 2021, 46(5): 462-467
白细胞介素-4介导低强度运动对慢性坐骨神经缩窄性损伤小鼠的镇痛作用
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方平1, 王静玉2, 许周亮1, 孙刚强1, 荀玉月1, 汪佩1, *
作者信息
  • 1浙江省宁波市医疗中心李惠利医院麻醉科,浙江 宁波 315000
  • 2浙江省宁波市医疗中心李惠利医院手术室,浙江 宁波 315000
  • 方平,硕士研究生,副主任医师,主要从事神经病理性疼痛研究

通讯作者:

汪佩,E-mail:
Analgesic effect of low-intensity exercise induced by interleukin-4 on chronic constriction injury of sciatic nerve in mice
Ping Fang1, Jing-Yu Wang2, Zhou-Liang Xu1, Gang-Qiang Sun1, Yu-Yue Xun1, Pei Wang1, *
Affiliations
  • 1Department of Anesthesiology, Lihuili Hospital, Ningbo Medical Center, Ningbo, Zhejiang 315000, China
  • 2Department of Operating Room, Lihuili Hospital, Ningbo Medical Center, Ningbo, Zhejiang 315000, China
出版时间: 2021-05-28 doi: 10.11855/j.issn.0577-7402.2021.05.06
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目的 探讨低强度运动能否通过白细胞介素(IL)-4介导对慢性坐骨神经缩窄性损伤(CCI)小鼠发挥镇痛作用。方法 雄性C57/BL6健康小鼠48只,采用随机数字表法分为6组(n=8):假手术/静坐组(SS组)、假手术/生理盐水/运动(SNE组)、假手术/抗IL-4/运动(SAE组)、CCI/静坐组(CS组)、CCI/生理盐水/运动组(CNE组)、CCI/抗IL-4/运动组(CAE组)。制备CCI模型,造模后第1—15天行低强度运动,SAE组和CAE组小鼠运动前30 min腹腔注射IL-4受体拮抗剂(IL-4Ra)50 μg。造模后第3(T1)、7(T2)、11(T3)、15(T4)天进行行为学测试以分析小鼠痛觉过敏的差异。造模后第15天取小鼠损伤侧坐骨神经和腰段脊髓(L1~L6),采用ELISA法检测IL-4、IL-1Ra、IL-5、肿瘤坏死因子-α(TNF-α)、IL-1β、脑源性神经营养因子(BDNF)、β-神经生长因子(β-NGF)的表达水平。结果 CNE组造模后T3和T4时的机械刺激缩足反射阈值(PWMT)和热刺激缩足反射潜伏期(PWTL)均高于CS组和CAE组(P<0.05);CNE组和CAE组坐骨神经和腰段脊髓的IL-4水平均高于CS组,差异有统计学意义(P<0.05);CNE组坐骨神经和腰段脊髓的IL-1Ra、IL-5水平均高于CS组和CAE组,TNF-α、IL-1β、BDNF、β-NGF水平均低于CS组和CAE组,差异有统计学意义(P<0.05)。结论 低强度运动可能通过IL-4介导对CCI小鼠产生镇痛作用。

低强度运动  /  白细胞介素-4  /  镇痛  /  慢性坐骨神经缩窄性损伤  /  疼痛

Objective To explore the analgesic effect of low-intensity exercise induced by interleukin (IL)-4 on chronic constriction injury of the sciatic nerve (CCI) mice. Methods A total of 48 male C57/BL6 mice were randomized into 6 groups(n=8): sham/sedentary group (SS group), sham/normal saline/exercise group (SNE group), sham/anti-IL-4/exercise group (SAE group), CCI/sedentary group (CS group), CCI/normal saline/exercise group (CNE group), CCI/anti-IL-4/exercise group (CAE group). After inducing CCI, mice were given low-intensity exercise from 1 to 15 days. For the SAE group and the CAE group, mice received 50 μg of IL-4 receptor blocker (IL-4Ra) 30 min before exercise through intraperitoneal injection. Behavioural tests were performed on the 3rd day (T1), 7th day (T2), 11th day (T3) and 15th day (T4) to analyze the difference of hyperalgesia. On the 15th day after CCI induction, the injured sciatic nerve and lumbar spinal cord (L1-L6) of all mice were collected. The levels of IL-4, IL-1Ra, IL-5, TNF-α, IL-1β, brain-derived neurotrophic factor (BDNF) and β-nerve growth factor (β-NGF) were detected by ELISA. Results The paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) values of the CNE group at T3 and T4 were higher than those of the CS group and CAE group (P<0.05); the IL-4 levels of the sciatic nerve and lumbar spinal cord in CNE group and CAE group were higher than those of CS group (P<0.05); the IL-1Ra and IL-5 levels of the sciatic nerve and lumbar spinal cord in CNE group were higher than those of CS group and CAE group (P<0.05); the TNF-α, IL-1β, BDNF and β-NGF levels of the sciatic nerve and lumbar spinal cord of CNE group were lower than those of CS group and CAE group (P<0.05). Conclusion Low-intensity exercise exerts the analgesic effect on mice with chronic constriction injury of sciatic nerve through IL-4.

low-intensity exercise  /  interleukin-4  /  analgesic  /  chronic constriction injury of sciatic nerve  /  pain
方平, 王静玉, 许周亮, 孙刚强, 荀玉月, 汪佩. 白细胞介素-4介导低强度运动对慢性坐骨神经缩窄性损伤小鼠的镇痛作用. 解放军医学杂志, 2021 , 46 (5) : 462 -467 . DOI: 10.11855/j.issn.0577-7402.2021.05.06
Ping Fang, Jing-Yu Wang, Zhou-Liang Xu, Gang-Qiang Sun, Yu-Yue Xun, Pei Wang. Analgesic effect of low-intensity exercise induced by interleukin-4 on chronic constriction injury of sciatic nerve in mice[J]. Medical Journal of Chinese People’s Liberation Army, 2021 , 46 (5) : 462 -467 . DOI: 10.11855/j.issn.0577-7402.2021.05.06
慢性坐骨神经缩窄性损伤(chronic constriction injury of the sciatic nerve,CCI)引起的神经病理性疼痛在我国人群中发病率高达8.2%,严重影响患者的生活质量,甚至导致部分患者功能丧失、残疾[1-3]。低强度运动指运动时所测得的心率≤40%最大运动心率的运动强度。有研究证实规律运动具有一定的镇痛作用[4-5],是临床实践指南推荐的一项减轻慢性疼痛和促进神经功能恢复的策略。Lopes等[6]发现,进行规律的有氧运动可减轻疼痛,同时白细胞介素-4(interleukin-4,IL-4)水平较运动前有所升高;Kiguchi等[7]发现,将IL-4注射至小鼠神经损伤部位可有效减轻疼痛。本研究通过构建小鼠CCI模型,探讨低强度运动是否通过IL-4介导从而对神经病理性疼痛小鼠发挥镇痛作用。
小鼠IL-4受体拮抗剂(IL-4Ra,PZ0363-5MG)、抗神经生长因子单克隆抗体(N3279)以及IL-4、IL-1β ELISA试剂盒(RAB030、RAB0275)购自美国Sigma公司;小鼠抗脑源性神经营养因子(BDNF)单克隆抗体(JLC4037)以及IL-5、IL-1受体拮抗剂(IL-1Ra)、肿瘤坏死因子-α(TNF-α)ELISA试剂盒(JLC3600、JLC3582、JLC3924)购自上海晶抗生物工程有限公司;神经生长因子-β(nerve growth factor-β,β-NGF)ELISA试剂盒(96T)购自北京百奥莱博科技有限公司。小动物跑步机(SA101)购自江苏赛昂斯生物科技有限公司;电子机械痛阈测试仪(2390-5)、爪热痛测试仪(3730)购自美国IITC公司;纤毛机械刺激针购自上海玉研科学仪器有限公司;恒温手术台(Size1)购自美国Harvard公司;台式高速冷冻离心机(M1324R)购自深圳市瑞沃德生命科技有限公司。
健康雄性C57/BL6小鼠48只,8周龄,20~30 g,由宁波大学动物实验中心提供。小鼠于21~23℃和50%~80%湿度条件下饲养,12h /12 h光/暗循环,给予鼠类标准颗粒饲料(宁波大学实验动物中心提供)并自由饮水。采用随机数字表法分为6组(n=8):假手术/静坐组(SS组)、假手术/生理盐水/运动组(SNE组)、假手术/抗IL-4/运动组(SAE组)、CCI/静坐组(CS组)、CCI/生理盐水/运动组(CNE组)、CCI/抗IL-4/运动组(CAE组)。分组及各组处理详情如图1所示。实验过程符合国家和单位有关实验动物的管理和使用规定。
参照文献[8]的方法制备CCI小鼠模型:小鼠腹腔注射3%戊巴比妥钠0.04 mg/g麻醉后取侧卧位,消毒铺巾,切开右下肢股部皮肤,逐层分离直至完整暴露坐骨神经,于主干部位用4-0铬制羊肠线松扎4道间距为1 mm的松结扎,结扎强度以大腿产生1个小的短暂性抽搐为宜,结扎后逐层缝合切口观察至麻醉苏醒,假手术组小鼠仅暴露右侧坐骨神经但不结扎。通过测定并对比损伤侧与对侧机械刺激缩足反射阈值(paw withdrawal mechanical threshold,PWMT)剔除不合格样本,确定造模成功。
所有小鼠均以术前3 d(T0)的行为学测试结果作为基线值,造模后第1—15天运动组行低强度跑步机运动,造模后第3(T1)、7(T2)、11(T3)、15(T4)天进行行为学测试以分析小鼠痛觉过敏的差异。
将小鼠放入测试框内并置于网格面板上适应0.5 h,待小鼠安静后测定机械痛阈值。将纤毛机械刺激针固定于支架上,开启并调零测痛仪,钝性探针以恒定速度垂直向上刺激小鼠右足底正中皮肤,至出现舔足或缩足时记录针刺力度(g)即为PWMT,每次测定间隔至少5 min,相同的爪子重复测5次,除去最大值和最小值后取3次数据平均值作为小鼠后爪的PWMT值。
将小鼠放入测试框内适应0.5h,待小鼠安静后,设置爪热痛测试仪的红外热辐射强度为60%且单次加热刺激最长时间为20 s,以避免热辐射损伤动物足底组织影响后续实验的准确性。调整光源正对于小鼠右侧足底正中后开始计时,记录出现舔足或缩足反射时的持续光照时间即为PWTL。每次测定间隔至少5 min,相同的爪子重复测5次,除去最大值和最小值后取3次数据平均值作为小鼠后爪的PWTL值。
为明确IL-4介导运动对CCI小鼠的镇痛作用,SAE组、CAE组小鼠运动前30 min腹腔注射IL-4Ra 50 μg,SNE组、CNE组在相同时间点腹腔注射等量生理盐水。
造模后第1—15天,将SNE组、SAE组、CNE组、CAE组小鼠放置于跑步机上开始低强度有氧跑步机跑步,1次/d,30 min/次(无倾斜度,10 m/min,5 d/周)。
造模后第15天完成最后一次行为学测试后,腹腔注射3%戊巴比妥钠0.04 mg/g麻醉小鼠,断头处死,取损伤侧坐骨神经和腰段脊髓(L1~L6),采用ELISA法检测IL-4、IL-1Ra、IL-5、TNF-α、IL-1β、BDNF、β-NGF水平。
采用SPSS 24.0软件进行统计分析。符合正态分布且方差齐的计量资料以$\bar{x}±s$表示,多组间比较采用单因素方差分析,不同时间点比较采用双因素重复测量方差分析,进一步两两比较采用LSD事后检验。P<0.05为差异有统计学意义。
造模前各组小鼠PWMT、PWTL比较差异无统计学意义(P>0.05);SS组、SNE组和SAE组造模前后各时间点的PWMT、PWTL差异无统计学意义(P>0.05);CS组、CNE组和CAE组造模后各时间点的PWMT、PWTL低于造模前基线值,差异有统计学意义(P<0.05);CS组、CNE组和CAE组造模后PWMT、PWTL低于SS组、SNE组及SAE组,差异均有统计学意义(P<0.05);CNE组造模后T3和T4时的PWMT、PWTL高于CS组,而CAE组低于CNE组,差异有统计学意义(P<0.05)(表1)。
SS组、SNE组和SAE组坐骨神经和腰段脊髓的IL-4、IL-1Ra、IL-5、TNF-α、IL-1β、BDNF、β-NGF水平差异无统计学意义(P>0.05);CS组、CNE组和CAE组坐骨神经和腰段脊髓的IL-4、IL-1Ra、IL-5水平均低于SS组、SNE组和SAE组,TNF-α、IL-1β、BDNF、β-NGF水平高于SS组、SNE组和SAE组,差异有统计学意义(P<0.05);CNE组和CAE组坐骨神经和腰段脊髓的IL-4水平均高于CS组,差异有统计学意义(P<0.05);CNE组坐骨神经和腰段脊髓的IL-1Ra、IL-5水平均高于CS组和CAE组,差异有统计学意义(P<0.05);CNE组坐骨神经和腰段脊髓的TNF-α、IL-1β、BDNF、β-NGF水平低于CS组,而CAE组高于CNE组,差异有统计学意义(P<0.05);CS组与CAE组坐骨神经和腰段脊髓各细胞因子水平差异无统计学意义(P>0.05)(图2)。
坐骨神经损伤后损伤部位和脊髓内的神经免疫相互作用在神经病理性疼痛的发生与发展中起重要作用:神经损伤所释放的促炎细胞因子如IL-1β、TNF-α等在疼痛的产生中具有重要作用[9-10];而抗炎细胞因子如IL-5、IL-1Ra等可抑制炎症反应而减轻疼痛[11];与此同时,脊髓背角中的星形胶质细胞和小胶质细胞被激活,释放IL-1β、TNF-α、BDNF等促炎因子以及神经生长因子,上调背角神经元的兴奋性,从而导致痛觉过敏[12]。本研究中CS组、CNE组和CAE组小鼠各时间点机械刺激痛阈值和热痛觉过敏阈值均明显下降,表明CS组、CNE组和CAE组小鼠较SS组、SNE组、SAE组更易发生痛觉过敏;CS组、CNE组和CAE组小鼠损伤部位和腰段脊髓的促炎细胞因子和神经生长因子水平高于SS组、SNE组、SAE组,而抗炎因子水平低于SS组、SNE组、SAE组,提示神经损伤后外周损伤部位和脊髓的炎症反应加重,同时提示造模成功。CNE组在进行2周低强度运动后PWMT和PWTL较CS组和CAE组有所上升,但仍低于假手术组,提示CCI导致的神经病理性疼痛在低强度运动后有所缓解。
低强度运动是一种低成本、安全、有效的疼痛干预措施,深入探索其机制不仅有助于改善慢性疼痛的持续状态,还可为临床医师开具廉价、安全、便捷的非药物治疗处方提供依据。多项研究表明,规律运动不仅有利于缓解疼痛,还可促进神经保护与再生[13-14]。外周神经损伤后大量促炎因子释放导致机体局部痛阈下降,而规律的低强度运动可促进抗炎细胞因子释放以减轻炎症反应,从而发挥局部镇痛作用[15-16];与此同时,脊髓背角中的星型胶质细胞和小胶质细胞在神经损伤后被激活并释放BDNF以增强背角神经元的兴奋性,而规律运动后释放的抗炎因子可直接抑制BDNF和β-NGF的活性,脊髓背角神经元的兴奋性随之下降,导致机体痛阈上升[15-16]。本研究造模后经连续低强度运动后测得CNE组促炎因子IL-1β、TNF-α、β-NGF、BDNF水平低于CS组和CAE组,抗炎因子IL-5、IL-1Ra水平高于CS组和CAE组,提示规律低强度运动可有效减轻神经损伤部位和中枢的炎症反应,同时与CNE组机械刺激痛阈和热刺激痛阈上升的结果相互印证,表明运动可在局部和中枢同时发挥抗炎镇痛作用,与Harris等[17]的研究结果一致。
IL-4是巨噬细胞趋化因子,由Ⅱ型辅助T细胞(Th2细胞)分泌并在炎症反应中起重要作用[18-19]。近年来研究发现,在损伤部位或脊髓内注射IL-4可预防神经损伤引起的炎症和痛觉过敏,还可逆转神经损伤后脊髓胶质细胞的活化,且这种镇痛机制并非通过内源性阿片类递质及受体作用完成,而是与其抑制炎性因子的外周机制有关[18];相反,IL-4表达下调或其作用被拮抗可促进IL-1β、TNF-α等炎性介质的释放,从而加重疼痛[20-22]。Kiguchi等[7]研究发现,IL-4可有效缓解小鼠局部神经损伤部位的疼痛,其镇痛机制为:通过脂多糖下调M1型巨噬细胞特异性分子IL-1β、CC趋化因子配体-3和CD86 mRNA的表达[22];通过磷酸化信号传导及转录激活蛋白-6而增加M2型巨噬细胞特异性分子精氨酸酶-1、IL-10和CD206 mRNA的表达,使巨噬细胞表型从M1(炎症型)转为M2(抗炎型),在神经病理性疼痛的治疗中发挥镇痛作用[23]。Okutani等[24]发现,IL-4可通过降低CC趋化因子配体-3和CC趋化因子受体-5的水平而抑制p38丝裂原活化蛋白激酶活化以及神经元与胶质细胞之间的信息传递,进而抑制脊髓胶质细胞活化,在神经病理性疼痛中发挥镇痛作用。本研究结果显示,CNE组和CAE组小鼠IL-4水平高于CS组,差异有统计学意义,尤其是CAE组虽与CNE组进行一致的低强度运动,但由于运动前30 min注射IL-4Ra阻断运动经IL-4发挥抗炎镇痛作用,因此在行为学测试和抗(促)炎因子水平方面均与CS组结果无异且劣于CNE组,提示神经病理性疼痛小鼠进行低强度运动所发挥的镇痛作用与IL-4有关,但并不能排除低强度运动可能通过其他机制抑制TNF-α、IL-1β、BDNF、β-NGF等细胞因子而发挥镇痛作用,该结果有待进一步研究证实。
综上所述,坐骨神经缩窄性损伤后,局部损伤部位和脊髓均会出现促炎因子释放增加和抗炎因子释放减少而导致机体痛觉过敏,而规律低强度运动2周后可由IL-4介导抗炎因子释放增加和促炎因子释放减少而导致机体痛阈上升,从而发挥镇痛作用。
  • 宁波市自然科学基金(202003N4246)
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2021年第46卷第5期
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doi: 10.11855/j.issn.0577-7402.2021.05.06
  • 接收时间:2021-01-21
  • 首发时间:2025-12-24
  • 出版时间:2021-05-28
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  • 收稿日期:2021-01-21
  • 修回日期:2021-03-08
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Natural Science Foundation of Ningbo City(202003N4246)
宁波市自然科学基金(202003N4246)
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    1浙江省宁波市医疗中心李惠利医院麻醉科,浙江 宁波 315000
    2浙江省宁波市医疗中心李惠利医院手术室,浙江 宁波 315000

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2种不同金属材料的力学参数

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种数
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species
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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