Microbiota control acute arterial inflammation and neointimal hyperplasia development after arterial injury

Kelly Wun, Betty R. Theriault, Joseph Pierre, Edmund B. Chen, Vanessa A. Leone, Katharine G. Harris, Liqun Xiong, Qun Jiang, Melanie Spedale, Owen M. Eskandari, Eugene B. Chang, Karen J. Ho

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background The microbiome has a functional role in a number of inflammatory processes and disease states. While neointimal hyperplasia development has been linked to inflammation, a direct role of the microbiota in neointimal hyperplasia has not yet been established. Germ-free (GF) mice are an invaluable model for studying causative links between commensal organisms and the host. We hypothesized that GF mice would exhibit altered neointimal hyperplasia following carotid ligation compared to conventionally raised (CONV-R) mice. Methods Twenty-week-old male C57BL/6 GF mice underwent left carotid ligation under sterile conditions. Maintenance of sterility was assessed by cultivation and 16S rRNA qPCR of stool. Neointimal hyperplasia was assessed by morphometric and histologic analysis of arterial sections after 28 days. Local arterial cell proliferation and inflammation was assessed by immunofluorescence for Ki67 and inflammatory cell markers at five days. Systemic inflammation was assessed by multiplex immunoassays of serum. CONV-R mice treated in the same manner served as the control cohort. GF and CONV-R mice were compared using standard statistical methods. Results All GF mice remained sterile during the entire study period. Twenty-eight days after carotid ligation, CONV-R mice had significantly more neointimal hyperplasia development compared to GF mice, as assessed by intima area, media area, intima+media area, and intima area/(intima+media) area. The collagen content of the neointimal lesions appeared qualitatively similar on Masson’s trichrome staining. There was significantly reduced Ki67 immunoreactivity in the media and adventitia of GF carotid arteries 5 days after ligation. GF mice also had increased arterial infiltration of anti-inflammatory M2 macrophages compared to CONV-R mouse arteries and a reduced proportion of mature neutrophils. GF mice had significantly reduced serum IFN-γ-inducible protein (IP)-10 and MIP-2 5 days after carotid ligation, suggesting a reduced systemic inflammatory response. Conclusions GF mice have attenuated neointimal hyperplasia development compared to CONV-R mice, which is likely related to altered kinetics of wound healing and acute inflammation. Recognizing the role of commensals in the regulation of arterial remodeling will provide a deeper understanding of the pathophysiology of restenosis and support strategies to treat or reduce restenosis risk by manipulating microbiota.

Original languageEnglish (US)
Article numbere0208426
JournalPloS one
Volume13
Issue number12
DOIs
StatePublished - Dec 1 2018

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Arteritis
Macrophages
Microbiota
Cell proliferation
Infiltration
hyperplasia
Hyperplasia
Statistical methods
Anti-Inflammatory Agents
Collagen
inflammation
Kinetics
mice
Wounds and Injuries
Proteins
Ligation
Inflammation
microbiome
Adventitia
aseptic conditions

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Wun, K., Theriault, B. R., Pierre, J., Chen, E. B., Leone, V. A., Harris, K. G., ... Ho, K. J. (2018). Microbiota control acute arterial inflammation and neointimal hyperplasia development after arterial injury. PloS one, 13(12), [e0208426]. https://doi.org/10.1371/journal.pone.0208426

Microbiota control acute arterial inflammation and neointimal hyperplasia development after arterial injury. / Wun, Kelly; Theriault, Betty R.; Pierre, Joseph; Chen, Edmund B.; Leone, Vanessa A.; Harris, Katharine G.; Xiong, Liqun; Jiang, Qun; Spedale, Melanie; Eskandari, Owen M.; Chang, Eugene B.; Ho, Karen J.

In: PloS one, Vol. 13, No. 12, e0208426, 01.12.2018.

Research output: Contribution to journalArticle

Wun, K, Theriault, BR, Pierre, J, Chen, EB, Leone, VA, Harris, KG, Xiong, L, Jiang, Q, Spedale, M, Eskandari, OM, Chang, EB & Ho, KJ 2018, 'Microbiota control acute arterial inflammation and neointimal hyperplasia development after arterial injury', PloS one, vol. 13, no. 12, e0208426. https://doi.org/10.1371/journal.pone.0208426
Wun, Kelly ; Theriault, Betty R. ; Pierre, Joseph ; Chen, Edmund B. ; Leone, Vanessa A. ; Harris, Katharine G. ; Xiong, Liqun ; Jiang, Qun ; Spedale, Melanie ; Eskandari, Owen M. ; Chang, Eugene B. ; Ho, Karen J. / Microbiota control acute arterial inflammation and neointimal hyperplasia development after arterial injury. In: PloS one. 2018 ; Vol. 13, No. 12.
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AU - Xiong, Liqun

AU - Jiang, Qun

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N2 - Background The microbiome has a functional role in a number of inflammatory processes and disease states. While neointimal hyperplasia development has been linked to inflammation, a direct role of the microbiota in neointimal hyperplasia has not yet been established. Germ-free (GF) mice are an invaluable model for studying causative links between commensal organisms and the host. We hypothesized that GF mice would exhibit altered neointimal hyperplasia following carotid ligation compared to conventionally raised (CONV-R) mice. Methods Twenty-week-old male C57BL/6 GF mice underwent left carotid ligation under sterile conditions. Maintenance of sterility was assessed by cultivation and 16S rRNA qPCR of stool. Neointimal hyperplasia was assessed by morphometric and histologic analysis of arterial sections after 28 days. Local arterial cell proliferation and inflammation was assessed by immunofluorescence for Ki67 and inflammatory cell markers at five days. Systemic inflammation was assessed by multiplex immunoassays of serum. CONV-R mice treated in the same manner served as the control cohort. GF and CONV-R mice were compared using standard statistical methods. Results All GF mice remained sterile during the entire study period. Twenty-eight days after carotid ligation, CONV-R mice had significantly more neointimal hyperplasia development compared to GF mice, as assessed by intima area, media area, intima+media area, and intima area/(intima+media) area. The collagen content of the neointimal lesions appeared qualitatively similar on Masson’s trichrome staining. There was significantly reduced Ki67 immunoreactivity in the media and adventitia of GF carotid arteries 5 days after ligation. GF mice also had increased arterial infiltration of anti-inflammatory M2 macrophages compared to CONV-R mouse arteries and a reduced proportion of mature neutrophils. GF mice had significantly reduced serum IFN-γ-inducible protein (IP)-10 and MIP-2 5 days after carotid ligation, suggesting a reduced systemic inflammatory response. Conclusions GF mice have attenuated neointimal hyperplasia development compared to CONV-R mice, which is likely related to altered kinetics of wound healing and acute inflammation. Recognizing the role of commensals in the regulation of arterial remodeling will provide a deeper understanding of the pathophysiology of restenosis and support strategies to treat or reduce restenosis risk by manipulating microbiota.

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