Fibulin-3调控组织金属蛋白酶抑制因子-3影响椎间盘髓核细胞衰老的机制研究
目的 探讨fibulin-3通过调控组织金属蛋白酶抑制因子-3(tissue inhibitor of metalloproteinase 3,TIMP-3)表达而影响椎间盘髓核细胞衰老的分子机制.方法 ①收集37例椎间盘术后患者的髓核组织及影像学资料,按照Pfirrmann分级评估椎间盘的退变程度,衰老相关β-半乳糖苷酶(senescence-associated β-galactosidase,SA-β-gal)染色法确定髓核细胞的衰老程度,Western blot和ELISA检测fibulin-3蛋白水平,探讨fibulin-3与椎间盘退变和髓核细胞衰老之间的关系;②体外培养人椎间盘髓核...
Saved in:
| Published in | 四川大学学报（医学版） Vol. 55; no. 5; pp. 1217 - 1225 |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
河南省洛阳正骨医院(河南省骨科医院)(洛阳 471002)%陆军军医大学新桥医院骨科(重庆 400037)%河南中医药大学(郑州 450046)
20.09.2024
|
| Subjects | |
| Online Access | Get full text |
| ISSN | 1672-173X |
| DOI | 10.12182/20240760604 |
Cover
| Abstract | 目的 探讨fibulin-3通过调控组织金属蛋白酶抑制因子-3(tissue inhibitor of metalloproteinase 3,TIMP-3)表达而影响椎间盘髓核细胞衰老的分子机制.方法 ①收集37例椎间盘术后患者的髓核组织及影像学资料,按照Pfirrmann分级评估椎间盘的退变程度,衰老相关β-半乳糖苷酶(senescence-associated β-galactosidase,SA-β-gal)染色法确定髓核细胞的衰老程度,Western blot和ELISA检测fibulin-3蛋白水平,探讨fibulin-3与椎间盘退变和髓核细胞衰老之间的关系;②体外培养人椎间盘髓核细胞(nucleus pulposus cells,NPCs),观察连续传代过程中NPCs的增殖(CCK8法)和衰老(SA-β-gal染色法)情况,检测fibulin-3的表达水平以及促炎症因子和基质蛋白酶的表达情况.通过添加外源性fibulin-3验证其对NPCs增殖和衰老的影响;③通过基因过表达技术验证fibulin-3对NPCs凋亡和增殖的作用,结合凋亡抑制剂进行双向验证;④利用生物信息学分析fibulin-3与TIMP家族的关系,通过过表达fibulin-3并沉默TIMP-3基因的实验,验证其在NPCs衰老中的作用.结果 ①在37例患者的椎间盘退变样本中,按Pfirrmann分级,退变等级越高,fibulin-3的表达越低.Spearman相关性分析显示椎间盘分级与NPCs的衰老等级负相关(r=-0.87,P＜0.001),与fibulin-3的表达量负相关(r=-0.79,P＜0.001);②随着NPCs传代次数增加,fibulin-3的表达逐渐降低,细胞增殖能力减弱,促炎症因子和基质蛋白酶的表达增高.添加外源性fibulin-3后,细胞形态和生长状态得到保持,细胞衰老显著抑制,促炎症因子和基质蛋白酶的表达明显降低;③基因过表达实验显示,fibulin-3减少了NPCs的凋亡,促进了细胞增殖,从而抑制了NPCs的衰老;④生物信息学分析显示fibulin-3与TIMP家族的TIMP-3存在显著关联.进一步实验验证发现,过表达fibulin-3能够增强TIMP-3的表达,而沉默TIMP-3基因后,fibulin-3对NPCs衰老的抑制作用显著减弱.这表明fibulin-3通过调控TIMP-3来抑制基质金属蛋白酶活性,影 |
|---|---|
| AbstractList | 目的 探讨fibulin-3通过调控组织金属蛋白酶抑制因子-3(tissue inhibitor of metalloproteinase 3,TIMP-3)表达而影响椎间盘髓核细胞衰老的分子机制.方法 ①收集37例椎间盘术后患者的髓核组织及影像学资料,按照Pfirrmann分级评估椎间盘的退变程度,衰老相关β-半乳糖苷酶(senescence-associated β-galactosidase,SA-β-gal)染色法确定髓核细胞的衰老程度,Western blot和ELISA检测fibulin-3蛋白水平,探讨fibulin-3与椎间盘退变和髓核细胞衰老之间的关系;②体外培养人椎间盘髓核细胞(nucleus pulposus cells,NPCs),观察连续传代过程中NPCs的增殖(CCK8法)和衰老(SA-β-gal染色法)情况,检测fibulin-3的表达水平以及促炎症因子和基质蛋白酶的表达情况.通过添加外源性fibulin-3验证其对NPCs增殖和衰老的影响;③通过基因过表达技术验证fibulin-3对NPCs凋亡和增殖的作用,结合凋亡抑制剂进行双向验证;④利用生物信息学分析fibulin-3与TIMP家族的关系,通过过表达fibulin-3并沉默TIMP-3基因的实验,验证其在NPCs衰老中的作用.结果 ①在37例患者的椎间盘退变样本中,按Pfirrmann分级,退变等级越高,fibulin-3的表达越低.Spearman相关性分析显示椎间盘分级与NPCs的衰老等级负相关(r=-0.87,P＜0.001),与fibulin-3的表达量负相关(r=-0.79,P＜0.001);②随着NPCs传代次数增加,fibulin-3的表达逐渐降低,细胞增殖能力减弱,促炎症因子和基质蛋白酶的表达增高.添加外源性fibulin-3后,细胞形态和生长状态得到保持,细胞衰老显著抑制,促炎症因子和基质蛋白酶的表达明显降低;③基因过表达实验显示,fibulin-3减少了NPCs的凋亡,促进了细胞增殖,从而抑制了NPCs的衰老;④生物信息学分析显示fibulin-3与TIMP家族的TIMP-3存在显著关联.进一步实验验证发现,过表达fibulin-3能够增强TIMP-3的表达,而沉默TIMP-3基因后,fibulin-3对NPCs衰老的抑制作用显著减弱.这表明fibulin-3通过调控TIMP-3来抑制基质金属蛋白酶活性,影 |
| Abstract_FL | Objective To investigate the effect of fibulin-3 on the senescence of intervertebral disc nucleus pulposus cells(NPCs)through the regulation of tissue inhibitor of metalloproteinases 3(TIMP-3)expression and to elucidate the molecular mechanisms involved.Methods 1).The nucleus pulposus tissues and imaging data of 37 patients who had undergone intervertebral disc surgery were collected.The degree of degeneration of the intervertebral discs were classified according to the Pfirrmann grading system.The senescence degree of NPCs was determined using senescence-associated β-galactosidase(SA-β-gal)staining.Fibulin-3 expression levels were determined using Western blot and ELISA.The relationship between fibulin-3 and disc degeneration and NPCs senescence was investigated.2).Human intervertebral disc NPCs were cultured in vitro.The proliferation and senescence of NPC across continuous passage were observed via CCK-8 assay and SA-β-gal staining,respectively.Fibulin-3 expression levels and the expression of inflammatory cytokines and matrix metalloproteinases were assessed.Exogenous fibulin-3 was added to verify its effect on the proliferation and senescence of NPCs.3).The effect of fibulin-3 on the apoptosis and proliferation of NPCs was verified through gene overexpression,which was used in combination with an apoptosis inhibitor for bidirectional verification.4).Bioinformatics analysis was performed to explore the relationship between fibulin-3 and the TIMP family.Experiments overexpressing fibulin-3 and silencing the TIMP-3 gene were performed to verify their role in NPCs senescence.Results 1).The intervertebral disc degeneration samples from 37 patients were classified according to the Pfirrmann grading system.The higher the degeneration grade,the lower fibulin-3 expression.Spearman correlation analysis showed that the disc grade was negatively correlated with the NPC senescence grade(r=-0.87,P＜0.001)and fibulin-3 expression(r=-0.79,P＜0.001).2).As the passage number of NPCs increased,fibulin-3 expression gradually decreased,cell proliferation ability weakened,and the expression of inflammatory cytokines and matrix metalloproteinases increased.After exogenous fibulin-3 was added,cell morphology and growth status were maintained,cell senescence was significantly inhibited,and the expression of inflammatory cytokines and matrix metalloproteinases was markedly reduced.3).Gene overexpression experiments showed that fibulin-3 reduced NPC apoptosis and promoted cell proliferation,thereby inhibiting NPC senescence.4).Bioinformatics analysis revealed a significant association between fibulin-3 and TIMP-3 of the TIMP family.Further experiments confirmed that overexpressing fibulin-3 enhanced TIMP-3 expression,while silencing the TIMP-3 gene significantly weakened the inhibitory effect of fibulin-3 on NPCs senescence.This indicates that,through regulating TIMP-3,fibulin-3 inhibits the activity of matrix metalloproteinases,affects the synthesis and degradation of the extracellular matrix,and ultimately inhibits NPCs senescence.Conclusion This study demonstrates that fibulin-3 plays a crucial role in inhibiting the senescence of intervertebral disc NPCs by regulating TIMP-3.The specific mechanisms involved are as follows,fibulin-3 upregulates TIMP-3 expression,inhibits matrix metalloproteinase activity,and reduces extracellular matrix degradation,thereby promoting extracellular matrix synthesis.Additionally,fibulin-3 inhibits NPCs senescence by reducing apoptosis and promoting cell proliferation.Therefore,fibulin-3 and TIMP-3 have potential therapeutic significance in maintaining intervertebral disc health and delaying degeneration. |
| Author | 王许可 王庆丰 孔军珂 张杨 罗刚 曹向阳 |
| AuthorAffiliation | 河南省洛阳正骨医院(河南省骨科医院)(洛阳 471002)%陆军军医大学新桥医院骨科(重庆 400037)%河南中医药大学(郑州 450046) |
| AuthorAffiliation_xml | – name: 河南省洛阳正骨医院(河南省骨科医院)(洛阳 471002)%陆军军医大学新桥医院骨科(重庆 400037)%河南中医药大学(郑州 450046) |
| Author_FL | LUO Gang WANG Xuke WANG Qingfeng CAO Xiangyang ZHANG Yang KONG Junke |
| Author_FL_xml | – sequence: 1 fullname: WANG Xuke – sequence: 2 fullname: ZHANG Yang – sequence: 3 fullname: LUO Gang – sequence: 4 fullname: KONG Junke – sequence: 5 fullname: CAO Xiangyang – sequence: 6 fullname: WANG Qingfeng |
| Author_xml | – sequence: 1 fullname: 王许可 – sequence: 2 fullname: 张杨 – sequence: 3 fullname: 罗刚 – sequence: 4 fullname: 孔军珂 – sequence: 5 fullname: 曹向阳 – sequence: 6 fullname: 王庆丰 |
| BookMark | eNotkE9LAkEchudgkJm3vkSHrd_M7O7MHkOyAqFLQTfZ2d0pS1ZokexWaBlFkhj0xxCDICIChZBI_Di7s_stkur0np7ngXcOpfyK7yG0gGEJE8zJMgGiAzPBBD2F0thkRMOM7syibBCUBADGQC3O0kjkS6JaLvkajQf1qPWqxg01bibNdjjsxd0r9TBJzkbRZTu8GIXdfvhxo9FwMgw719FLK7n7VN375L0T9b_U-Dyu9-LnQXxyqh4b0dP3lFD9W_U2mkcz0i4HXvZ_M2g7v7qVW9cKm2sbuZWCFmDQiUZdy6OGDlJ4FjhcMiYs4nCHcGEyaeuEGDYWtnSAOB6Xrs2YYVBOLGpIYXGgGbT45z2yfWn7u8X9SvXQnxaLe7XjA7dWE7-vGEAI_QEejncN |
| ContentType | Journal Article |
| Copyright | Copyright © Wanfang Data Co. Ltd. All Rights Reserved. |
| Copyright_xml | – notice: Copyright © Wanfang Data Co. Ltd. All Rights Reserved. |
| DBID | 2B. 4A8 92I 93N PSX TCJ |
| DOI | 10.12182/20240760604 |
| DatabaseName | Wanfang Data Journals - Hong Kong WANFANG Data Centre Wanfang Data Journals 万方数据期刊 - 香港版 China Online Journals (COJ) China Online Journals (COJ) |
| DatabaseTitleList | |
| DeliveryMethod | fulltext_linktorsrc |
| DocumentTitle_FL | Fibulin-3 Regulates Tissue Inhibitor of Metalloproteinases 3 to Inhibit Senescence in Intervertebral Disc Nucleus Pulposus Cells |
| EndPage | 1225 |
| ExternalDocumentID | hxykdxxb202405022 |
| GrantInformation_xml | – fundername: (河南省中医药科学研究专项课题); (洛阳市社会发展类公益专项) funderid: (河南省中医药科学研究专项课题); (洛阳市社会发展类公益专项) |
| GroupedDBID | -05 2B. 4A8 5XA 5XF 92I 93N ALMA_UNASSIGNED_HOLDINGS CCEZO CIEJG PSX RPM TCJ U1G U5O |
| ID | FETCH-LOGICAL-s1042-3d9e3540fbe90c8f77b92c8c28b67fa4225a1bafc02ce8fda7755382935fb9803 |
| ISSN | 1672-173X |
| IngestDate | Thu May 29 04:01:35 EDT 2025 |
| IsPeerReviewed | false |
| IsScholarly | true |
| Issue | 5 |
| Keywords | 髓核细胞 Intervertebral disc degeneration 椎间盘退变 组织金属蛋白酶抑制因子-3 Tissue inhibitor of metalloproteinases 3 Fibulin-3 Nucleus pulposus cells |
| Language | Chinese |
| LinkModel | OpenURL |
| MergedId | FETCHMERGED-LOGICAL-s1042-3d9e3540fbe90c8f77b92c8c28b67fa4225a1bafc02ce8fda7755382935fb9803 |
| PageCount | 9 |
| ParticipantIDs | wanfang_journals_hxykdxxb202405022 |
| PublicationCentury | 2000 |
| PublicationDate | 2024-09-20 |
| PublicationDateYYYYMMDD | 2024-09-20 |
| PublicationDate_xml | – month: 09 year: 2024 text: 2024-09-20 day: 20 |
| PublicationDecade | 2020 |
| PublicationTitle | 四川大学学报（医学版） |
| PublicationTitle_FL | Journal of Sichuan University(Medical Sciences) |
| PublicationYear | 2024 |
| Publisher | 河南省洛阳正骨医院(河南省骨科医院)(洛阳 471002)%陆军军医大学新桥医院骨科(重庆 400037)%河南中医药大学(郑州 450046) |
| Publisher_xml | – name: 河南省洛阳正骨医院(河南省骨科医院)(洛阳 471002)%陆军军医大学新桥医院骨科(重庆 400037)%河南中医药大学(郑州 450046) |
| SSID | ssib001103987 ssib000948209 ssib051370877 ssib000970525 ssib002039726 ssib001186719 ssib051810681 ssib009993695 ssib006703882 ssj0001742280 ssib002263209 |
| Score | 2.433511 |
| Snippet | 目的 探讨fibulin-3通过调控组织金属蛋白酶抑制因子-3(tissue inhibitor of metalloproteinase 3,TIMP-3)表达而影响椎间盘髓核细胞衰老的分子机制.方法... |
| SourceID | wanfang |
| SourceType | Aggregation Database |
| StartPage | 1217 |
| Title | Fibulin-3调控组织金属蛋白酶抑制因子-3影响椎间盘髓核细胞衰老的机制研究 |
| URI | https://d.wanfangdata.com.cn/periodical/hxykdxxb202405022 |
| Volume | 55 |
| hasFullText | 1 |
| inHoldings | 1 |
| isFullTextHit | |
| isPrint | |
| link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnR3batRANNT2xRdRVLxbxHkq0Vzn8jjZzVIEfWqhbyXZTawIK9gWap-UViuKxVLBS6VUEEREaEGKWPo5u9n9C8-ZZLNjt0gVljA7c-5nkswZcs4YxnWX0wTeg5FpJVFkegmPTRGnqZmmTiMSPPIShonCt-_Q8Unv1pQ_NTRyVftqaX4uvlFfPDSv5H-8Cn3gV8yS_QfPlkShA9rgX7iCh-F6JB_X7sX4IbnpkpCTwCIcGpTwkEhGQkaCgHCv14AegVdhk9AngU1EiFgChgKEEYIEVQUDo1TRkQUw56rHR2BpYUNWibCQrY9IgYISLuFVxJMeigCUBHBW_AFPcOyRAYIhjEUC3hONoiAgey6RtFEV7AGFbIUulR6UiAoJpCYRQzpCsZAgPtVX2oW88EMhgUhVCe4p2ygNJNUaSl3pk7BGggrSRy4VFK8PwwgXaiiHKbdQ1Eio7Ajih0ozwK4RWeuD-IiD1qMoiuQ6MthQsEIvIXUcNLSnRqhShSHZ_Byl3l6N4-GHJY5V3l3IInBIoNzDc9IMbYfWpOgSpCTQJUHuDJDHVe4BE-iaC5wV0oEg5K80S0SGxi3mzAEKokfkD-5jHsuL88KwX0BzqumrN0pvHPAhAFA1ZSjOHfThAPtBER1e3A9czQuYVpyOeapQUk-aQY09dK2sagw4DsnaIWIVDNzeDQdcwzHPx52i3u5bvhagzDFtpo6rLhcLeU3p4qHoa29-28lzkItVpO3k-fwDKxQ8MQGzsLC2IKNYuqq_Eiu_j51ZeHS_sbAQKygfltnHjBGHQXAwsB3pwQpej5cYnkvZj4fwQw8t_rJVOUstHoJhpu8v4OkNGj3KsFhUP_6BaNKl_UR-33YZFhTt_-e2RYtEdrXJzbDCoEpK7xmzSD5CM9zUjKByJptp1LyrLe8nThonirh8VOYP2VPG0OLMaSMuH7Cd7aX26pdsbznbW-murLV2NjsbL7P3-92nu-0Xa63nu62Nrdb316bb2t9prb9qf17tvv2Rbbzrfltvb_3M9p51ljY7n7Y7j59kH5bbH38BRrb1Jvu6e8aYrIUTlXGzOJTGnLUxldFtiAT3ytM4EVadp4zFwqnzusNjytII9PUjO47SuuXUE542IsZ8WFRCVOWnseCWe9YYbj5oJueMUQiOIz9NsGQqxIWpzxtpTC0WM0s4DP6eN64VNpkuXjqz0wMz48JRgC4ax_uPo0vG8NzD-eQyBFNz8RU1oX4DYwcPIw |
| linkProvider | National Library of Medicine |
| openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Fibulin-3%E8%B0%83%E6%8E%A7%E7%BB%84%E7%BB%87%E9%87%91%E5%B1%9E%E8%9B%8B%E7%99%BD%E9%85%B6%E6%8A%91%E5%88%B6%E5%9B%A0%E5%AD%90-3%E5%BD%B1%E5%93%8D%E6%A4%8E%E9%97%B4%E7%9B%98%E9%AB%93%E6%A0%B8%E7%BB%86%E8%83%9E%E8%A1%B0%E8%80%81%E7%9A%84%E6%9C%BA%E5%88%B6%E7%A0%94%E7%A9%B6&rft.jtitle=%E5%9B%9B%E5%B7%9D%E5%A4%A7%E5%AD%A6%E5%AD%A6%E6%8A%A5%EF%BC%88%E5%8C%BB%E5%AD%A6%E7%89%88%EF%BC%89&rft.au=%E7%8E%8B%E8%AE%B8%E5%8F%AF&rft.au=%E5%BC%A0%E6%9D%A8&rft.au=%E7%BD%97%E5%88%9A&rft.au=%E5%AD%94%E5%86%9B%E7%8F%82&rft.date=2024-09-20&rft.pub=%E6%B2%B3%E5%8D%97%E7%9C%81%E6%B4%9B%E9%98%B3%E6%AD%A3%E9%AA%A8%E5%8C%BB%E9%99%A2%28%E6%B2%B3%E5%8D%97%E7%9C%81%E9%AA%A8%E7%A7%91%E5%8C%BB%E9%99%A2%29%28%E6%B4%9B%E9%98%B3+471002%29%25%E9%99%86%E5%86%9B%E5%86%9B%E5%8C%BB%E5%A4%A7%E5%AD%A6%E6%96%B0%E6%A1%A5%E5%8C%BB%E9%99%A2%E9%AA%A8%E7%A7%91%28%E9%87%8D%E5%BA%86+400037%29%25%E6%B2%B3%E5%8D%97%E4%B8%AD%E5%8C%BB%E8%8D%AF%E5%A4%A7%E5%AD%A6%28%E9%83%91%E5%B7%9E+450046%29&rft.issn=1672-173X&rft.volume=55&rft.issue=5&rft.spage=1217&rft.epage=1225&rft_id=info:doi/10.12182%2F20240760604&rft.externalDocID=hxykdxxb202405022 |
| thumbnail_s | http://utb.summon.serialssolutions.com/2.0.0/image/custom?url=http%3A%2F%2Fwww.wanfangdata.com.cn%2Fimages%2FPeriodicalImages%2Fhxykdxxb%2Fhxykdxxb.jpg |