The antiprotease Spink7 promotes inflammation resolution by modulating multiple proteases activities during wound healing

IF 7.9 1区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Clinical and Translational Medicine Pub Date : 2025-03-27 DOI:10.1002/ctm2.70291

Na Zhao, Guojian Wang, Shuang Long, Xiaofan Lv, Xinze Ran, Junping Wang, Yongping Su, Tao Wang

{"title":"The antiprotease Spink7 promotes inflammation resolution by modulating multiple proteases activities during wound healing","authors":"Na Zhao, Guojian Wang, Shuang Long, Xiaofan Lv, Xinze Ran, Junping Wang, Yongping Su, Tao Wang","doi":"10.1002/ctm2.70291","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Background</h3>\n \n <p>Effective control of inflammation is crucial for the healing of cutaneous wounds, but the molecular mechanisms governing inflammation resolution during wound closure are still not yet clear. Here, we describe a homeostatic mechanism that facilitates the inflammation resolution by timely regulating the targeted proteases activities through antiprotease Spink7 (serine peptidase inhibitor, kazal type 7).</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>The expression pattern of Spink7 was investigated by quantitative RT-PCR, immunohistochemistry (IHC) and in situ hybridization. In both Spink7 knockdown and knockout models, quantitative comparisons were made between the healing rate of wounds and histopathological morphometric analysis. Microarrays, multiple chemokine assays, IHC, immunofluorescence, protease activity measurement were performed to explore the underlying mechanisms of Spink7 knockout in impaired wound healing. Radiation-wound combined injury (R-W-CI) model was employed to evaluate the therapeutic effects of Spink7 manipulation.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>Our study demonstrates that Spink7 is significantly upregulated in the differentiated epidermal granular keratinocytes of proliferative phase during murine wound closure. Both local knockdown of Spink7 levels in wounds using siRNA gel and systemic knockout of Spink7 using KO mice resulted in delayed wound closure with sustained neutrophil infiltration. Loss of Spink7 leads to augmented inflammatory responses, increased production of multiple chemokines/cytokines, and impaired M2 polarization of macrophages in wound healing. Furthermore, loss of Spink7 results in elevated proteolytic activities of uPA, MMP2/9 and KLK5/7 in proliferative phase. However, inhibiting KLK5/7 downstream PAR2 activation exacerbates the phenotype of KO mice. In R-W-CI model, further significant induction of Spink7 is observed in wounds with insufficient inflammatory response. Local suppression of Spink7 promotes wound healing in the R-W-CI model by augmenting inflammation.</p>\n </section>\n \n <section>\n \n <h3> Conclusions</h3>\n \n <p>Maintaining an endogenous balance between Spink7 and its target proteases is a crucial checkpoint for regulating inflammation resolution during healing. Therefore, manipulating levels of Spink7 might be an effective treatment for impaired wounds caused by inflammatory dysregulation.</p>\n </section>\n </div>","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 4","pages":""},"PeriodicalIF":7.9000,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70291","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Clinical and Translational Medicine","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/ctm2.70291","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"MEDICINE, RESEARCH & EXPERIMENTAL","Score":null,"Total":0}

引用次数: 0

Abstract

Background

Effective control of inflammation is crucial for the healing of cutaneous wounds, but the molecular mechanisms governing inflammation resolution during wound closure are still not yet clear. Here, we describe a homeostatic mechanism that facilitates the inflammation resolution by timely regulating the targeted proteases activities through antiprotease Spink7 (serine peptidase inhibitor, kazal type 7).

Methods

The expression pattern of Spink7 was investigated by quantitative RT-PCR, immunohistochemistry (IHC) and in situ hybridization. In both Spink7 knockdown and knockout models, quantitative comparisons were made between the healing rate of wounds and histopathological morphometric analysis. Microarrays, multiple chemokine assays, IHC, immunofluorescence, protease activity measurement were performed to explore the underlying mechanisms of Spink7 knockout in impaired wound healing. Radiation-wound combined injury (R-W-CI) model was employed to evaluate the therapeutic effects of Spink7 manipulation.

Results

Our study demonstrates that Spink7 is significantly upregulated in the differentiated epidermal granular keratinocytes of proliferative phase during murine wound closure. Both local knockdown of Spink7 levels in wounds using siRNA gel and systemic knockout of Spink7 using KO mice resulted in delayed wound closure with sustained neutrophil infiltration. Loss of Spink7 leads to augmented inflammatory responses, increased production of multiple chemokines/cytokines, and impaired M2 polarization of macrophages in wound healing. Furthermore, loss of Spink7 results in elevated proteolytic activities of uPA, MMP2/9 and KLK5/7 in proliferative phase. However, inhibiting KLK5/7 downstream PAR2 activation exacerbates the phenotype of KO mice. In R-W-CI model, further significant induction of Spink7 is observed in wounds with insufficient inflammatory response. Local suppression of Spink7 promotes wound healing in the R-W-CI model by augmenting inflammation.

Conclusions

Maintaining an endogenous balance between Spink7 and its target proteases is a crucial checkpoint for regulating inflammation resolution during healing. Therefore, manipulating levels of Spink7 might be an effective treatment for impaired wounds caused by inflammatory dysregulation.

Abstract Image

查看原文本刊更多论文

抗蛋白酶Spink7在伤口愈合过程中通过调节多种蛋白酶的活性来促进炎症的消退

背景有效控制炎症对于皮肤伤口的愈合至关重要，但在伤口愈合过程中控制炎症消退的分子机制尚不清楚。本研究描述了一种体内平衡机制，通过抗蛋白酶Spink7（丝氨酸肽酶抑制剂，kazal 7型）及时调节靶向蛋白酶的活性，促进炎症的解决。方法采用定量RT-PCR、免疫组织化学（IHC）和原位杂交技术研究Spink7的表达模式。在Spink7敲低和敲除模型中，对创面愈合率和组织病理学形态学分析进行定量比较。通过微阵列、多种趋化因子测定、免疫组化、免疫荧光、蛋白酶活性测定来探索Spink7基因敲除在伤口愈合中的潜在机制。采用放射-创面复合损伤（R-W-CI）模型评价Spink7手法的治疗效果。结果我们的研究表明，Spink7在小鼠伤口愈合过程中增殖期分化的表皮颗粒角质形成细胞中表达显著上调。用siRNA凝胶局部敲除创面中的Spink7水平和用KO小鼠全身敲除Spink7都会导致伤口愈合延迟，并伴有中性粒细胞的持续浸润。Spink7的缺失导致炎症反应增强，多种趋化因子/细胞因子的产生增加，伤口愈合过程中巨噬细胞M2极化受损。此外，Spink7的缺失导致增殖期uPA、MMP2/9和KLK5/7蛋白水解活性升高。然而，抑制KLK5/7下游PAR2激活会加剧KO小鼠的表型。在R-W-CI模型中，在炎症反应不足的伤口中，Spink7进一步被显著诱导。在R-W-CI模型中，局部抑制Spink7通过增加炎症来促进伤口愈合。结论维持Spink7及其靶蛋白酶之间的内源性平衡是调节愈合过程中炎症消退的关键检查点。因此，控制Spink7的水平可能是一种有效的治疗炎症失调引起的伤口损伤的方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Clinical and Translational Medicine Multiple-

CiteScore

15.90

自引率

1.90%

发文量

450

审稿时长

4 weeks

期刊介绍： Clinical and Translational Medicine (CTM) is an international, peer-reviewed, open-access journal dedicated to accelerating the translation of preclinical research into clinical applications and fostering communication between basic and clinical scientists. It highlights the clinical potential and application of various fields including biotechnologies, biomaterials, bioengineering, biomarkers, molecular medicine, omics science, bioinformatics, immunology, molecular imaging, drug discovery, regulation, and health policy. With a focus on the bench-to-bedside approach, CTM prioritizes studies and clinical observations that generate hypotheses relevant to patients and diseases, guiding investigations in cellular and molecular medicine. The journal encourages submissions from clinicians, researchers, policymakers, and industry professionals.