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Article(id=1297570994181402816, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20260047, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1768579200000, receivedDateStr=2026-01-17, revisedDate=null, revisedDateStr=null, acceptedDate=1773590400000, acceptedDateStr=2026-03-16, onlineDate=1787294632715, onlineDateStr=2026-08-21, pubDate=1785772800000, pubDateStr=2026-08-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1787294632715, onlineIssueDateStr=2026-08-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1787294632715, creator=13701087609, updateTime=1787294632715, updator=13701087609, issue=Issue{id=1297570992835023717, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='8', pageStart='3681', pageEnd='4288', issueExtLink='null', onlineDate='null', pubDate='1785772800000', pubDateStr='2026-08-04', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1787294632395, creator='13701087609', updateTime=1787294931551, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1297572247670124783, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1297572247670124784, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=3902, endPage=3912, ext={EN=ArticleExt(id=1297570994386923713, articleId=1297570994181402816, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Efficacy and mechanism of Lacticaseibacillus paracasei CCFM1538 in relieving inflammatory pain in mice, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

[Objective] To investigate the alleviating effects and mechanisms of Lacticaseibacillus paracasei CCFM1538 on inflammatory pain in mice. [Methods] A mouse model of inflammatory pain was induced by complete Freund’s adjuvant, and blank, model, dexamethasone, culture medium, fermentation supernatant, and high-dose/low-dose bacterial groups were set up. The ability of CCFM1538 to alleviate inflammatory pain was evaluated based on paw thickness, thermal/mechanical withdrawal threshold, HE staining of paw skin, inflammatory cytokines, glial cell immunofluorescence, and oxidative stress indicators. [Results] The fermentation supernatant of CCFM1538 significantly alleviated mouse paw swelling, increased the mechanical withdrawal threshold after modeling, reduced paw inflammatory cell infiltration, and decreased the expression of pro-inflammatory cytokines (IL-6 and CCL2). High-dose CCFM1538 cells significantly reduced the expression of pro-inflammatory cytokines (IL-1β and CCL2), decreased serum malondialdehyde levels, and inhibited abnormal activation of spinal cord glial cells. In addition, both significantly increased the thermal withdrawal threshold and serum superoxide dismutase activity in mice after modeling. [Conclusion] The fermentation supernatant and high-dose cells of CCFM1538 can alleviate inflammatory pain by reducing inflammation, alleviating oxidative stress, and regulating spinal glial cell activation. This study provides experimental evidence for the development of related functional foods.

, authors=Huai’en JIANG, Xin TANG, Qiuxiang ZHANG, Jianxin ZHAO, Bingyong MAO, Shumao CUI, authorsList=Huai’en JIANG, Xin TANG, Qiuxiang ZHANG, Jianxin ZHAO, Bingyong MAO, Shumao CUI, authorCompany=null, correspAuthors=Bingyong MAO, authorNote=null, correspAuthorsNote=
E-mail:
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【目的】 探究副干酪乳酪杆菌CCFM1538对小鼠炎性疼痛的缓解效果及作用机制。 【方法】 采用完全弗氏佐剂诱导小鼠炎性疼痛模型，设空白组、模型组、地塞米松组、培养基组、发酵上清组、高/低剂量菌体组。通过测定足部厚度、热/机械缩足阈值、足部皮肤HE染色及炎性细胞因子、神经胶质细胞免疫荧光、氧化应激等指标评价CCFM1538缓解炎性疼痛的能力。 【结果】 CCFM1538发酵上清可显著缓解小鼠造模足部肿胀，提高造模后小鼠机械缩足阈值，减轻足部炎性细胞浸润，并降低促炎因子(IL-6、CCL2)表达；CCFM1538高剂量菌体可显著降低促炎因子(IL-1β、CCL2)表达，减少血清丙二醛含量，并抑制脊髓神经胶质细胞异常活化。此外，二者均可显著提高造模后小鼠热缩足阈值及血清超氧化物歧化酶活性。 【结论】 CCFM1538发酵上清与高剂量菌体可通过减轻炎症、缓解氧化应激及调节脊髓胶质细胞活化等途径缓解炎性疼痛，为相关功能性食品的开发提供了实验依据。

, authors=蒋淮恩, 唐鑫, 张秋香, 赵建新, 毛丙永, 崔树茂, authorsList=蒋淮恩, 唐鑫, 张秋香, 赵建新, 毛丙永, 崔树茂, authorCompany=null, correspAuthors=毛丙永, authorNote=

作者贡献声明

蒋淮恩：实验操作、数据收集与分析、初稿撰写、论文修改；唐鑫、张秋香：实验指导、审阅；赵建新：提供资源、论文修改；毛丙永：实验指导、审阅、论文修改；崔树茂：研究构思和设计、审阅、论文修改。

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Brain, 2024, 147(9): 3157-3170., articleTitle=Nav1.8 in small dorsal root ganglion neurons contributes to vincristine-induced mechanical allodynia, refAbstract=null), Reference(id=1297571009989734689, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297570994181402816, doi=null, pmid=null, pmcid=null, year=2015, volume=6, issue=null, pageStart=10049, pageEnd=null, url=null, language=null, rfNumber=[24], rfOrder=24, authorNames=Leipold E, Hanson-Kahn A, Frick M, Gong P, Bernstein JA, Voigt M, Katona I, Goral RO, Altmüller J, Nürnberg P, Weis J, Hübner CA, Heinemann SH, Kurth I, journalName=Nature Communications, refType=null, unstructuredReference=Leipold E, Hanson-Kahn A, Frick M, Gong P, Bernstein JA, Voigt M, Katona I, Goral RO, Altmüller J, Nürnberg P, Weis J, Hübner CA, Heinemann SH, Kurth I. Cold-aggravated pain in humans caused by a hyperactive NaV1.9 channel mutant[J]. 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Scale bar: 40 μm. *P<0.05, **P<0.01, ***P<0.001 vs. the Model group., figureFileSmall=b9ejWbOr5Jo+6G+WHZB/gg==, figureFileBig=Ltc0HjFGaIPX15fd5rkIbQ==, tableContent=null), ArticleFig(id=1297571005069816069, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297570994181402816, language=CN, label=图6, caption=小鼠脊髓神经胶质细胞免疫荧光, figureFileSmall=b9ejWbOr5Jo+6G+WHZB/gg==, figureFileBig=Ltc0HjFGaIPX15fd5rkIbQ==, tableContent=null), ArticleFig(id=1297571005145313542, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297570994181402816, language=EN, label=Table 1, caption=

Primer sequences used in this study

, figureFileSmall=null, figureFileBig=null, tableContent=
GeneForward primer (5′→3′)Reverse primer (5′→3′)
IL-6TAGTCCTTCCTACCCCAATTTCCTTGGTCCTTAGCCACTCCTTC
IL-1βTTCAGGCAGGCAGTATCACTCGAAGGTCCACGGGAAAGACAC
CCL2TTTGGGTCTCCTTGTTCTCCTGTGAAATTAGGGCAGCAGGACAT
GAPDHAGAAGGTGGTGAAGCAGGCATCCGAAGGTGGAAGAGTGGGAGTTG
), ArticleFig(id=1297571005245976839, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297570994181402816, language=CN, label=表1, caption=

本研究所用引物

, figureFileSmall=null, figureFileBig=null, tableContent=
GeneForward primer (5′→3′)Reverse primer (5′→3′)
IL-6TAGTCCTTCCTACCCCAATTTCCTTGGTCCTTAGCCACTCCTTC
IL-1βTTCAGGCAGGCAGTATCACTCGAAGGTCCACGGGAAAGACAC
CCL2TTTGGGTCTCCTTGTTCTCCTGTGAAATTAGGGCAGCAGGACAT
GAPDHAGAAGGTGGTGAAGCAGGCATCCGAAGGTGGAAGAGTGGGAGTTG
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副干酪乳酪杆菌CCFM1538缓解小鼠炎性疼痛的功效及机制
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蒋淮恩 , 唐鑫 , 张秋香 , 赵建新 , 毛丙永 , 崔树茂
微生物学报 | 研究报告 2026,66(8): 3902-3912
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微生物学报 |研究报告 2026 , 66 (8) : 3902 -3912
副干酪乳酪杆菌CCFM1538缓解小鼠炎性疼痛的功效及机制
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蒋淮恩, 唐鑫, 张秋香, 赵建新, 毛丙永 , 崔树茂
作者信息
  • 江南大学 食品学院，江苏 无锡
通讯作者:
毛丙永
作者简介:

作者贡献声明

蒋淮恩：实验操作、数据收集与分析、初稿撰写、论文修改；唐鑫、张秋香：实验指导、审阅；赵建新：提供资源、论文修改；毛丙永：实验指导、审阅、论文修改；崔树茂：研究构思和设计、审阅、论文修改。

Efficacy and mechanism of Lacticaseibacillus paracasei CCFM1538 in relieving inflammatory pain in mice
Huai’en JIANG, Xin TANG, Qiuxiang ZHANG, Jianxin ZHAO, Bingyong MAO , Shumao CUI
Affiliations
  • School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China
    • Corresponding Author:
    E-mail:
出版时间: 2026-08-04 doi: 10.13343/j.cnki.wsxb.20260047
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【目的】 探究副干酪乳酪杆菌CCFM1538对小鼠炎性疼痛的缓解效果及作用机制。 【方法】 采用完全弗氏佐剂诱导小鼠炎性疼痛模型，设空白组、模型组、地塞米松组、培养基组、发酵上清组、高/低剂量菌体组。通过测定足部厚度、热/机械缩足阈值、足部皮肤HE染色及炎性细胞因子、神经胶质细胞免疫荧光、氧化应激等指标评价CCFM1538缓解炎性疼痛的能力。 【结果】 CCFM1538发酵上清可显著缓解小鼠造模足部肿胀，提高造模后小鼠机械缩足阈值，减轻足部炎性细胞浸润，并降低促炎因子(IL-6、CCL2)表达；CCFM1538高剂量菌体可显著降低促炎因子(IL-1β、CCL2)表达，减少血清丙二醛含量，并抑制脊髓神经胶质细胞异常活化。此外，二者均可显著提高造模后小鼠热缩足阈值及血清超氧化物歧化酶活性。 【结论】 CCFM1538发酵上清与高剂量菌体可通过减轻炎症、缓解氧化应激及调节脊髓胶质细胞活化等途径缓解炎性疼痛，为相关功能性食品的开发提供了实验依据。

后生元  /  副干酪乳酪杆菌  /  完全弗氏佐剂  /  炎性疼痛  /  胶质细胞

[Objective] To investigate the alleviating effects and mechanisms of Lacticaseibacillus paracasei CCFM1538 on inflammatory pain in mice. [Methods] A mouse model of inflammatory pain was induced by complete Freund’s adjuvant, and blank, model, dexamethasone, culture medium, fermentation supernatant, and high-dose/low-dose bacterial groups were set up. The ability of CCFM1538 to alleviate inflammatory pain was evaluated based on paw thickness, thermal/mechanical withdrawal threshold, HE staining of paw skin, inflammatory cytokines, glial cell immunofluorescence, and oxidative stress indicators. [Results] The fermentation supernatant of CCFM1538 significantly alleviated mouse paw swelling, increased the mechanical withdrawal threshold after modeling, reduced paw inflammatory cell infiltration, and decreased the expression of pro-inflammatory cytokines (IL-6 and CCL2). High-dose CCFM1538 cells significantly reduced the expression of pro-inflammatory cytokines (IL-1β and CCL2), decreased serum malondialdehyde levels, and inhibited abnormal activation of spinal cord glial cells. In addition, both significantly increased the thermal withdrawal threshold and serum superoxide dismutase activity in mice after modeling. [Conclusion] The fermentation supernatant and high-dose cells of CCFM1538 can alleviate inflammatory pain by reducing inflammation, alleviating oxidative stress, and regulating spinal glial cell activation. This study provides experimental evidence for the development of related functional foods.

postbiotics  /  Lacticaseibacillus paracasei  /  complete Freund’s adjuvant  /  inflammatory pain  /  glial cells
蒋淮恩, 唐鑫, 张秋香, 赵建新, 毛丙永, 崔树茂. 副干酪乳酪杆菌CCFM1538缓解小鼠炎性疼痛的功效及机制. 微生物学报, 2026 , 66 (8) : 3902 -3912 . DOI: 10.13343/j.cnki.wsxb.20260047
Huai’en JIANG, Xin TANG, Qiuxiang ZHANG, Jianxin ZHAO, Bingyong MAO, Shumao CUI. Efficacy and mechanism of Lacticaseibacillus paracasei CCFM1538 in relieving inflammatory pain in mice[J]. Acta Microbiologica Sinica, 2026 , 66 (8) : 3902 -3912 . DOI: 10.13343/j.cnki.wsxb.20260047
正文
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炎性疼痛是关节炎、肠炎等多种炎症性疾病的共同表现，其发生与局部及全身性炎症反应密切相关[1]。炎性疼痛波及全球数百万人并严重影响患者生活质量，造成巨大的医疗负担和社会经济压力[2]。目前临床上常用的缓解炎性疼痛的药物主要有阿片类药物、非甾体抗炎药等，这些药物通过抑制炎症或阻断痛觉信号传导等途径缓解疼痛。然而长期使用常伴随着成瘾性、胃肠道不适等副作用，且在部分慢性疼痛中疗效受限[3-4]。因此，开发安全、有效、副作用小的缓解疼痛策略具有重要意义。
近年来，肠道菌群及其代谢物受到广泛关注，其与宿主健康的关系日益明晰。研究显示，特定益生菌及其代谢物可通过调节免疫[5]、缓解炎症[6]等方式影响疼痛感知。随着益生菌研究的深入，已发现益生菌的益生功效不局限于活菌及其代谢物，其灭活菌体成分同样具有重要的益生作用。后生元(postbiotics)被定义为“对宿主健康有益的无生命微生物和/或其成分的制剂”，作为非活菌制剂，包括灭活菌体、细胞碎片、可包含或不包含其代谢产物等组分[7]。与活菌制剂相比，后生元不仅具备良好的加工稳定性和储存便利性，还能避免活菌可能带来的生物安全风险，在功能食品和特医食品中显示出广阔前景[8]。此外，相较于活菌，后生元具备独特的生物功能，有研究显示热灭活的嗜黏蛋白阿克曼氏菌(Akkermansia muciniphila)可显著降低肥胖人群体重、胰岛素水平，而补充活菌的总体益处不显著[9]。然而，目前关于后生元在炎性疼痛中的作用仍缺乏系统研究，其是否通过调节神经胶质细胞活性、氧化应激等途径发挥镇痛效应尚不明确。
本研究选取实验室自主分离保存的副干酪乳酪杆菌(Lacticaseibacillus paracasei) CCFM1538，以其发酵上清和灭活菌体作为研究对象，通过完全弗氏佐剂(complete Freund’s adjuvant, CFA)诱导的小鼠足底炎性疼痛模型，综合评价其镇痛作用，并进一步从足部肿胀、组织炎症、促炎细胞因子表达、脊髓胶质细胞活化及系统氧化应激等维度探讨其潜在机制，以期为缓解疼痛相关功能食品的开发提供理论支持。
1 材料与方法
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1.1 菌株
副干酪乳酪杆菌CCFM1538 (分离于藏族人群粪便)，保藏于江南大学食品微生物保藏中心。
1.2 主要试剂和仪器
地塞米松，MedChemexpress生物科技公司；组织总RNA提取试剂、通用型逆转录酶、qPCR反应体系，南京诺唯赞生物科技股份有限公司；多聚甲醛，武汉赛维尔生物科技有限公司；CFA，默克公司；超氧化物歧化酶(superoxide dismutase, SOD)检测试剂盒、丙二醛(malondialdehyde, MDA)检测试剂盒，南京建成生物工程研究所有限公司。
落地式离心机、多功能酶标仪，ThermoFisher Scientific公司；真空冷冻干燥机，泰式达公司；实时荧光定量PCR仪，Bio-Rad Laboratories公司；数字切片扫描仪，3D HISTECH有限公司；智能热板仪，上海欣软信息科技有限公司；Von Frey纤维丝，DanMic公司。
1.3 CCFM1538后生元制备
将CCFM1538接种于MRS培养基中活化3代，获得种子液后按2%接种量接入MRS液体培养基中，37 ℃培养12-18 h。发酵液经4 ℃、8 000×g离心20 min，上清过0.22 μm滤膜，pH值为3.8±0.1。正常MRS液体培养基过0.22 μm滤膜后调节pH值为3.8±0.1，于-20 ℃保存备用。
菌泥经生理盐水清洗2-3次后，用生理盐水重悬并进行巴氏灭菌(65 ℃、30 min)，之后进行平板菌落计数，确保无活菌或少于103 CFU/g后进行冷冻干燥，冻干菌粉使用生理盐水复溶后-20 ℃保存备用。菌泥和发酵液灌胃样品在动物灌胃前1周内制备完成。
1.4 动物实验
1.4.1 动物实验设计
56只无特定病原体级(specific pathogen free, SPF)雄性C57BL/6J小鼠(8周龄，体重20-22 g)购自北京斯贝福生物技术有限公司。小鼠饲养于标准环境，自由进食和饮水。本研究所有动物实验获得江南大学动物伦理委员会批准，编号为JN.No20250615c1120726。
适应性饲养7 d后，小鼠随机分为7组(n=8)：空白组(Control)、模型组(Model)、地塞米松组(Dexamethasone)、培养基组(Medium)、CCFM1538发酵上清组(CCFM1538-FS)、CCFM1538高剂量菌体组(CCFM1538-H)、CCFM1538低剂量菌体组(CCFM1538-L)。除空白组于左后足底皮下注射20 μL无菌生理盐水外，其余组别注射20 μL CFA建立炎性疼痛模型[10]。造模前1天和造模后1天进行热缩足阈值实验和机械缩足阈值实验，以评估造模是否成功。
造模后第1天开始干预，每日灌胃，持续14 d。空白组、模型组灌胃200 μL生理盐水；培养基组灌胃200 μL液体培养基；CCFM1538发酵上清组灌胃200 μL发酵上清液；CCFM1538高、低剂量菌体组分别每日灌胃200 μL含1×1010 CFU与2×109 CFU灭活菌体悬液；阳性药物组腹腔注射地塞米松(1 mg/kg，溶于200 μL生理盐水)[11]，阳性药物组只用于验证模型与测定体系敏感性，而非与试验组做严格等剂量等途径比较。
1.4.2 足部厚度测量
使用游标卡尺测量小鼠造模侧足部厚度。
1.4.3 行为学测试
于造模前1天(基线)及造模后第1、3、5、14天进行。
采用(55±0.5) ℃智能热板仪测试热缩足阈值。将小鼠置于热板，记录其出现舔足、抬足或跳跃的潜伏期，截止时间设为30 s[12]。
采用Von Frey纤维丝法测试机械缩足阈值。将小鼠置于网状底座观察箱，适应后，使用一系列不同克重的纤维丝垂直刺激左后足底中部，持续3-5 s。以“up-and-down法”确定50%机械缩足阈值[13]。
1.4.4 动物样本采集
干预结束后禁食12 h。麻醉后经心脏灌注[14]，取足底皮肤及L4-L6脊髓组织用于组织学与免疫荧光分析；未进行心脏灌注的小鼠通过眼眶静脉丛采血，收集血液样本静置1 h后，以4 ℃、3 000×g离心15 min，收集上层血清并保存于-80 ℃冰箱以备后续分析。颈椎脱臼处死小鼠，收集小鼠造模部位足底皮肤并保存于-80 ℃冰箱。
1.4.5 组织病理学分析
皮肤组织经4%多聚甲醛固定48 h后，石蜡包埋、切片，进行苏木精-伊红染色(hematoxylin and eosin staining, HE staining)。使用数字切片扫描仪对染色后的皮肤切片进行扫描，观察和评估皮肤组织的病理变化。
1.4.6 实时荧光定量逆转录PCR分析
采用试剂盒提取小鼠皮肤组织样本中的总RNA，并逆转录为cDNA。以GAPDH基因作为内参，检测炎症相关的目标基因白细胞介素1β (interleukin-1β, IL-1β)、白细胞介素-6 (interleukin-6, IL-6)和趋化因子2 (C-C motif ligand 2, CCL2) mRNA相对表达水平。PCR反应体系(10 μL)：2×ChamQ Universal SYBR qPCR Master Mix 5 µL，上、下游引物(10 µmol/L)各0.25 µL，cDNA模板1 µL，ddH2O 3.5 µL。PCR反应条件：95 ℃预变性30 s；95 ℃变性10 s，退火30 s，60 ℃延伸30 s，共40个循环。通过2-ΔΔCt法定量分析目标基因mRNA的相对表达水平，具体的引物序列信息列于表1。
1.4.7 血清SOD、MDA检测
按照SOD和MDA检测试剂盒说明书操作，测定血清中SOD活性和MDA含量。
1.4.8 免疫荧光染色分析
取灌注固定的脊髓组织制备冷冻切片，切片经抗原修复、封闭后，分别与IBA-1 (小胶质细胞标志物)及GFAP (星形胶质细胞标志物)一抗4 ℃孵育过夜，再与相应荧光二抗室温孵育。封片后使用数字切片扫描仪采集图像，并用ImageJ软件定量分析目标蛋白的平均荧光强度。
1.5 统计学方法
使用GraphPad Prism 10.0软件进行统计分析。行为学与足部厚度数据使用双因素方差分析(two-way ANOVA)进行处理，随后进行Bonferroni检验校正多重比较。其他数据使用单因素方差分析(one-way ANOVA)进行处理，随后进行Dunnett检验校正多重比较。
2 结果与分析
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2.1 CCFM1538对小鼠造模侧足部肿胀的影响
小鼠足底注射CFA会引起严重的炎症反应，其典型外在表现为注射后足部红肿[15]。如图1所示，与空白组相比，其余组别小鼠在注射CFA后第1天均出现显著的足部肿胀。干预14 d后，与模型组相比，阳性药物地塞米松组小鼠足部厚度显著降低(P<0.05)，培养基组与模型组相似，表明培养基本身无消肿作用；在CCFM1538干预组中，发酵上清显著降低小鼠足部厚度(P<0.05)，而高/低剂量菌体对足部肿胀改善不显著。上述结果表明，CCFM1538发酵上清液能够有效缓解CFA诱导的足部肿胀。
2.2 CCFM1538对小鼠炎性疼痛行为学指标的影响
为进一步研究CCFM1538对CFA诱导的小鼠炎性疼痛的缓解作用，本研究通过2种典型的疼痛行为评估其疼痛缓解效果。如图2所示，与空白组相比，小鼠在CFA造模后第1天的热缩足与机械缩足阈值均显著降低，表明小鼠炎性疼痛模型诱导成功。干预后，与模型组相比，阳性药物地塞米松组小鼠热缩足阈值在第3天显著恢复(P<0.001)、机械缩足阈值在第5天显著恢复(P<0.05)，证实其具有明确的镇痛效果；培养基组与模型组相似，表明培养基本身对疼痛无缓解作用；在CCFM1538干预组中，发酵上清和高剂量菌体均在干预第3天显著提升热缩足阈值(P<0.01、P<0.001)；发酵上清在干预第14天还能显著提高机械缩足阈值(P<0.05)，而高/低剂量菌体对机械痛阈改善不显著。以上数据表明，CCFM1538发酵上清在缓解机械性痛觉过敏方面作用更为突出，而高剂量菌体与发酵上清在热痛敏缓解中均表现显著。
2.3 CCFM1538对小鼠足部组织炎症的影响
为直接观察CFA注射后小鼠足部皮肤及皮下组织的炎性细胞浸润、肿胀等病理变化，本研究对注射侧足部皮肤进行HE染色[16]。如图3所示，与空白组相比，模型组小鼠皮肤出现严重的炎症性病理改变，具体表现为真皮层出现大量炎性细胞浸润并伴有肉芽肿形成，真皮层和皮下组织间隙增宽、结构疏松，局部血管扩张、充血。阳性药物地塞米松干预后，炎性细胞浸润明显减轻，真皮层与皮下组织结构基本恢复正常。培养基组的病理表现与模型组相似，表明培养基本身对炎症无改善作用。CCFM1538发酵上清液干预后炎症改善最为明显，炎性细胞浸润显著减少，组织结构恢复接近正常；CCFM1538高、低剂量菌体干预后虽有一定程度的改善，但仍可见较明显的炎性细胞浸润。上述组织学结果表明，CCFM1538发酵上清液能够有效缓解CFA诱导的皮肤炎症损伤。
2.4 CCFM1538对小鼠皮肤促炎细胞因子表达的影响
研究表明，炎症因子和趋化因子会导致持续的慢性疼痛，因此细胞因子水平可以间接反映疼痛水平[17]。如图4所示，与空白组相比，模型组小鼠皮肤组织中促炎细胞因子IL-1β、IL-6和CCL2的mRNA水平均显著提升(P<0.001、P<0.001、P<0.001)。阳性药物地塞米松干预可显著降低上述所有细胞因子的mRNA水平(P<0.05、P<0.001、P<0.001)，而培养基组各细胞因子水平与模型组相比均无显著差异，表明培养基本身不具备抗炎作用；发酵上清可显著降低IL‑6与CCL2表达(P<0.01、P<0.01)，而高剂量菌体主要下调IL‑1β与CCL2表达(P<0.01、P<0.01)。该结果表明，CCFM1538后生元可通过选择性抑制不同促炎因子的表达发挥抗炎作用，且发酵上清与菌体的作用靶点存在差异。
2.5 CCFM1538对小鼠血清氧化应激水平的影响
氧化应激是炎性疼痛的关键机制之一，炎症刺激导致活性氧过量产生，加剧炎症反应和痛觉过敏[18]。如图5所示，与空白组(MDA 7.25 nmol/mL、SOD 47.98 U/mL)相比，模型组小鼠出现显著的氧化应激状态(MDA 9.25 nmol/mL，P<0.01；SOD 36.75 U/mL，P<0.05)，说明CFA诱导的炎性疼痛伴随着氧化应激反应。阳性药物地塞米松干预后MDA含量和SOD活性恢复至正常水平(MDA 7.75 nmol/mL，P<0.05；SOD 47.77 U/mL，P<0.05)，表明药物处理可以缓解炎性疼痛模型小鼠氧化应激状态；培养基组MDA含量有所降低(9 nmol/mL)、SOD活性(44.74 U/mL)高于模型组，但无统计学意义。CCFM1538发酵上清与高剂量菌体干预后，SOD活性均显著升高(50.67 U/mL，P<0.05；50.91 U/mL，P<0.01)，并且高剂量菌体干预还显著降低MDA含量(7.75 nmol/mL, P<0.05)。这表明二者能有效缓解CFA引起的系统氧化应激状态，增强机体抗氧化能力，这可能与其抑制炎症、调节免疫微环境有关。
2.6 CCFM1538对小鼠脊髓胶质细胞活性的影响
当疼痛信号由初级背根神经节传入神经传递至脊髓背角时，传入神经释放谷氨酸、ATP和趋化因子，激活脊髓背角二阶神经元，将疼痛信号传递至大脑。同时，ATP和趋化因子会激活脊髓小胶质细胞和星形胶质细胞，活化的神经胶质细胞释放促炎细胞因子，作用于传入神经元和二阶神经元，增强其兴奋性与神经递质传递，并介导中枢疼痛敏化[19-20]。神经胶质细胞活化是中枢疼痛敏化的核心环节，是评价疼痛状态的重要表征指标。
如图6所示，模型组小鼠脊髓组织中出现明显的小胶质细胞与星形胶质细胞异常活化。与空白组相比，模型组小胶质细胞胞体肥大、呈阿米巴样；星形胶质细胞胞体也肥大，突起增粗，说明炎性疼痛的产生伴随着显著的神经胶质细胞异常活化反应。阳性药物地塞米松干预可有效抑制星形胶质细胞(P<0.01)、小胶质细胞(P<0.05)的活化状态，使其恢复至正常水平；培养基组与模型组相似，无改善作用。
高剂量菌体能同时抑制2种胶质细胞的异常活化(P<0.01、P<0.05)，而发酵上清在此模型中未表现出显著抑制作用。这一结果提示，灭活菌体可能通过调节神经胶质细胞介导的中枢敏化途径缓解疼痛，而发酵上清的作用更侧重于外周炎症环节。
3 讨论与结论
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本研究通过体内实验证实副干酪乳酪杆菌CCFM1538能有效缓解CFA诱导的小鼠炎性疼痛，其中发酵上清主要通过下调外周组织IL-6和CCL2的表达、减轻局部炎性浸润发挥作用；高剂量灭活菌体则侧重于抑制IL-1β、CCL2的表达，提高抗氧化能力并调节脊髓小胶质细胞与星形胶质细胞的活化状态，提示其可能从中枢层面干预疼痛信号传递。这些发现提示副干酪乳酪杆菌CCFM1538可通过外周和中枢层面影响疼痛，但需要进一步的因果验证以明确其具体作用靶点。
本研究为缓解疼痛相关功能性食品的开发提供了理论依据与物质基础，未来研究可在多个层面深入探讨副干酪乳酪杆菌CCFM1538缓解炎性疼痛的作用机制。在物质层面，可以将菌体、发酵上清细分为胞外囊泡、肽聚糖等组分，并在细胞、动物、类器官等模型中进一步发掘其功效成分；在机制层面，本研究提供了初步的线索，如炎症因子变化与神经胶质细胞活化状态改变等；要明确其机制，未来研究需进行因果验证，包括：采用胶质细胞抑制剂/拮抗剂验证神经胶质细胞在镇痛效应中的作用；检测瞬时受体电位离子通道TRPV1[21]、TRPA1[22]及电压门控钠通道NaV1.8[23]、NaV1.9[24]的磷酸化状态，并以此发掘调控离子通路的上游信号通路，明确通路之间的相互作用，进一步阐明副干酪乳酪杆菌CCFM1538镇痛的作用机制；在模型层面，本研究在CFA诱导的小鼠炎性疼痛模型中验证了其功效，可进一步在其他疼痛模型，如坐骨神经慢性挤压模型、紫杉醇神经病理性疼痛模型、糖尿病神经病理性疼痛模型中验证其功效。

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doi: 10.13343/j.cnki.wsxb.20260047
  • 接收时间：2026-01-17
  • 首发时间：2026-08-21
  • 出版时间：2026-08-04
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  • 收稿日期：2026-01-17
  • 录用日期：2026-03-16
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    江南大学 食品学院，江苏 无锡

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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
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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