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Article(id=1297571047004463863, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20260161, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1772121600000, receivedDateStr=2026-02-27, revisedDate=null, revisedDateStr=null, acceptedDate=1776268800000, acceptedDateStr=2026-04-16, onlineDate=1787294645308, onlineDateStr=2026-08-21, pubDate=1785772800000, pubDateStr=2026-08-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1787294645308, onlineIssueDateStr=2026-08-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1787294645308, creator=13701087609, updateTime=1787294645308, 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=3953, endPage=3964, ext={EN=ArticleExt(id=1297571047222567672, articleId=1297571047004463863, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Inactivated cells of Bifidobacterium animalis CCFM1155 modulate the Nrf2 signaling pathway to alleviate skin aging in mice, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

[Objective] To investigate the efficacy of postbiotics prepared with Bifidobacterium animalis CCFM1155, a strain previously identified for its ability to enhance nuclear factor erythroid 2-related factor 2 (Nrf2) secretion in human immortalized epidermal cells, in alleviating skin aging. [Methods] A mouse model of skin aging was established by daily subcutaneous injection of D-galactose (500 mg/kg) for eight weeks (n=6 per group). The model mice were then orally administered either heat-inactivated cells (1155B) or concentrated fermentation supernatant (1155T) of CCFM1155 cells at a dose equivalent to 5×109 CFU of live bacteria. [Results] The 1155B was the primary postbiotic component exerting anti-skin aging effects, whereas the 1155T group showed no significant effect. Compared with the model group, the 1155B treatment ameliorated skin aging phenotypes by improving the skin tissue structure, increasing the epidermal thickness (P<0.05) and dermal collagen fiber density (P<0.01), and decreasing the expression of the senescence marker protein p21 and the activity of senescence-associated β-galactosidase (P<0.001). In addition, the 1155B treatment enhanced the antioxidant defense system of the skin by elevating the activities of total superoxide dismutase (P<0.05) and catalase (P<0.01), as well as the content of reduced glutathione (P<0.01), while lowering the malondialdehyde level (P<0.01). Mechanism studies revealed that the 1155B upregulated both the mRNA (P<0.05) and protein (P<0.001) levels of Nrf2 in the skin. Consequently, it promoted the transcription of key downstream antioxidant molecules, including heme oxygenase-1, peroxiredoxin, thioredoxin 1, and the glutamate-cysteine ligase catalytic subunit, thereby activating the downstream signaling pathway of Nrf2. [Conclusion] The inactivated cells of B. animalis CCFM1155 alleviates skin aging by activating the Nrf2 signaling pathway. These findings offer a novel perspective on postbiotic regulation of the Nrf2 pathway and lay a theoretical foundation for developing safe and effective anti-skin aging products.

, authors=Haobo CUI, Xin TANG, Bingyong MAO, Qiuxiang ZHANG, Qixiao ZHAI, Jianxin ZHAO, Wei CHEN, Shumao CUI, authorsList=Haobo CUI, Xin TANG, Bingyong MAO, Qiuxiang ZHANG, Qixiao ZHAI, Jianxin ZHAO, Wei CHEN, Shumao CUI, authorCompany=null, correspAuthors=Xin TANG, authorNote=null, correspAuthorsNote=
E-mail:
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【目的】 探究能够提高人永生化角质形成细胞(human immortalized epidermal cells, HaCaT)核转录因子E2相关因子2 (nuclear factor erythroid 2-related factor 2, Nrf2)表达量的动物双歧杆菌CCFM1155，口服其制备的后生元调控皮肤Nrf2信号通路并缓解皮肤衰老。 【方法】 通过每日皮下注射500 mg/kg D-半乳糖诱导皮肤衰老小鼠模型(n=6)，并分别灌胃5×109 CFU等量的动物双歧杆菌CCFM1155灭活菌体和浓缩上清发酵液，8周后检测小鼠皮肤衰老状况。 【结果】 动物双歧杆菌CCFM1155后生元中发挥主要抗皮肤衰老作用的成分为灭活菌体，而浓缩上清发酵液组未表现出明显改善效果。与模型组相比，口服CCFM1155灭活菌体组(1155B)能显著改善衰老小鼠皮肤组织结构，显著增加表皮厚度(P<0.05)与真皮胶原纤维密度(P<0.01)，降低衰老标志蛋白p21的表达及β-半乳糖苷酶活性(P<0.001)，有效缓解皮肤衰老表型。此外，1155B组还显著提高了皮肤中总超氧化物歧化酶(P<0.05)、过氧化氢酶活性(P<0.01)及还原型谷胱甘肽含量(P<0.01)，降低丙二醛水平(P<0.01)，增强整体抗氧化能力。进一步机制研究表明，灭活菌体可上调Nrf2的mRNA (P<0.05)和蛋白表达(P<0.001)，并促进其下游抗氧化基因血红素加氧酶-1 (heme oxygenase-1, HO-1)、过氧化物还原酶1 (peroxiredoxin 1, Prdx1)、硫氧还蛋白1 (thioredoxin 1, TXN1)和谷氨酸半胱氨酸连接酶催化亚基(glutamate-cysteine ligase catalytic subunit, GCLC)的转录，从而激活Nrf2下游信号通路。 【结论】 本研究初步证实动物双歧杆菌CCFM1155灭活菌体可通过激活Nrf2信号通路缓解皮肤衰老。该研究为后生元调控Nrf2通路提供了新视角，为开发安全有效的抗皮肤衰老产品奠定了理论基础。

, authors=崔皓博, 唐鑫, 毛丙永, 张秋香, 翟齐啸, 赵建新, 陈卫, 崔树茂, authorsList=崔皓博, 唐鑫, 毛丙永, 张秋香, 翟齐啸, 赵建新, 陈卫, 崔树茂, authorCompany=null, correspAuthors=唐鑫, authorNote=

作者贡献声明

崔皓博：实验规划、数据整理与分析、文章撰写；唐鑫：实验指导、论文修改；毛丙永：文章构思；张秋香：文章讨论；翟齐啸：提供建议；赵建新：提供资源；陈卫：稿件审阅；崔树茂：监督管理。

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International Journal of Molecular Sciences, 2024, 25(20): 11326., articleTitle=Limosilactobacillus reuteri HY7503 and its cellular proteins alleviate endothelial dysfunction by increasing nitric oxide production and regulating cell adhesion molecule levels, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1297571049495880464, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, xref=null, ext=[AuthorCompanyExt(id=1297571049504269073, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, companyId=1297571049495880464, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China), AuthorCompanyExt(id=1297571049512657682, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, companyId=1297571049495880464, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=江南大学 食品学院，江苏 无锡)])], figs=[ArticleFig(id=1297571052398338886, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 1, caption=HE staining of mouse dorsal skin., figureFileSmall=pnu5ut9kelWKkyoGZmgnOw==, figureFileBig=43D8Ozyh/qY/Vt0niFbgNw==, tableContent=null), ArticleFig(id=1297571052465447751, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图1, caption=小鼠背部皮肤HE染色, figureFileSmall=pnu5ut9kelWKkyoGZmgnOw==, figureFileBig=43D8Ozyh/qY/Vt0niFbgNw==, tableContent=null), ArticleFig(id=1297571052658385736, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 2, caption=Epidermal thickness of mouse dorsal skin. *P<0.05, **P<0.01, ****P<0.000 1 vs. the model group., figureFileSmall=N0zBUtRlcfpgwRtfv+QHIA==, figureFileBig=roTofIHiRw5W1JJoZHcQ5A==, tableContent=null), ArticleFig(id=1297571052725494601, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图2, caption=小鼠背部皮肤表皮厚度, figureFileSmall=N0zBUtRlcfpgwRtfv+QHIA==, figureFileBig=roTofIHiRw5W1JJoZHcQ5A==, tableContent=null), ArticleFig(id=1297571052788409162, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 3, caption=Masson staining of mouse dorsal skin., figureFileSmall=yIHpASjMg5VbE9Ul7n1oGg==, figureFileBig=1J5GxsQrLRcjGW2wz4gVyg==, tableContent=null), ArticleFig(id=1297571052851323723, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图3, caption=小鼠背部皮肤Masson染色, figureFileSmall=yIHpASjMg5VbE9Ul7n1oGg==, figureFileBig=1J5GxsQrLRcjGW2wz4gVyg==, tableContent=null), ArticleFig(id=1297571052926821196, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 4, caption=Relative quantification of collagen in mouse dorsal skin. **P<0.01, ***P<0.001, ****P<0.000 1 vs. the model group., figureFileSmall=h6tcEptYukkJMKPJ7VNuyA==, figureFileBig=0+y+gq45tcrY1sFRg99Rdw==, tableContent=null), ArticleFig(id=1297571053019095885, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图4, caption=小鼠背部皮肤胶原蛋白相对定量, figureFileSmall=h6tcEptYukkJMKPJ7VNuyA==, figureFileBig=0+y+gq45tcrY1sFRg99Rdw==, tableContent=null), ArticleFig(id=1297571053107176270, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 5, caption=Immunohistochemical staining of p21 in mouse skin., figureFileSmall=16i/IAJI/8eyOhNmZXFycA==, figureFileBig=j0BLat1NZqr4e+seS1PVRg==, tableContent=null), ArticleFig(id=1297571053174285135, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图5, caption=小鼠皮肤p21免疫组化染色, figureFileSmall=16i/IAJI/8eyOhNmZXFycA==, figureFileBig=j0BLat1NZqr4e+seS1PVRg==, tableContent=null), ArticleFig(id=1297571053249782608, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 6, caption=β-galactosidase activity in mouse skin. ***P<0.001, ****P<0.000 1 vs. the model group., figureFileSmall=5CJSVdDV5XJFgw6tZEXLRQ==, figureFileBig=/ZZqHt4DsWSo6p9MDSqsRA==, tableContent=null), ArticleFig(id=1297571053321085777, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图6, caption=小鼠皮肤β-半乳糖苷酶活力, figureFileSmall=5CJSVdDV5XJFgw6tZEXLRQ==, figureFileBig=/ZZqHt4DsWSo6p9MDSqsRA==, tableContent=null), ArticleFig(id=1297571053404971858, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 7, caption=Antioxidant capacity indices in mouse skin. A: SOD activity; B: CAT activity; C: GSH level; D: MDA level. *P<0.05, **P<0.01, ***P<0.001, ****P<0.000 1 vs. the model group., figureFileSmall=dPLGDsW6LXljdjmncvFcYQ==, figureFileBig=GOknVmwkJIBp2mmgLl/69w==, tableContent=null), ArticleFig(id=1297571053467886419, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图7, caption=小鼠皮肤抗氧化能力指标, figureFileSmall=dPLGDsW6LXljdjmncvFcYQ==, figureFileBig=GOknVmwkJIBp2mmgLl/69w==, tableContent=null), ArticleFig(id=1297571053539189588, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 8, caption=Relative mRNA expression of Nrf2 and Keap1 in mouse skin. A: Nrf2 mRNA expression; B: Keap1 mRNA expression. *P<0.05, ***P<0.001 vs. the model group., figureFileSmall=pQ8uybbaDO9sMM71FNGPOg==, figureFileBig=7qbhOtPnbqEpSnJAM+gaig==, tableContent=null), ArticleFig(id=1297571053610492757, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图8, caption=小鼠皮肤Nrf2、Keap1的mRNA相对表达, figureFileSmall=pQ8uybbaDO9sMM71FNGPOg==, figureFileBig=7qbhOtPnbqEpSnJAM+gaig==, tableContent=null), ArticleFig(id=1297571053694378838, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 9, caption=Immunohistochemical staining for Nrf2 in mouse skin., figureFileSmall=orSNjzaF7/A7ycW/Y9S/kA==, figureFileBig=Q4xWOJ01xfY7cLVoN+dgNg==, tableContent=null), ArticleFig(id=1297571053774070615, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图9, caption=小鼠皮肤Nrf2免疫组化染色, figureFileSmall=orSNjzaF7/A7ycW/Y9S/kA==, figureFileBig=Q4xWOJ01xfY7cLVoN+dgNg==, tableContent=null), ArticleFig(id=1297571053841179480, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 10, caption=Quantitative analysis of immunohistochemistry. ***P<0.001, ****P<0.000 1 vs. the model group., figureFileSmall=swvd7mq5uX1gDHE1oHh2Fg==, figureFileBig=4Xu4+urD+WkPlnZ/7xRfew==, tableContent=null), ArticleFig(id=1297571053904094041, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图10, caption=免疫组化定量结果, figureFileSmall=swvd7mq5uX1gDHE1oHh2Fg==, figureFileBig=4Xu4+urD+WkPlnZ/7xRfew==, tableContent=null), ArticleFig(id=1297571053962814298, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Figure 11, caption=Relative expression of Nrf2 downstream pathway mRNAs in mouse skin. A: HO-1 mRNA expression; B: Prdx1 mRNA expression; C: TXN1 mRNA expression; D: GCLC mRNA expression. **P<0.01, ***P<0.001, ****P<0.000 1 vs. the model group., figureFileSmall=86yKqfbhNodTZJvLpcfL/w==, figureFileBig=cOBNrM3w8/vpxrcrvuayJw==, tableContent=null), ArticleFig(id=1297571054021534555, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=图11, caption=小鼠皮肤Nrf2下游通路mRNA的相对表达, figureFileSmall=86yKqfbhNodTZJvLpcfL/w==, figureFileBig=cOBNrM3w8/vpxrcrvuayJw==, tableContent=null), ArticleFig(id=1297571054097032028, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=EN, label=Table 1, caption=

Target gene primer sequences

, figureFileSmall=null, figureFileBig=null, tableContent=
Primer namesForward primer sequences (5′→3′)Reverse primer sequences (5′→3′)
Nrf2TCTTGGAGTAAGTCGAGAAGTGTGTTGAAACTGAGCGAAAAAGGC
Keap1TGCCCCTGTGGTCAAAGTGGGTTCGGTTACCGTCCTGC
HO-1AAGCCGAGAATGCTGAGTTCAGCCGTGTAGATATGGTACAAGGA
PRDX1AATGCAAAAATTGGGTATCCTGCCGTGGGACACACAAAAGTAAAGT
TXN1CATGCCGACCTTCCAGTTTTATTTCCTTGTTAGCACCGGAGA
GCLCGGGGTGACGAGGTGGAGTAGTTGGGGTTTGTCCTCTCCC
GAPDHAGGTCGGTGTGAACGGATTTGTGTAGACCATGTAGTTGAGGTCA
), ArticleFig(id=1297571054172529501, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571047004463863, language=CN, label=表1, caption=

目的基因的引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
Primer namesForward primer sequences (5′→3′)Reverse primer sequences (5′→3′)
Nrf2TCTTGGAGTAAGTCGAGAAGTGTGTTGAAACTGAGCGAAAAAGGC
Keap1TGCCCCTGTGGTCAAAGTGGGTTCGGTTACCGTCCTGC
HO-1AAGCCGAGAATGCTGAGTTCAGCCGTGTAGATATGGTACAAGGA
PRDX1AATGCAAAAATTGGGTATCCTGCCGTGGGACACACAAAAGTAAAGT
TXN1CATGCCGACCTTCCAGTTTTATTTCCTTGTTAGCACCGGAGA
GCLCGGGGTGACGAGGTGGAGTAGTTGGGGTTTGTCCTCTCCC
GAPDHAGGTCGGTGTGAACGGATTTGTGTAGACCATGTAGTTGAGGTCA
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动物双歧杆菌CCFM1155灭活菌体调控Nrf2信号通路缓解小鼠皮肤衰老
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崔皓博 , 唐鑫 , 毛丙永 , 张秋香 , 翟齐啸 , 赵建新 , 陈卫 , 崔树茂
微生物学报 | 研究报告 2026,66(8): 3953-3964
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微生物学报 |研究报告 2026 , 66 (8) : 3953 -3964
动物双歧杆菌CCFM1155灭活菌体调控Nrf2信号通路缓解小鼠皮肤衰老
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崔皓博, 唐鑫 , 毛丙永, 张秋香, 翟齐啸, 赵建新, 陈卫, 崔树茂
作者信息
  • 江南大学 食品学院，江苏 无锡
通讯作者:
唐鑫
作者简介:

作者贡献声明

崔皓博：实验规划、数据整理与分析、文章撰写；唐鑫：实验指导、论文修改；毛丙永：文章构思；张秋香：文章讨论；翟齐啸：提供建议；赵建新：提供资源；陈卫：稿件审阅；崔树茂：监督管理。

Inactivated cells of Bifidobacterium animalis CCFM1155 modulate the Nrf2 signaling pathway to alleviate skin aging in mice
Haobo CUI, Xin TANG , Bingyong MAO, Qiuxiang ZHANG, Qixiao ZHAI, Jianxin ZHAO, Wei CHEN, 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.20260161
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【目的】 探究能够提高人永生化角质形成细胞(human immortalized epidermal cells, HaCaT)核转录因子E2相关因子2 (nuclear factor erythroid 2-related factor 2, Nrf2)表达量的动物双歧杆菌CCFM1155，口服其制备的后生元调控皮肤Nrf2信号通路并缓解皮肤衰老。 【方法】 通过每日皮下注射500 mg/kg D-半乳糖诱导皮肤衰老小鼠模型(n=6)，并分别灌胃5×109 CFU等量的动物双歧杆菌CCFM1155灭活菌体和浓缩上清发酵液，8周后检测小鼠皮肤衰老状况。 【结果】 动物双歧杆菌CCFM1155后生元中发挥主要抗皮肤衰老作用的成分为灭活菌体，而浓缩上清发酵液组未表现出明显改善效果。与模型组相比，口服CCFM1155灭活菌体组(1155B)能显著改善衰老小鼠皮肤组织结构，显著增加表皮厚度(P<0.05)与真皮胶原纤维密度(P<0.01)，降低衰老标志蛋白p21的表达及β-半乳糖苷酶活性(P<0.001)，有效缓解皮肤衰老表型。此外，1155B组还显著提高了皮肤中总超氧化物歧化酶(P<0.05)、过氧化氢酶活性(P<0.01)及还原型谷胱甘肽含量(P<0.01)，降低丙二醛水平(P<0.01)，增强整体抗氧化能力。进一步机制研究表明，灭活菌体可上调Nrf2的mRNA (P<0.05)和蛋白表达(P<0.001)，并促进其下游抗氧化基因血红素加氧酶-1 (heme oxygenase-1, HO-1)、过氧化物还原酶1 (peroxiredoxin 1, Prdx1)、硫氧还蛋白1 (thioredoxin 1, TXN1)和谷氨酸半胱氨酸连接酶催化亚基(glutamate-cysteine ligase catalytic subunit, GCLC)的转录，从而激活Nrf2下游信号通路。 【结论】 本研究初步证实动物双歧杆菌CCFM1155灭活菌体可通过激活Nrf2信号通路缓解皮肤衰老。该研究为后生元调控Nrf2通路提供了新视角，为开发安全有效的抗皮肤衰老产品奠定了理论基础。

动物双歧杆菌CCFM1155  /  Nrf2  /  皮肤抗衰老  /  后生元  /  D-半乳糖衰老

[Objective] To investigate the efficacy of postbiotics prepared with Bifidobacterium animalis CCFM1155, a strain previously identified for its ability to enhance nuclear factor erythroid 2-related factor 2 (Nrf2) secretion in human immortalized epidermal cells, in alleviating skin aging. [Methods] A mouse model of skin aging was established by daily subcutaneous injection of D-galactose (500 mg/kg) for eight weeks (n=6 per group). The model mice were then orally administered either heat-inactivated cells (1155B) or concentrated fermentation supernatant (1155T) of CCFM1155 cells at a dose equivalent to 5×109 CFU of live bacteria. [Results] The 1155B was the primary postbiotic component exerting anti-skin aging effects, whereas the 1155T group showed no significant effect. Compared with the model group, the 1155B treatment ameliorated skin aging phenotypes by improving the skin tissue structure, increasing the epidermal thickness (P<0.05) and dermal collagen fiber density (P<0.01), and decreasing the expression of the senescence marker protein p21 and the activity of senescence-associated β-galactosidase (P<0.001). In addition, the 1155B treatment enhanced the antioxidant defense system of the skin by elevating the activities of total superoxide dismutase (P<0.05) and catalase (P<0.01), as well as the content of reduced glutathione (P<0.01), while lowering the malondialdehyde level (P<0.01). Mechanism studies revealed that the 1155B upregulated both the mRNA (P<0.05) and protein (P<0.001) levels of Nrf2 in the skin. Consequently, it promoted the transcription of key downstream antioxidant molecules, including heme oxygenase-1, peroxiredoxin, thioredoxin 1, and the glutamate-cysteine ligase catalytic subunit, thereby activating the downstream signaling pathway of Nrf2. [Conclusion] The inactivated cells of B. animalis CCFM1155 alleviates skin aging by activating the Nrf2 signaling pathway. These findings offer a novel perspective on postbiotic regulation of the Nrf2 pathway and lay a theoretical foundation for developing safe and effective anti-skin aging products.

Bifidobacterium animalis CCFM1155  /  Nrf2  /  anti-skin aging  /  postbiotics  /  D-galactose-induced aging
崔皓博, 唐鑫, 毛丙永, 张秋香, 翟齐啸, 赵建新, 陈卫, 崔树茂. 动物双歧杆菌CCFM1155灭活菌体调控Nrf2信号通路缓解小鼠皮肤衰老. 微生物学报, 2026 , 66 (8) : 3953 -3964 . DOI: 10.13343/j.cnki.wsxb.20260161
Haobo CUI, Xin TANG, Bingyong MAO, Qiuxiang ZHANG, Qixiao ZHAI, Jianxin ZHAO, Wei CHEN, Shumao CUI. Inactivated cells of Bifidobacterium animalis CCFM1155 modulate the Nrf2 signaling pathway to alleviate skin aging in mice[J]. Acta Microbiologica Sinica, 2026 , 66 (8) : 3953 -3964 . DOI: 10.13343/j.cnki.wsxb.20260161
正文
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衰老是指生理功能随时间逐渐衰退的过程，其伴随着组织与器官的退行性变化，并最终导致机体死亡[1]。作为人体最大的器官，皮肤是抵御外界环境的第一道物理屏障。皮肤衰老表现为结构、细胞和分子层面的多重改变，以及衰老细胞的不断累积，这些细胞进一步诱发或加剧与衰老相关的功能障碍[2-3]。氧化应激是驱动皮肤衰老的核心因素之一[4]，其原因不仅包括活性氧(reactive oxygen species, ROS)生成增加，也与衰老过程中细胞自我修复能力下降密切相关[5]。
核转录因子E2相关因子2 (nuclear factor erythroid 2-related factor 2, Nrf2)是细胞抗氧化反应中的关键调控因子[6]。随着年龄增长，Nrf2表达水平下降，导致抗氧化能力减弱、氧化应激加剧，从而形成加速衰老的恶性循环[5]，因此提升Nrf2表达被视为干预衰老及相关疾病的重要策略。已有研究表明，内源性激活Nrf2通路或提高Nrf2蛋白表达可有效增强细胞抗氧化能力[4-5]。一些Nrf2激活剂，如萝卜硫素、姜黄素和二甲双胍等已被证实可增强Nrf2信号通路，被认为是延缓衰老及防治年龄相关疾病的潜在策略[7-8]。然而，尽管化学药物作用明确，却常伴随副作用，其在靶向干预的同时可能干扰正常生理功能，引发不良反应[9]。
后生元是一类新型微生态制剂，由灭活益生菌及其代谢产物组成，主要成分包括肽聚糖、胞外多糖、磷壁酸、细菌素和有机酸等[10]。与传统药物相比，后生元具有更高的安全性、稳定性，并更易于生产与贮存[11-12]。前期研究已筛选获得一株具有显著抗氧化潜力的动物双歧杆菌CCFM1155。体外实验结果表明，该菌后生元成分能显著提升H2O2诱导氧化损伤的人永生化角质形成细胞(human immortalized epidermal cells, HaCaT)存活率，并有效促进Nrf2的表达，从而缓解细胞氧化应激[13]。然而，其能否通过体内调控Nrf2的表达与激活，进而介导抗氧化信号通路以发挥皮肤抗衰老功效，仍有待进一步验证；其次，后生元发挥靶向调控Nrf2抗氧化通路作用的物质基础目前尚属未知。基于此，还需开展更深入的有效组分挖掘与系统性机制论证。本研究通过D-半乳糖诱导构建小鼠衰老模型，探讨口服动物双歧杆菌CCFM1155后生元对皮肤组织中Nrf2信号通路的调控作用，并评估其抗衰老潜力，以期为基于益生菌调控Nrf2通路及抗衰老功能的产品开发提供理论依据。
1 材料与方法
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1.1 材料
1.1.1 菌株
动物双歧杆菌CCFM1155，江南大学食品生物技术保藏中心。
1.1.2 主要试剂和仪器
D-半乳糖，Sigma-Aldrich公司；鼠源Nrf2试剂盒，上海酶联生物科技有限公司；RNA提取试剂盒、逆转录试剂盒、荧光定量试剂盒、SYBR Green Mix，南京诺唯赞生物科技股份有限公司；PCR相关引物，生工生物工程(上海)股份有限公司；过氧化氢酶(catalase, CAT)检测试剂盒、总超氧化物歧化酶(superoxide dismutase, SOD)测定试剂盒、还原型谷胱甘肽(glutathione, GSH)测定试剂盒、丙二醛(malondialdehyde, MDA)测定试剂盒，南京建成生物工程研究所有限公司；BCA蛋白浓度检测试剂盒，上海碧云天生物技术股份有限公司。
超净工作台，苏州安泰空气技术有限公司；隔水式恒温培养箱，上海森信实验仪器有限公司；Multiscan GO多功能酶标仪，赛默飞世尔科技(中国)有限公司；高通量组织研磨机，宁波新芝生物科技股份公司；组织切片电子扫描仪，3DHISTECH公司。实时荧光定量PCR仪、电泳仪，Bio-Rad公司。
1.1.3 培养基
简化MRS培养基(g/L)：酵母提取物5.0，蛋白胨5.0，葡萄糖15.0，磷酸氢二钠3.0，柠檬酸氢二铵3.0，磷酸氢二钾3.0，一水合硫酸锰0.05，七水合硫酸镁0.1，L-半胱氨酸1.0，吐温-80 1 mL。
1.2 后生元的制备
待动物双歧杆菌CCFM1155活化3代后，将其接种于简化MRS培养基中37 °C扩培24 h，测定活菌数后，8 000×g离心20 min。收集湿菌泥，加入生理盐水清洗2-3次，再加入适量无菌蒸馏水重悬，经巴氏杀菌灭活后计数活菌数，确保无活菌或少于103 CFU/g，随后进行冻干并称重；收集上清液，旋蒸浓缩20倍后于-20 °C保存。
后生元给药浓度的确定：测得动物双歧杆菌CCFM1155总活菌数为2×1012 CFU，总发酵液体积为1 000 mL。经冷冻干燥后获得菌粉总干重为1.43 g。由此计算，每毫克冻干菌粉相当于1.40×109 CFU当量(计算方法：2×1012 CFU/1.43×103 mg)；每毫升浓缩发酵液相当于4.0×1010 CFU当量(计算方法：2×1012 CFU/50 mL)。
1.3 动物实验
1.3.1 动物实验设计
本研究所有动物实验均获江南大学动物伦理委员会批准，编号为JN.No20240430b1200717[197]。8周龄BALB/c雄鼠，北京维通利华实验动物技术有限公司。36只雄性BALB/c小鼠(8周龄，SPF级，18-21 g)适应性饲养7 d后，随机分为6组(n=6)：空白对照(control)组、模型(model)组、培养基对照(med)组、阳性参照(pc)组、动物双歧杆菌CCFM1155灭活菌体(1155B)组和动物双歧杆菌CCFM1155发酵上清液(1155T)组。除control组外，其余各组小鼠每日背部皮下注射生理盐水配制的D-半乳糖(500 mg/kg)，连续造模8周。其间，1155B组小鼠每日灌胃5×109 CFU等量的灭活菌体；1155T组小鼠每日灌胃5×109 CFU等量的发酵上清液；med组每日灌胃等体积简化液体MRS培养基；pc组每2周连续5 d灌胃达沙替尼(5 mg/kg)和槲皮素(50 mg/kg)的10% PEG400溶液[14]；control组和model组小鼠每日灌胃等体积生理盐水。最后一次干预及造模结束后对小鼠背部进行剃毛，24 h后处死小鼠，立即取材。
1.3.2 小鼠皮肤组织病理学分析
收集小鼠背部皮肤组织，以4%多聚甲醛溶液固定24 h，梯度脱水后石蜡包埋。将石蜡块切成5 μm厚的切片，依据标准方法进行苏木精-伊红(hematoxylin-eosin, HE)染色和Masson染色。最后以中性树胶封片，待完全凝固后在数字组织切片扫描仪下拍照。所有组织学图像的定量分析均采用ImageJ软件完成。每份小鼠皮肤样本采集3个互不重叠的代表性视野，分别在10倍和20倍放大倍率下进行分析，取3个视野的平均值作为单个生物重复。每组随机选取3只小鼠(n=3)进行上述分析。
1.3.3 免疫组化分析
采用免疫组化法检测小鼠皮肤组织中p21和Nrf2的蛋白水平。将5 μm厚的背部皮肤石蜡切片置于60 °C烤箱中烤片120 min，二甲苯脱蜡3次(每次5 min)，梯度乙醇浸泡。切片浸入抗原修复液中，以3% H2O2溶液室温孵育10-20 min，PBS充分漂洗3次。3% BSA室温封闭2 h，滴加一抗4 °C过夜孵育，PBS充分洗涤3次(每次5 min)，彻底洗去未结合的一抗。滴加辣根过氧化物酶(horseradish peroxidase, HRP)标记的二抗，室温孵育1 h，PBS充分洗涤3次(每次5 min)。
1.3.4 酶联免疫吸附测定
取小鼠背部皮肤组织，加入预冷的放射免疫沉淀测定(radio immunoprecipitation assay, RIPA)缓冲液，于高通量组织研磨仪中破碎(65 Hz，5 s/次，循环10次)，4 °C、2 000 r/min离心10 min后取上清液，按照试剂盒说明书检测组织中Nrf2含量。测定结果以组织匀浆液总蛋白浓度进行归一化处理，总蛋白浓度采用BCA蛋白测定试剂盒测定。
1.3.5 抗氧化能力测定
取小鼠背部皮肤组织，加入预冷的PBS，于高通量组织研磨仪中破碎(65 Hz，5 s/次，循环10次)，4 °C、3 000 r/min离心10 min，取上清液，按照试剂盒说明书检测组织中SOD和CAT活力，以及MDA和GSH含量。抗氧化活性测定结果均以组织匀浆总蛋白浓度进行归一化处理，总蛋白浓度采用BCA蛋白测定试剂盒测定。
1.3.6 实时荧光定量PCR (qPCR)
将小鼠背部皮肤组织置于装有灭酶氧化锆珠的无酶离心管中，使用RNA提取试剂盒提取总RNA，并以逆转录试剂盒将1 μg总RNA逆转录为cDNA。以SYBR Green Mix制备PCR反应体系，在荧光定量基因扩增仪上检测各基因转录水平，采用2-ΔΔCt法计算目的基因相对表达量。PCR反应体系(10 µL)：Taq Pro Universal SYBR qPCR Master Mix 5 µL，正、反向引物(10 µmol/L)各0.25 µL，模板DNA (10 ng/µL) 1 µL，双蒸水3.5 µL。反应条件：95 ℃预变性5 min；95 ℃变性30 s，60 ℃退火30 s，72 ℃延伸2 min，共40个循环；72 ℃终止延伸5 min。引物序列如表1所示。
1.4 数据统计与分析
统计分析和绘图使用GraphPad Prism 9.0软件。实验结果重复3次，以平均值±标准误差表示。两组间比较采用双尾Student’s t检验；多组间比较采用单因素方差分析(one-way ANOVA)。
2 结果与分析
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2.1 口服动物双歧杆菌CCFM1155后生元对小鼠皮肤组织病理学的影响
各组小鼠背部皮肤组织HE染色结果如图1所示，表皮厚度测量结果如图2所示。control组小鼠皮肤表皮与真皮层交界清晰，表皮细胞排列紧密、形态正常，真皮乳头层和网状层清晰可辨，皮脂腺和毛囊分布均匀，成纤维细胞丰富且形态规则。相比之下，model组表现出明显的退行性病变，如表皮厚度显著减少(P<0.000 1)、真皮纤维组织排列疏松、表皮与真皮层连接受损，这与Liu等[15]的结果一致。与model组相比，动物双歧杆菌CCFM1155灭活菌体干预改善了上述退行性病变，表皮厚度显著增加(P<0.05)，真皮纤维组织排列趋于紧凑有序。Masson染色结果如图3所示，胶原蛋白相对定量结果如图4所示。control组真皮层中可见亮蓝色、密集的胶原蛋白束，排列整齐、厚度适中。与control组相比，model组胶原蛋白纤维松散且排列不均匀，部分区域出现纤维断裂或缺失。与model组相比，1155B组胶原蛋白纤维密度显著增加(P<0.01)，排列恢复规律。上述结果表明，口服动物双歧杆菌CCFM1155灭活菌体可有效改善皮肤衰老所致的退行性病变。 组和1155T组皮肤状态与model组相比无明显差异(P>0.05)，表明培养基及发酵上清液无显著改善作用。
2.2 口服动物双歧杆菌CCFM1155后生元对小鼠皮肤p21蛋白表达的影响
p21 (又名CDKN1A)是一种细胞周期蛋白依赖性激酶抑制剂，作为细胞衰老的核心调控蛋白和生物标志物[16]，可通过诱导皮肤细胞周期停滞及促进衰老相关分泌表型(senescence-associated secretory phenotype, SASP)的产生，降解胶原并引发炎症，直接驱动皮肤出现皱纹、松弛等衰老表征[3]。本研究通过免疫组化法检测了小鼠皮肤p21蛋白表达水平，结果如图5所示。control组皮肤中p21阳性染色较少，表皮层及毛囊等结构信号微弱，p21表达水平较低。与control组相比，D-半乳糖诱导后小鼠皮肤表皮层、毛囊及周围组织p21阳性染色显著增多，箭头所示区域可见明显棕色着色，表明p21表达显著上调。与model组相比，1155B组p21阳性染色明显减少，表皮及毛囊区域棕色信号减弱，接近control组水平，表明口服动物双歧杆菌CCFM1155灭活菌体可显著抑制皮肤p21蛋白表达，从而延缓皮肤衰老。Gerber等[17]，Samdavid等[18]研究表明，通过靶向p16、p21和p53等蛋白减少衰老细胞累积，可改善皮肤衰老状态并增强皮肤抗衰老能力。
2.3 口服动物双歧杆菌CCFM1155后生元对小鼠皮肤β-半乳糖苷酶活力的影响
β-半乳糖苷酶(senescence-associated β- galactosidase, SA-β-gal)活性升高是目前最广泛使用的细胞衰老生物标志物之一[19-20]，其在衰老皮肤细胞中活性显著增加，是衡量皮肤衰老程度的重要指标。如图6所示，与control组相比，model组SA-β-gal活力显著上调(P<0.000 1)，表明皮肤衰老程度较高。与model组相比，1155B组SA-β-gal活力下降了14.82% (P<0.001)，趋近control组水平。med组和1155T组SA-β-gal活力与model组相比无明显差异(P>0.05)。上述结果表明，口服动物双歧杆菌CCFM1155灭活菌体可显著降低小鼠皮肤SA-β-gal活力。综上所述，灭活菌体为动物双歧杆菌CCFM1155后生元的主要活性成分可增强皮肤健康状态，改善D-半乳糖诱导的皮肤退行性病变。
2.4 口服动物双歧杆菌CCFM1155后生元对小鼠皮肤抗氧化能力的影响
皮肤具有完善的抗氧化防御系统，包括SOD、CAT和谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)等酶类抗氧化剂，以及GSH、维生素C和维生素E等非酶类抗氧化剂[21]。如图7所示，与control组相比，D-半乳糖诱导的衰老皮肤SOD (P<0.000 1)和CAT (P<0.001)活性及GSH水平(P<0.001)均显著下降，表明皮肤遭受了氧化损伤。与model组相比，1155B组SOD活性提高了25.21% (P<0.05)，CAT活性提高了45.78% (P<0.01)，GSH水平提高了38.42% (P<0.01)。此外，MDA是脂质过氧化的主要产物，也是公认的氧化损伤生物标志物[22]。口服动物双歧杆菌CCFM1155灭活菌体可有效逆转D-半乳糖诱导的MDA升高(P<0.01)，与model组相比MDA水平降低了29.42% (P<0.01)。
2.5 口服动物双歧杆菌CCFM1155后生元对小鼠皮肤Nrf2/Keap1表达的影响
Kelch样ECH关联蛋白1 (Kelch like ECH associated protein 1, Keap1)是一种底物适配蛋白，能与细胞质中的Nrf2结合，在非应激条件下Nrf2经Keap1介导被蛋白酶体降解，表达水平较低；当受到亲电配体、ROS等刺激时Keap1发生构象改变，阻断Nrf2降解，Nrf2随之积累并发生核转位[23]。qPCR结果显示(图8)，与control组相比，Nrf2 mRNA相对表达量显著降低(P<0.001)；model组小鼠背部皮肤Keap1 mRNA相对表达量显著增加(P<0.001)，这与Li等[24]、Xia等[25]的研究结果一致。与model组相比，1155B组Nrf2 mRNA表达显著上调28.67% (P<0.05)，但Keap1 mRNA水平无明显差异(P>0.05)，表明其激活Nrf2通路的机制不依赖于Keap1下调。免疫组化结果如图9所示，相对定量结果如图10所示。control组表皮及真皮可见明显棕黄色阳性染色，Nrf2蛋白表达水平较高；model组棕黄色阳性染色显著减少(P<0.000 1)，表明D-半乳糖造模后Nrf2蛋白表达受到抑制；与model组相比，1155B组棕黄色阳性染色显著增多，Nrf2蛋白表达上调(P<0.001)，与qPCR结果一致。综上所述，口服动物双歧杆菌CCFM1155灭活菌体可提高皮肤中Nrf2的mRNA和蛋白表达水平，从而增强皮肤抗氧化能力。
2.6 口服动物双歧杆菌CCFM1155后生元对小鼠皮肤Nrf2下游信号通路的影响
当细胞遭受氧化应激时，Nrf2可转运至细胞核，结合抗氧化应答元件(antioxidant response element, ARE)后促进下游抗氧化酶的表达，从而保护细胞免受氧化损伤[26]。血红素加氧酶-1 (heme oxygenase-1, HO-1)的诱导被广泛认为是细胞对抗应激损伤的核心保护机制之一[27]；过氧化物还原酶1 (peroxiredoxin 1, Prdx1)可催化还原并清除细胞内的过氧化氢(H2O2)、过氧亚硝酸盐(ONOO⁻)和有机氢过氧化物，是维持细胞氧化还原平衡的关键酶[28]；硫氧还蛋白1 (thioredoxin 1, TXN1)是一种具有蛋白二硫键还原酶活性的氧化还原调节蛋白，通过催化二硫键还原维持细胞内氧化还原平衡[29]；谷氨酸-半胱氨酸连接酶催化亚基(glutamate-cysteine ligase catalytic subunit, GCLC)催化GSH生物合成的第一步反应，是该限速酶的催化核心亚基[30]。本研究通过qPCR检测了Nrf2通路下游靶基因的mRNA表达水平。如图11所示，D-半乳糖造模显著降低了各抗氧化酶mRNA的表达(P<0.000 1)。与model组相比，1155B组HO-1、Prdx1、TXN1和GCLC转录水平分别上调127.91% (P<0.001)、83.99% (P<0.000 1)、53.35% (P<0.001)和46.25% (P<0.01)。因此，口服动物双歧杆菌CCFM1155灭活菌体可调控Nrf2活化及核转位，并激活下游抗氧化信号通路。此外，Nrf2还可激活其他下游基因的转录与表达，这些基因共同参与抗氧化应激反应[30]。
3 讨论与结论
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本研究评估了动物双歧杆菌CCFM1155后生元对D-半乳糖诱导小鼠皮肤衰老的干预效果及其经Nrf2通路发挥抗氧化作用的功效。结果显示，其灭活菌体成分能够显著改善皮肤组织结构，增加表皮厚度与胶原蛋白密度，降低衰老相关蛋白p21的表达及SA-β-gal活性，有效缓解皮肤衰老表型。在抗氧化方面，口服动物双歧杆菌CCFM1155灭活菌体显著提高了皮肤组织中SOD和CAT活性及GSH含量，同时降低MDA水平，增强了皮肤的抗氧化防御能力。此外，口服动物双歧杆菌CCFM1155灭活菌体能够上调Nrf2的mRNA和蛋白表达，并促进其下游抗氧化基因HO-1、Prdx1、TXN1和GCLC的转录，从而激活Nrf2/ARE信号通路，增强机体对氧化应激的抵抗能力。与灭活菌体相比，发酵上清液未表现出明显的抗衰老作用及Nrf2调控效果。已有文献报道，不同益生菌来源的脂磷壁酸具有免疫调节、抗癌和抗氧化等活性[31]，乳杆菌来源的细胞蛋白具有抗氧化、抗炎和内皮保护特性[32]。基于此，本研究推测调控Nrf2抗氧化通路的关键功效物质可能来源于灭活菌体中的特定成分。综上所述，动物双歧杆菌CCFM1155灭活菌体而非发酵上清液，可通过调控Nrf2信号通路增强皮肤抗氧化能力，有效延缓皮肤衰老进程，为开发基于益生菌及其后生元的抗衰老产品提供了理论依据和实验支持。
本研究存在以下局限：首先，实验采用D-半乳糖诱导的小鼠衰老模型，小鼠与人类在皮肤结构、免疫系统和代谢特征上存在显著差异，研究结果向临床转化时需谨慎评估；其次，本研究缺乏萝卜硫素等已知Nrf2激活剂作为特异性阳性对照，尽管现有多维度数据初步构建了动物双歧杆菌CCFM1155灭活菌体激活Nrf2通路的证据链，但未来仍需引入特异性激活剂或抑制剂/敲除模型，并补充Nrf2核转位的直接证据，以进一步确证其靶向调控机制；此外，后生元中有效成分的鉴定尚不充分，灭活菌体中发挥作用的活性物质(如肽聚糖、胞外多糖或磷壁酸等)仍需进一步分离纯化与机制验证；最后，后生元的长期摄入稳定性、胃肠道存活率及生物利用度尚缺乏系统研究，上述因素可能影响其在人体中的实际功效发挥，未来需开展临床试验以验证其安全性与有效性。

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2026年第66卷第8期
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doi: 10.13343/j.cnki.wsxb.20260161
  • 接收时间：2026-02-27
  • 首发时间：2026-08-21
  • 出版时间：2026-08-04
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  • 收稿日期：2026-02-27
  • 录用日期：2026-04-16
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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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