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Effects of Porphyromonas gingivalis lipopolysaccharide on the expression of key genes involved in cholesterol metabolism in macrophages

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Cardiovascular diseases are positively correlated with periodontal disease. However, the molecular mechanisms linking atherosclerosis and periodontal infection are not clear. This study aimed to determine whether Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) altered the expression of genes regulating cholesterol metabolism in macrophages in the presence of low-density lipoprotein (LDL).

Material and methods: THP-1-derived macrophages were exposed to different concentrations (0.1, 1, 10 µg/ml) of LPS in the presence of 50 µg/ml native LDL. Macrophages were also incubated with 1 µg/ml LPS for varying times (0, 24, 48, or 72 h) in the presence of native LDL. Foam cell formation was determined by oil red O staining and cholesterol content quantification. CD36, lectin-like oxidized LDL receptor-1 (LOX-1), ATP-binding cassette G1 (ABCG1), and acetyl CoA acyltransferase 1 (ACAT1) expression levels were measured by western blot and qRT-PCR.

Results: Foam cell formation was induced in a time- and concentration-dependent manner as assessed by both morphological and biochemical criteria. Pg-LPS caused downregulation of CD36 and ABCG1 but upregulation of ACAT1, while LOX-1 expression was not affected (p = 0.137).

Conclusions: Pg-LPS appears to be an important link in the development of atherosclerosis by mechanisms targeting cholesterol homeostasis, namely, excess cholesterol ester formation via ACAT1 and reduced cellular cholesterol efflux via ABCG1.

No MeSH data available.


Measurement of time-dependent CD36 and LOX-1 expression in Pg-LPS-treated macrophages. Cells were treated with Pg-LPS (1.0 µg/ ml) and LDL (50 µg/ml) for increasing durations (0, 24, 48, or 72 h). A – qRT-PCR data of CD36 and LOX-1 mRNA expression are presented as fold changes relative to the untreated control (0 h). B – Western blot analysis of CD36 and LOX-1 protein expression. The protein levels were normalized to β-actin. *P < 0.05
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Figure 0004: Measurement of time-dependent CD36 and LOX-1 expression in Pg-LPS-treated macrophages. Cells were treated with Pg-LPS (1.0 µg/ ml) and LDL (50 µg/ml) for increasing durations (0, 24, 48, or 72 h). A – qRT-PCR data of CD36 and LOX-1 mRNA expression are presented as fold changes relative to the untreated control (0 h). B – Western blot analysis of CD36 and LOX-1 protein expression. The protein levels were normalized to β-actin. *P < 0.05

Mentions: Because of treatment with Pg-LPS (1 µg/ml) in the presence of LDL, we observed a significant time-dependent decrease in CD36 expression. CD36 mRNA expression dropped to 40% of that of the control (Figure 4 A), while the protein expression fell to less than 40% of that of the control (Figure 4 B). However, there was no significant time-dependent difference in LOX-1 expression, which was consistent with the results in cells treated with different concentrations of Pg-LPS.


Effects of Porphyromonas gingivalis lipopolysaccharide on the expression of key genes involved in cholesterol metabolism in macrophages
Measurement of time-dependent CD36 and LOX-1 expression in Pg-LPS-treated macrophages. Cells were treated with Pg-LPS (1.0 µg/ ml) and LDL (50 µg/ml) for increasing durations (0, 24, 48, or 72 h). A – qRT-PCR data of CD36 and LOX-1 mRNA expression are presented as fold changes relative to the untreated control (0 h). B – Western blot analysis of CD36 and LOX-1 protein expression. The protein levels were normalized to β-actin. *P < 0.05
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 0004: Measurement of time-dependent CD36 and LOX-1 expression in Pg-LPS-treated macrophages. Cells were treated with Pg-LPS (1.0 µg/ ml) and LDL (50 µg/ml) for increasing durations (0, 24, 48, or 72 h). A – qRT-PCR data of CD36 and LOX-1 mRNA expression are presented as fold changes relative to the untreated control (0 h). B – Western blot analysis of CD36 and LOX-1 protein expression. The protein levels were normalized to β-actin. *P < 0.05
Mentions: Because of treatment with Pg-LPS (1 µg/ml) in the presence of LDL, we observed a significant time-dependent decrease in CD36 expression. CD36 mRNA expression dropped to 40% of that of the control (Figure 4 A), while the protein expression fell to less than 40% of that of the control (Figure 4 B). However, there was no significant time-dependent difference in LOX-1 expression, which was consistent with the results in cells treated with different concentrations of Pg-LPS.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Cardiovascular diseases are positively correlated with periodontal disease. However, the molecular mechanisms linking atherosclerosis and periodontal infection are not clear. This study aimed to determine whether Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) altered the expression of genes regulating cholesterol metabolism in macrophages in the presence of low-density lipoprotein (LDL).

Material and methods: THP-1-derived macrophages were exposed to different concentrations (0.1, 1, 10 &micro;g/ml) of LPS in the presence of 50 &micro;g/ml native LDL. Macrophages were also incubated with 1 &micro;g/ml LPS for varying times (0, 24, 48, or 72 h) in the presence of native LDL. Foam cell formation was determined by oil red O staining and cholesterol content quantification. CD36, lectin-like oxidized LDL receptor-1 (LOX-1), ATP-binding cassette G1 (ABCG1), and acetyl CoA acyltransferase 1 (ACAT1) expression levels were measured by western blot and qRT-PCR.

Results: Foam cell formation was induced in a time- and concentration-dependent manner as assessed by both morphological and biochemical criteria. Pg-LPS caused downregulation of CD36 and ABCG1 but upregulation of ACAT1, while LOX-1 expression was not affected (p = 0.137).

Conclusions: Pg-LPS appears to be an important link in the development of atherosclerosis by mechanisms targeting cholesterol homeostasis, namely, excess cholesterol ester formation via ACAT1 and reduced cellular cholesterol efflux via ABCG1.

No MeSH data available.