A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling

Under normal condition, the endothelial cells form a compact monolayer to line the blood vessel. The integrity of this monolayer can be disrupted by lipopolysaccharide (LPS), where LPS could increase the permeability of the endothelium via dissociation of cell-to-cell junctions and rearrangement of...

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Main Authors: Chong, Yi Joong, Israf Ali, Daud Ahmad, Shaari, Khozirah, Tham, Chau Ling
Format: Article
Language:English
Published: Microscopy Society Malaysia 2015
Online Access:http://psasir.upm.edu.my/id/eprint/57679/
http://psasir.upm.edu.my/id/eprint/57679/1/A%20phloroglucinol%20from%20Melicope%20ptelefolia%20preserves%20barrier%20integrity%20of%20lipopolysaccharide-induced%20endothelial%20cells%20study%20on%20junctional%20proteins%20and%20cytoskeleton%20remodeling.pdf
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author Chong, Yi Joong
Israf Ali, Daud Ahmad
Shaari, Khozirah
Tham, Chau Ling
author_facet Chong, Yi Joong
Israf Ali, Daud Ahmad
Shaari, Khozirah
Tham, Chau Ling
author_sort Chong, Yi Joong
building UPM Institutional Repository
collection Online Access
description Under normal condition, the endothelial cells form a compact monolayer to line the blood vessel. The integrity of this monolayer can be disrupted by lipopolysaccharide (LPS), where LPS could increase the permeability of the endothelium via dissociation of cell-to-cell junctions and rearrangement of cytoskeleton. Prolonged LPS-induced inflammation may lead to excessive plasma leakage and leukocyte migration, leading to fatal outcomes such as shock and multiple organ dysfunctions. Control on this hallmark of inflammation was suggested as a mean of therapeutic interference to improve inflammation. In this study, we investigated the capability of tHGA on maintaining barrier integrity of LPS-induced endothelial cells. tHGA is a phloroglucinol found in Melicope ptelefolia. The compound was previously proven to have strong therapeutic potential with anti-asthma, anti cancer and anti-inflammatory properties. In murine experimental asthma model, tHGA has been shown to inhibit cytokine secretion and leukocyte infiltration, both of which are hallmarks of LPS-induced inflammation. Therefore, it is highly probable that tHGA may have anti-inflammatory effects on LPS-induced inflammation, Human Umbilical Vein Endothelial Cells (HUVECs) were used as an in vitro representative of endothelial cells. HUVECs were seeded to confluence in cell culture insert, where the cells were pretreated with tHGA and induced with LPS. Tracer dyes or monocytes were then allowed to pass the endothelium to examine severity of vascular leakage. To examine F-actin rearrangement, pretreated HUVECs were stained with Alexa-Fluur 488 Phalloidin prior to microscopic examination. The effect of tHGA on preserving junctional protein expression were examined via Western Blot. tHGA reduced permeability and monocyte transendothelial migration at 5µM and 20µM. Furthermore, tHGA inhibited LPS-induced cytoskeletal rearrangement of HUVECs. Additionally, tHGA is able to preserve the protein expression of F-cadherin and Occludin. tHGA is able to preserve barrier integrity of LPS-induced endothelial cells via preservation of actin cytoskeleton and junctional proteins.
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institution Universiti Putra Malaysia
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spelling upm-576792019-10-04T00:23:35Z http://psasir.upm.edu.my/id/eprint/57679/ A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling Chong, Yi Joong Israf Ali, Daud Ahmad Shaari, Khozirah Tham, Chau Ling Under normal condition, the endothelial cells form a compact monolayer to line the blood vessel. The integrity of this monolayer can be disrupted by lipopolysaccharide (LPS), where LPS could increase the permeability of the endothelium via dissociation of cell-to-cell junctions and rearrangement of cytoskeleton. Prolonged LPS-induced inflammation may lead to excessive plasma leakage and leukocyte migration, leading to fatal outcomes such as shock and multiple organ dysfunctions. Control on this hallmark of inflammation was suggested as a mean of therapeutic interference to improve inflammation. In this study, we investigated the capability of tHGA on maintaining barrier integrity of LPS-induced endothelial cells. tHGA is a phloroglucinol found in Melicope ptelefolia. The compound was previously proven to have strong therapeutic potential with anti-asthma, anti cancer and anti-inflammatory properties. In murine experimental asthma model, tHGA has been shown to inhibit cytokine secretion and leukocyte infiltration, both of which are hallmarks of LPS-induced inflammation. Therefore, it is highly probable that tHGA may have anti-inflammatory effects on LPS-induced inflammation, Human Umbilical Vein Endothelial Cells (HUVECs) were used as an in vitro representative of endothelial cells. HUVECs were seeded to confluence in cell culture insert, where the cells were pretreated with tHGA and induced with LPS. Tracer dyes or monocytes were then allowed to pass the endothelium to examine severity of vascular leakage. To examine F-actin rearrangement, pretreated HUVECs were stained with Alexa-Fluur 488 Phalloidin prior to microscopic examination. The effect of tHGA on preserving junctional protein expression were examined via Western Blot. tHGA reduced permeability and monocyte transendothelial migration at 5µM and 20µM. Furthermore, tHGA inhibited LPS-induced cytoskeletal rearrangement of HUVECs. Additionally, tHGA is able to preserve the protein expression of F-cadherin and Occludin. tHGA is able to preserve barrier integrity of LPS-induced endothelial cells via preservation of actin cytoskeleton and junctional proteins. Microscopy Society Malaysia 2015 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/57679/1/A%20phloroglucinol%20from%20Melicope%20ptelefolia%20preserves%20barrier%20integrity%20of%20lipopolysaccharide-induced%20endothelial%20cells%20study%20on%20junctional%20proteins%20and%20cytoskeleton%20remodeling.pdf Chong, Yi Joong and Israf Ali, Daud Ahmad and Shaari, Khozirah and Tham, Chau Ling (2015) A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling. Malaysian Journal of Microscopy, 11 (Supp 1). p. 21. ISSN 1823-7010
spellingShingle Chong, Yi Joong
Israf Ali, Daud Ahmad
Shaari, Khozirah
Tham, Chau Ling
A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling
title A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling
title_full A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling
title_fullStr A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling
title_full_unstemmed A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling
title_short A phloroglucinol from Melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling
title_sort phloroglucinol from melicope ptelefolia preserves barrier integrity of lipopolysaccharide-induced endothelial cells: study on junctional proteins and cytoskeleton remodeling
url http://psasir.upm.edu.my/id/eprint/57679/
http://psasir.upm.edu.my/id/eprint/57679/1/A%20phloroglucinol%20from%20Melicope%20ptelefolia%20preserves%20barrier%20integrity%20of%20lipopolysaccharide-induced%20endothelial%20cells%20study%20on%20junctional%20proteins%20and%20cytoskeleton%20remodeling.pdf