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Hyaluronic acid abrogates ethanol-dependent inhibition of collagen biosynthesis in cultured human fibroblasts.

Donejko M, Przylipiak A, Rysiak E, Miltyk W, Galicka E, Przylipiak J, Zaręba I, Surazynski A - Drug Des Devel Ther (2015)

Bottom Line: The application of HA has a protective effect on disturbances caused by the examined substances.It seems that regulatory mechanism of ethanol-induced collagen aberration take place at the level of collagen biosynthesis, since no effect of ethanol and HA was found on process of collagen degradation by MMP-2 and MMP-9.This study provides evidence that ethanol impairs collagen metabolism in human skin fibroblasts, leading to a significant decrease in the amount of produced protein.

View Article: PubMed Central - PubMed

Affiliation: Department of Esthetic Medicine, Medical University of Białystok, Białystok, Poland.

ABSTRACT

Introduction: The aim of the study was to evaluate the effect of ethanol on collagen biosynthesis in cultured human skin fibroblasts, and the role of hyaluronic acid (HA) in this process. Regarding the mechanism of ethanol action on human skin fibroblasts we investigated: expression of β1 integrin and insulin-like growth factor 1 receptor (IGF-IR), signaling pathway protein expression: mitogen-activated protein kinases (MAPKs), protein kinase B (Akt), nuclear factor kappa B (NF-κB) transcription factor, cytotoxicity assay and apoptosis, metalloproteinase activity, as well as the influence of HA on these processes.

Materials and methods: Collagen biosynthesis, activity of prolidase, DNA biosynthesis, and cytotoxicity were measured in confluent human skin fibroblast cultures that have been treated with 25, 50, and 100 mM ethanol and with ethanol and 500 µg/mL HA. Western blot analysis and zymography were performed to evaluate expression of collagen type I, β1 integrin receptor, IGF-IR, NF-κB protein, phospho-Akt protein, kinase MAPK, caspase 9 activity, and matrix metalloproteinases (MMP-9 and MMP-2).

Results: Ethanol in a dose-dependent manner lead to the impairment of collagen biosynthesis in fibroblast cultures through decreasing prolidase activity and expression of β1 integrin and IGF-IR. This was accompanied by an increased cytotoxicity, apoptosis and lowered expression of the signaling pathway proteins induced by β1 integrin and IGF-IR, that is, MAPK (ERK1/2) kinases. The lowered amount of synthesized collagen and prolidase activity disturbance may also be due to the activation of NF-κB transcription factor, which inhibits collagen gene expression. It suggests that the decrease in fibroblast collagen production may be caused by the disturbance in its biosynthesis but not degradation. The application of HA has a protective effect on disturbances caused by the examined substances. It seems that regulatory mechanism of ethanol-induced collagen aberration take place at the level of collagen biosynthesis, since no effect of ethanol and HA was found on process of collagen degradation by MMP-2 and MMP-9.

Conclusion: This study provides evidence that ethanol impairs collagen metabolism in human skin fibroblasts, leading to a significant decrease in the amount of produced protein. This mechanism probably is due to downregulation of prolidase activity, expression of β1 integrin and IGF-IR receptors, and the signaling pathway proteins induced by these receptors.

No MeSH data available.


Related in: MedlinePlus

Cytotoxicity in human skin fibroblasts (control) incubated for 24 hours with different concentrations of ethanol and HA.Notes: ANOVA test: *P<0.05, **P<0.01. Error bars represent ± standard deviation; n=3.Abbreviations: ANOVA, analysis of variance; HA, hyaluronic acid.
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f5-dddt-9-6225: Cytotoxicity in human skin fibroblasts (control) incubated for 24 hours with different concentrations of ethanol and HA.Notes: ANOVA test: *P<0.05, **P<0.01. Error bars represent ± standard deviation; n=3.Abbreviations: ANOVA, analysis of variance; HA, hyaluronic acid.

Mentions: The efficacy of HA in reducing the cytotoxicity of 25, 50, and 100 mM ethanol is shown in Figure 5. It has been shown that ethanol induced cytotoxicity in dose-dependent manner. The presence of ethanol inhibition of cell survival 20.16% (4.05%±SD; n=3), 35.18% (3.63%±SD; n=3), 56.03% (2.53%±SD; n=3) P<0.01, respectively, compared to the control value. The β1 integrin receptor plays a pivotal role in the control of collagen biosynthesis and prolidase activity. In this study, we found that ethanol decreased expression of β1 integrin receptor in dose-dependent manner. In cultured cells incubated with HA and ethanol, this effect was abrogated. The expression of integrin receptor in the cells treated with different concentration of ethanol and HA was found to be similar as in control cells (Figure 6A).


Hyaluronic acid abrogates ethanol-dependent inhibition of collagen biosynthesis in cultured human fibroblasts.

Donejko M, Przylipiak A, Rysiak E, Miltyk W, Galicka E, Przylipiak J, Zaręba I, Surazynski A - Drug Des Devel Ther (2015)

Cytotoxicity in human skin fibroblasts (control) incubated for 24 hours with different concentrations of ethanol and HA.Notes: ANOVA test: *P<0.05, **P<0.01. Error bars represent ± standard deviation; n=3.Abbreviations: ANOVA, analysis of variance; HA, hyaluronic acid.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664499&req=5

f5-dddt-9-6225: Cytotoxicity in human skin fibroblasts (control) incubated for 24 hours with different concentrations of ethanol and HA.Notes: ANOVA test: *P<0.05, **P<0.01. Error bars represent ± standard deviation; n=3.Abbreviations: ANOVA, analysis of variance; HA, hyaluronic acid.
Mentions: The efficacy of HA in reducing the cytotoxicity of 25, 50, and 100 mM ethanol is shown in Figure 5. It has been shown that ethanol induced cytotoxicity in dose-dependent manner. The presence of ethanol inhibition of cell survival 20.16% (4.05%±SD; n=3), 35.18% (3.63%±SD; n=3), 56.03% (2.53%±SD; n=3) P<0.01, respectively, compared to the control value. The β1 integrin receptor plays a pivotal role in the control of collagen biosynthesis and prolidase activity. In this study, we found that ethanol decreased expression of β1 integrin receptor in dose-dependent manner. In cultured cells incubated with HA and ethanol, this effect was abrogated. The expression of integrin receptor in the cells treated with different concentration of ethanol and HA was found to be similar as in control cells (Figure 6A).

Bottom Line: The application of HA has a protective effect on disturbances caused by the examined substances.It seems that regulatory mechanism of ethanol-induced collagen aberration take place at the level of collagen biosynthesis, since no effect of ethanol and HA was found on process of collagen degradation by MMP-2 and MMP-9.This study provides evidence that ethanol impairs collagen metabolism in human skin fibroblasts, leading to a significant decrease in the amount of produced protein.

View Article: PubMed Central - PubMed

Affiliation: Department of Esthetic Medicine, Medical University of Białystok, Białystok, Poland.

ABSTRACT

Introduction: The aim of the study was to evaluate the effect of ethanol on collagen biosynthesis in cultured human skin fibroblasts, and the role of hyaluronic acid (HA) in this process. Regarding the mechanism of ethanol action on human skin fibroblasts we investigated: expression of β1 integrin and insulin-like growth factor 1 receptor (IGF-IR), signaling pathway protein expression: mitogen-activated protein kinases (MAPKs), protein kinase B (Akt), nuclear factor kappa B (NF-κB) transcription factor, cytotoxicity assay and apoptosis, metalloproteinase activity, as well as the influence of HA on these processes.

Materials and methods: Collagen biosynthesis, activity of prolidase, DNA biosynthesis, and cytotoxicity were measured in confluent human skin fibroblast cultures that have been treated with 25, 50, and 100 mM ethanol and with ethanol and 500 µg/mL HA. Western blot analysis and zymography were performed to evaluate expression of collagen type I, β1 integrin receptor, IGF-IR, NF-κB protein, phospho-Akt protein, kinase MAPK, caspase 9 activity, and matrix metalloproteinases (MMP-9 and MMP-2).

Results: Ethanol in a dose-dependent manner lead to the impairment of collagen biosynthesis in fibroblast cultures through decreasing prolidase activity and expression of β1 integrin and IGF-IR. This was accompanied by an increased cytotoxicity, apoptosis and lowered expression of the signaling pathway proteins induced by β1 integrin and IGF-IR, that is, MAPK (ERK1/2) kinases. The lowered amount of synthesized collagen and prolidase activity disturbance may also be due to the activation of NF-κB transcription factor, which inhibits collagen gene expression. It suggests that the decrease in fibroblast collagen production may be caused by the disturbance in its biosynthesis but not degradation. The application of HA has a protective effect on disturbances caused by the examined substances. It seems that regulatory mechanism of ethanol-induced collagen aberration take place at the level of collagen biosynthesis, since no effect of ethanol and HA was found on process of collagen degradation by MMP-2 and MMP-9.

Conclusion: This study provides evidence that ethanol impairs collagen metabolism in human skin fibroblasts, leading to a significant decrease in the amount of produced protein. This mechanism probably is due to downregulation of prolidase activity, expression of β1 integrin and IGF-IR receptors, and the signaling pathway proteins induced by these receptors.

No MeSH data available.


Related in: MedlinePlus