Renliang Qu, Jingjing Liu, Lili Feng, Lianbing Li, Junwei Liu, Fengnan Sun, Lin Sun
{"title":"Down-regulation of <i>KLF9</i> ameliorates LPS-caused acute lung injury and inflammation in mice via reducing <i>GSDMD</i> expression.","authors":"Renliang Qu, Jingjing Liu, Lili Feng, Lianbing Li, Junwei Liu, Fengnan Sun, Lin Sun","doi":"10.1080/08916934.2022.2114465","DOIUrl":null,"url":null,"abstract":"<p><p>Acute lung injury (ALI) is considered as a severe respiratory disease with aggravated inflammatory responses. Krüppel-like factor 9 (<i>KLF9</i>), a member of KLF family, has been reported to be involved in inflammatory disorders. However, the effect of <i>KLF9</i> in ALI has not been elucidated. Here the present study was to clarify the role of <i>KLF9</i> and its mechanism in ALI. The ALI <i>in vitro</i> model was established with lipopolysaccharide (LPS)-treated RAW264.7 cells. Mice were injected with LPS to conduct an ALI <i>in vivo</i> model. The expression of <i>KLF9</i> and gasdermin D (<i>GSDMD</i>) was examined using quantitative reverse transcription-PCR, haematoxylin-eosin/immunohistochemistry staining and western blot assays. Enzyme-linked immunosorbent assay was employed to detect the levels of inflammatory cytokines. JASPAR database was used to predict the recognition motif of <i>KLF9</i>, and the relationship between <i>KLF9</i> and <i>GSDMD</i> was determined by luciferase reporter assay and chromatin immunoprecipitation analysis. The number of neutrophils in bronchoalveolar lavage fluid as well as the wet/dry weight ratio was caculated. The results showed that The expression of <i>KLF9</i> in lung was significantly increased in LPS-stimulated mice. Moreover, <i>KLF9</i> knockout relieved the lung injury in ALI mice. <i>GSDMD</i> is one of targets genes of the transcription factor <i>KLF9</i>. <i>KLF9</i> knockout induced a decreased expression of <i>GSDMD</i> in LPS-treated mice. Furthermore, in RAW264.7 cells after LPS administration, <i>KLF9</i> knockdown reduced the levels of inflammatory factors and suppressed the expression of <i>GSDMD</i>. In summarise, these findings exhibited that <i>KLF9</i> knockout could mitigate the lung injury and inflammatory responses in ALI mice by directly regulating <i>GSDMD</i>.</p>","PeriodicalId":3,"journal":{"name":"ACS Applied Electronic Materials","volume":null,"pages":null},"PeriodicalIF":4.3000,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"5","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Applied Electronic Materials","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1080/08916934.2022.2114465","RegionNum":3,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/8/21 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"ENGINEERING, ELECTRICAL & ELECTRONIC","Score":null,"Total":0}
引用次数: 5
Abstract
Acute lung injury (ALI) is considered as a severe respiratory disease with aggravated inflammatory responses. Krüppel-like factor 9 (KLF9), a member of KLF family, has been reported to be involved in inflammatory disorders. However, the effect of KLF9 in ALI has not been elucidated. Here the present study was to clarify the role of KLF9 and its mechanism in ALI. The ALI in vitro model was established with lipopolysaccharide (LPS)-treated RAW264.7 cells. Mice were injected with LPS to conduct an ALI in vivo model. The expression of KLF9 and gasdermin D (GSDMD) was examined using quantitative reverse transcription-PCR, haematoxylin-eosin/immunohistochemistry staining and western blot assays. Enzyme-linked immunosorbent assay was employed to detect the levels of inflammatory cytokines. JASPAR database was used to predict the recognition motif of KLF9, and the relationship between KLF9 and GSDMD was determined by luciferase reporter assay and chromatin immunoprecipitation analysis. The number of neutrophils in bronchoalveolar lavage fluid as well as the wet/dry weight ratio was caculated. The results showed that The expression of KLF9 in lung was significantly increased in LPS-stimulated mice. Moreover, KLF9 knockout relieved the lung injury in ALI mice. GSDMD is one of targets genes of the transcription factor KLF9. KLF9 knockout induced a decreased expression of GSDMD in LPS-treated mice. Furthermore, in RAW264.7 cells after LPS administration, KLF9 knockdown reduced the levels of inflammatory factors and suppressed the expression of GSDMD. In summarise, these findings exhibited that KLF9 knockout could mitigate the lung injury and inflammatory responses in ALI mice by directly regulating GSDMD.