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Functional Local Renin-Angiotensin System in Human and Rat Periodontal Tissue.

Santos CF, Morandini AC, Dionísio TJ, Faria FA, Lima MC, Figueiredo CM, Colombini-Ishikiriama BL, Sipert CR, Maciel RP, Akashi AP, Souza GP, Garlet GP, Rodini CO, Amaral SL, Becari C, Salgado MC, Oliveira EB, Matus I, Didier DN, Greene AS - PLoS ONE (2015)

Bottom Line: However, in inflamed tissue the immunoreactivity was greater for the AT1R compared to AT2R in fibroblasts.Ang 1-7 formation was significantly greater when human gingiva homogenates were incubated with chymostatin alone compared to incubation without any inhibitor, only captopril, or captopril and chymostatin.Furthermore, blocking AT1R and renin can significantly prevent periodontal bone loss induced by EP in rats.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.

ABSTRACT
The initiation or progression of periodontitis might involve a local renin-angiotensin system (RAS) in periodontal tissue. The aim of this study was to further characterize the local RAS in human and rat periodontal tissues between healthy and periodontally-affected tissue. Components of the RAS were investigated using in vitro, ex vivo and in vivo experiments involving both human and Wistar rat periodontium. Although not upregulated when challenged with P. gingivalis-lipopolysaccharide, human gingival and periodontal ligament fibroblasts expressed RAS components. Likewise, healthy and inflamed human gingiva expressed RAS components, some of which were shown to be functional, yet no differences in expression were found between healthy and diseased gingiva. However, in inflamed tissue the immunoreactivity was greater for the AT1R compared to AT2R in fibroblasts. When compared to healthy tissue, ACE activity was increased in human gingiva from volunteers with gingivitis. Human-gingiva homogenates generated Ang II, Ang 1-9 and Ang 1-7 when incubated with precursors. In gingiva homogenates, Ang II formation from Ang I was nearly abolished only when captopril and chymostatin were combined. Ang 1-7 formation was significantly greater when human gingiva homogenates were incubated with chymostatin alone compared to incubation without any inhibitor, only captopril, or captopril and chymostatin. In rat gingiva, RAS components were also found; their expression was not different between healthy and experimentally induced periodontitis (EP) groups. However, renin inhibition (aliskiren) and an AT1R antagonist (losartan) significantly blocked EP-alveolar-bone loss in rats. Collectively, these data are consistent with the hypothesis that a local RAS system is not only present but is also functional in both human and rat periodontal tissue. Furthermore, blocking AT1R and renin can significantly prevent periodontal bone loss induced by EP in rats.

No MeSH data available.


Related in: MedlinePlus

Renin-Angiotensin System Components in Human Fibroblasts.A to F) qPCR analysis of the following RAS components from human cultured gingival fibroblasts and periodontal ligament fibroblasts stimulated without or with LPS from P. gingivalis or E. coli: A) angiotensinogen (AGT), B) angiotensin converting enzyme (ACE), C) angiotensin converting enzyme 2 (ACE-2), D) angiotensin II receptor type 1 (AT1R), E) Mas receptor (MasR), and F) renin. Graphs displays relative expression levels of the target mRNA relative to RPL-13 mRNA from 3 donors in triplicate (n = 3). The means of gingival fibroblasts (open bars) and periodontal ligament fibroblasts (black bars) were compared using a 1-way ANOVA and Tukey’s test. N.D. = not detected. G) Fluorescent immunoreactivity of AT1R and MasR in gingival fibroblasts or periodontal ligament fibroblasts (portrayed in rows) when incubated with either cell culture medium as a negative control, interleukin-1β (IL-1β) as a positive control, lipopolysaccharide (LPS) from P. gingivalis, LPS from E. coli, or tissue without LPS as a negative control (portrayed in columns).
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pone.0134601.g004: Renin-Angiotensin System Components in Human Fibroblasts.A to F) qPCR analysis of the following RAS components from human cultured gingival fibroblasts and periodontal ligament fibroblasts stimulated without or with LPS from P. gingivalis or E. coli: A) angiotensinogen (AGT), B) angiotensin converting enzyme (ACE), C) angiotensin converting enzyme 2 (ACE-2), D) angiotensin II receptor type 1 (AT1R), E) Mas receptor (MasR), and F) renin. Graphs displays relative expression levels of the target mRNA relative to RPL-13 mRNA from 3 donors in triplicate (n = 3). The means of gingival fibroblasts (open bars) and periodontal ligament fibroblasts (black bars) were compared using a 1-way ANOVA and Tukey’s test. N.D. = not detected. G) Fluorescent immunoreactivity of AT1R and MasR in gingival fibroblasts or periodontal ligament fibroblasts (portrayed in rows) when incubated with either cell culture medium as a negative control, interleukin-1β (IL-1β) as a positive control, lipopolysaccharide (LPS) from P. gingivalis, LPS from E. coli, or tissue without LPS as a negative control (portrayed in columns).

Mentions: Beyond the demonstration of functional local RAS components in human gingiva, both cultured human gingival fibroblasts (GFs) and periodontal ligament fibroblasts (PLFs) expressed some components of the RAS under normal conditions and when stimulated with P. gingivalis and E. coli LPS. As indicated in Fig 4A–4F, the mRNA for AGT, ACE, ACE-2, AT1R, MasR and renin were detected in both GFs and PLFs, yet only the quantity of ACE mRNAs were significantly different between the fibroblast subtypes having greater expression in GFs when compared to PLFs (Fig 4B). Fibroblasts stimulated with LPS (10 μg/mL/24 h) did not alter the expression of RAS components compared to control (Fig 4A–4F). AT2R mRNA expression was found in fibroblasts stimulated with LPS but results were variable.


Functional Local Renin-Angiotensin System in Human and Rat Periodontal Tissue.

Santos CF, Morandini AC, Dionísio TJ, Faria FA, Lima MC, Figueiredo CM, Colombini-Ishikiriama BL, Sipert CR, Maciel RP, Akashi AP, Souza GP, Garlet GP, Rodini CO, Amaral SL, Becari C, Salgado MC, Oliveira EB, Matus I, Didier DN, Greene AS - PLoS ONE (2015)

Renin-Angiotensin System Components in Human Fibroblasts.A to F) qPCR analysis of the following RAS components from human cultured gingival fibroblasts and periodontal ligament fibroblasts stimulated without or with LPS from P. gingivalis or E. coli: A) angiotensinogen (AGT), B) angiotensin converting enzyme (ACE), C) angiotensin converting enzyme 2 (ACE-2), D) angiotensin II receptor type 1 (AT1R), E) Mas receptor (MasR), and F) renin. Graphs displays relative expression levels of the target mRNA relative to RPL-13 mRNA from 3 donors in triplicate (n = 3). The means of gingival fibroblasts (open bars) and periodontal ligament fibroblasts (black bars) were compared using a 1-way ANOVA and Tukey’s test. N.D. = not detected. G) Fluorescent immunoreactivity of AT1R and MasR in gingival fibroblasts or periodontal ligament fibroblasts (portrayed in rows) when incubated with either cell culture medium as a negative control, interleukin-1β (IL-1β) as a positive control, lipopolysaccharide (LPS) from P. gingivalis, LPS from E. coli, or tissue without LPS as a negative control (portrayed in columns).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526652&req=5

pone.0134601.g004: Renin-Angiotensin System Components in Human Fibroblasts.A to F) qPCR analysis of the following RAS components from human cultured gingival fibroblasts and periodontal ligament fibroblasts stimulated without or with LPS from P. gingivalis or E. coli: A) angiotensinogen (AGT), B) angiotensin converting enzyme (ACE), C) angiotensin converting enzyme 2 (ACE-2), D) angiotensin II receptor type 1 (AT1R), E) Mas receptor (MasR), and F) renin. Graphs displays relative expression levels of the target mRNA relative to RPL-13 mRNA from 3 donors in triplicate (n = 3). The means of gingival fibroblasts (open bars) and periodontal ligament fibroblasts (black bars) were compared using a 1-way ANOVA and Tukey’s test. N.D. = not detected. G) Fluorescent immunoreactivity of AT1R and MasR in gingival fibroblasts or periodontal ligament fibroblasts (portrayed in rows) when incubated with either cell culture medium as a negative control, interleukin-1β (IL-1β) as a positive control, lipopolysaccharide (LPS) from P. gingivalis, LPS from E. coli, or tissue without LPS as a negative control (portrayed in columns).
Mentions: Beyond the demonstration of functional local RAS components in human gingiva, both cultured human gingival fibroblasts (GFs) and periodontal ligament fibroblasts (PLFs) expressed some components of the RAS under normal conditions and when stimulated with P. gingivalis and E. coli LPS. As indicated in Fig 4A–4F, the mRNA for AGT, ACE, ACE-2, AT1R, MasR and renin were detected in both GFs and PLFs, yet only the quantity of ACE mRNAs were significantly different between the fibroblast subtypes having greater expression in GFs when compared to PLFs (Fig 4B). Fibroblasts stimulated with LPS (10 μg/mL/24 h) did not alter the expression of RAS components compared to control (Fig 4A–4F). AT2R mRNA expression was found in fibroblasts stimulated with LPS but results were variable.

Bottom Line: However, in inflamed tissue the immunoreactivity was greater for the AT1R compared to AT2R in fibroblasts.Ang 1-7 formation was significantly greater when human gingiva homogenates were incubated with chymostatin alone compared to incubation without any inhibitor, only captopril, or captopril and chymostatin.Furthermore, blocking AT1R and renin can significantly prevent periodontal bone loss induced by EP in rats.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.

ABSTRACT
The initiation or progression of periodontitis might involve a local renin-angiotensin system (RAS) in periodontal tissue. The aim of this study was to further characterize the local RAS in human and rat periodontal tissues between healthy and periodontally-affected tissue. Components of the RAS were investigated using in vitro, ex vivo and in vivo experiments involving both human and Wistar rat periodontium. Although not upregulated when challenged with P. gingivalis-lipopolysaccharide, human gingival and periodontal ligament fibroblasts expressed RAS components. Likewise, healthy and inflamed human gingiva expressed RAS components, some of which were shown to be functional, yet no differences in expression were found between healthy and diseased gingiva. However, in inflamed tissue the immunoreactivity was greater for the AT1R compared to AT2R in fibroblasts. When compared to healthy tissue, ACE activity was increased in human gingiva from volunteers with gingivitis. Human-gingiva homogenates generated Ang II, Ang 1-9 and Ang 1-7 when incubated with precursors. In gingiva homogenates, Ang II formation from Ang I was nearly abolished only when captopril and chymostatin were combined. Ang 1-7 formation was significantly greater when human gingiva homogenates were incubated with chymostatin alone compared to incubation without any inhibitor, only captopril, or captopril and chymostatin. In rat gingiva, RAS components were also found; their expression was not different between healthy and experimentally induced periodontitis (EP) groups. However, renin inhibition (aliskiren) and an AT1R antagonist (losartan) significantly blocked EP-alveolar-bone loss in rats. Collectively, these data are consistent with the hypothesis that a local RAS system is not only present but is also functional in both human and rat periodontal tissue. Furthermore, blocking AT1R and renin can significantly prevent periodontal bone loss induced by EP in rats.

No MeSH data available.


Related in: MedlinePlus