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Article: Lipopolysaccharide and hypoxia-induced HIF-1 activation in human gingival fibroblasts
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TitleLipopolysaccharide and hypoxia-induced HIF-1 activation in human gingival fibroblasts
 
AuthorsLi, JP2
Li, FYL1
Xu, A1
Cheng, B3
Tsao, SW1
Fung, ML1
Leung, WK2
 
KeywordsCell hypoxia
Chronic periodontitis
Hypoxia-inducible factor 1α subunit
Lipopolysaccharides
Toll-like receptor 4
Vascular endothelial growth factor A
 
Issue Date2012
 
PublisherAmerican Academy of Periodontology. The Journal's web site is located at http://www.perio.org
 
CitationJournal Of Periodontology, 2012, v. 83 n. 6, p. 816-824 [How to Cite?]
DOI: http://dx.doi.org/10.1902/jop.2011.110458
 
AbstractBackground: We previously reported that chronic periodontal inflammation causes the accumulation of the transcriptional activator hypoxia-inducible factor-1α (HIF-1 α) in human gingival fibroblasts (HGFs) in vivo. Here, evidence is provided that bacterial lipopolysaccharides (LPS) and cellular hypoxia, both associated with periodontitis, can individually, or in combination, lead to the accumulation and activation of HIF-1 in HGF in vitro. Methods: Primary gingival fibroblasts were cultured from human gingival biopsies. HIF-1 α peptide from HGFs treated with Escherichia coli LPS under normoxia or hypoxia was detected by nuclear protein extraction, immunoprecipitation, immunoblotting, and immunocytofluorescence. HIF-1 α transcripts were detected using reverse transcription polymerase chain reaction (PCR). The transcript expression levels of vascular endothelial growth factor-A (VEGF-A), a downstream gene of HIF-1α, were assessed by quantitative real-time PCR. Results: Two HIF-1 α splicing transcription variants were found to be constitutively expressed in HGFs. E. coli LPS induced a dose- and time-dependent nuclear accumulation of HIF-1 α peptide in HGFs. This accumulation could be attenuated by treatment with a Toll-like receptor 4 (TLR4)-neutraliz-ing antibody. Under hypoxia, LPS further increased HIF-1α accumulation in HGFs. VEGF-A transcript expression was upregulated by LPS under both normoxia and hypoxia but was downregulated by pretreatment with TLR4-neutralizing antibody or the specific HIF-1α inhibitor 3-(5'-hydroxymeth-yl-2'-furyl)-1-benzyl indazole. Conclusion: LPS induces the nuclear accumulation of HIF-1 α in HGFs and induces HIF-1 biologic activity under normoxia or hypoxia possibly through TLR4.
 
ISSN0022-3492
2013 Impact Factor: 2.565
2013 SCImago Journal Rankings: 1.143
 
DOIhttp://dx.doi.org/10.1902/jop.2011.110458
 
ISI Accession Number IDWOS:000305492800017
Funding AgencyGrant Number
University of Hong Kong200807176129
201007176307
200911159126
Funding Information:

This project was supported by the University of Hong Kong Small Project Funding Grants 200807176129 and 201007176307 and Seed Funding Grant 200911159126. The authors report no conflicts of interest related to this study.

 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorLi, JP
 
dc.contributor.authorLi, FYL
 
dc.contributor.authorXu, A
 
dc.contributor.authorCheng, B
 
dc.contributor.authorTsao, SW
 
dc.contributor.authorFung, ML
 
dc.contributor.authorLeung, WK
 
dc.date.accessioned2012-08-08T08:27:10Z
 
dc.date.available2012-08-08T08:27:10Z
 
dc.date.issued2012
 
dc.description.abstractBackground: We previously reported that chronic periodontal inflammation causes the accumulation of the transcriptional activator hypoxia-inducible factor-1α (HIF-1 α) in human gingival fibroblasts (HGFs) in vivo. Here, evidence is provided that bacterial lipopolysaccharides (LPS) and cellular hypoxia, both associated with periodontitis, can individually, or in combination, lead to the accumulation and activation of HIF-1 in HGF in vitro. Methods: Primary gingival fibroblasts were cultured from human gingival biopsies. HIF-1 α peptide from HGFs treated with Escherichia coli LPS under normoxia or hypoxia was detected by nuclear protein extraction, immunoprecipitation, immunoblotting, and immunocytofluorescence. HIF-1 α transcripts were detected using reverse transcription polymerase chain reaction (PCR). The transcript expression levels of vascular endothelial growth factor-A (VEGF-A), a downstream gene of HIF-1α, were assessed by quantitative real-time PCR. Results: Two HIF-1 α splicing transcription variants were found to be constitutively expressed in HGFs. E. coli LPS induced a dose- and time-dependent nuclear accumulation of HIF-1 α peptide in HGFs. This accumulation could be attenuated by treatment with a Toll-like receptor 4 (TLR4)-neutraliz-ing antibody. Under hypoxia, LPS further increased HIF-1α accumulation in HGFs. VEGF-A transcript expression was upregulated by LPS under both normoxia and hypoxia but was downregulated by pretreatment with TLR4-neutralizing antibody or the specific HIF-1α inhibitor 3-(5'-hydroxymeth-yl-2'-furyl)-1-benzyl indazole. Conclusion: LPS induces the nuclear accumulation of HIF-1 α in HGFs and induces HIF-1 biologic activity under normoxia or hypoxia possibly through TLR4.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationJournal Of Periodontology, 2012, v. 83 n. 6, p. 816-824 [How to Cite?]
DOI: http://dx.doi.org/10.1902/jop.2011.110458
 
dc.identifier.doihttp://dx.doi.org/10.1902/jop.2011.110458
 
dc.identifier.epage824
 
dc.identifier.hkuros198908
 
dc.identifier.isiWOS:000305492800017
Funding AgencyGrant Number
University of Hong Kong200807176129
201007176307
200911159126
Funding Information:

This project was supported by the University of Hong Kong Small Project Funding Grants 200807176129 and 201007176307 and Seed Funding Grant 200911159126. The authors report no conflicts of interest related to this study.

 
dc.identifier.issn0022-3492
2013 Impact Factor: 2.565
2013 SCImago Journal Rankings: 1.143
 
dc.identifier.issue6
 
dc.identifier.pmid22087807
 
dc.identifier.scopuseid_2-s2.0-84861792062
 
dc.identifier.spage816
 
dc.identifier.urihttp://hdl.handle.net/10722/154732
 
dc.identifier.volume83
 
dc.languageeng
 
dc.publisherAmerican Academy of Periodontology. The Journal's web site is located at http://www.perio.org
 
dc.publisher.placeUnited States
 
dc.relation.ispartofJournal of Periodontology
 
dc.relation.referencesReferences in Scopus
 
dc.subjectCell hypoxia
 
dc.subjectChronic periodontitis
 
dc.subjectHypoxia-inducible factor 1α subunit
 
dc.subjectLipopolysaccharides
 
dc.subjectToll-like receptor 4
 
dc.subjectVascular endothelial growth factor A
 
dc.titleLipopolysaccharide and hypoxia-induced HIF-1 activation in human gingival fibroblasts
 
dc.typeArticle
 
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<contributor.author>Cheng, B</contributor.author>
<contributor.author>Tsao, SW</contributor.author>
<contributor.author>Fung, ML</contributor.author>
<contributor.author>Leung, WK</contributor.author>
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<description.abstract>Background: We previously reported that chronic periodontal inflammation causes the accumulation of the transcriptional activator hypoxia-inducible factor-1&#945; (HIF-1 &#945;) in human gingival fibroblasts (HGFs) in vivo. Here, evidence is provided that bacterial lipopolysaccharides (LPS) and cellular hypoxia, both associated with periodontitis, can individually, or in combination, lead to the accumulation and activation of HIF-1 in HGF in vitro. Methods: Primary gingival fibroblasts were cultured from human gingival biopsies. HIF-1 &#945; peptide from HGFs treated with Escherichia coli LPS under normoxia or hypoxia was detected by nuclear protein extraction, immunoprecipitation, immunoblotting, and immunocytofluorescence. HIF-1 &#945; transcripts were detected using reverse transcription polymerase chain reaction (PCR). The transcript expression levels of vascular endothelial growth factor-A (VEGF-A), a downstream gene of HIF-1&#945;, were assessed by quantitative real-time PCR. Results: Two HIF-1 &#945; splicing transcription variants were found to be constitutively expressed in HGFs. E. coli LPS induced a dose- and time-dependent nuclear accumulation of HIF-1 &#945; peptide in HGFs. This accumulation could be attenuated by treatment with a Toll-like receptor 4 (TLR4)-neutraliz-ing antibody. Under hypoxia, LPS further increased HIF-1&#945; accumulation in HGFs. VEGF-A transcript expression was upregulated by LPS under both normoxia and hypoxia but was downregulated by pretreatment with TLR4-neutralizing antibody or the specific HIF-1&#945; inhibitor 3-(5&apos;-hydroxymeth-yl-2&apos;-furyl)-1-benzyl indazole. Conclusion: LPS induces the nuclear accumulation of HIF-1 &#945; in HGFs and induces HIF-1 biologic activity under normoxia or hypoxia possibly through TLR4.</description.abstract>
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Author Affiliations
  1. The University of Hong Kong Li Ka Shing Faculty of Medicine
  2. The University of Hong Kong
  3. Sun Yat-Sen University