Cargando…
Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways
BACKGROUND: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT’s mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Korean Society of Veterinary Science
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10556282/ https://www.ncbi.nlm.nih.gov/pubmed/38031648 http://dx.doi.org/10.4142/jvs.23148 |
_version_ | 1785116840964390912 |
---|---|
author | Shrestha, Saroj K Kim, Se-Woong Soh, Yunjo |
author_facet | Shrestha, Saroj K Kim, Se-Woong Soh, Yunjo |
author_sort | Shrestha, Saroj K |
collection | PubMed |
description | BACKGROUND: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT’s mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim was to investigate the impact of KT on vascular calcification for the treatment of cardiovascular disease. OBJECTIVES: Using diverse calcification media, we studied the effect of KT on VSMC calcification and the underlying mechanism of this effect. METHODS: VSMC was isolated from the 6 weeks ICR mice. Then VSMCs were treated with different concentrations of KT to check the cell viability. Alizarin red and von Kossa staining were carried out to examine the calcium deposition on VSMC. Thoracic aorta of 6 weeks mice were taken and treated with different concentrations of KT, and H and E staining was performed. Real-time polymerase chain reaction and western blot were performed to examine KT’s effect on VSMC mineralization. Calcium deposition on VSMC was examined with a calcium deposition quantification kit. RESULTS: Calcium deposition, Alizarin red, and von Kossa staining revealed that KT reduced inorganic phosphate-induced calcification phenotypes. KT also reduced Ca(++)-induced calcification by inhibiting genes that regulate osteoblast differentiation, such as runt-related transcription factor 2 (RUNX-2), SMAD family member 4, osterix, collagen 1α, and osteopontin. Also, KT repressed Ca(2+)-induced bone morphogenetic protein 2, RUNX-2, collagen 1α, osteoprotegerin, and smooth muscle actin protein expression. Likewise, Alizarin red and von Kossa staining showed that KT markedly decreased the calcification of ex vivo ring formation in the mouse thoracic aorta. CONCLUSIONS: This experiment demonstrated that KT decreases vascular calcification and may be developed as a new therapeutic treatment for vascular calcification and arteriosclerosis. |
format | Online Article Text |
id | pubmed-10556282 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | The Korean Society of Veterinary Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-105562822023-10-07 Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways Shrestha, Saroj K Kim, Se-Woong Soh, Yunjo J Vet Sci Original Article BACKGROUND: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT’s mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim was to investigate the impact of KT on vascular calcification for the treatment of cardiovascular disease. OBJECTIVES: Using diverse calcification media, we studied the effect of KT on VSMC calcification and the underlying mechanism of this effect. METHODS: VSMC was isolated from the 6 weeks ICR mice. Then VSMCs were treated with different concentrations of KT to check the cell viability. Alizarin red and von Kossa staining were carried out to examine the calcium deposition on VSMC. Thoracic aorta of 6 weeks mice were taken and treated with different concentrations of KT, and H and E staining was performed. Real-time polymerase chain reaction and western blot were performed to examine KT’s effect on VSMC mineralization. Calcium deposition on VSMC was examined with a calcium deposition quantification kit. RESULTS: Calcium deposition, Alizarin red, and von Kossa staining revealed that KT reduced inorganic phosphate-induced calcification phenotypes. KT also reduced Ca(++)-induced calcification by inhibiting genes that regulate osteoblast differentiation, such as runt-related transcription factor 2 (RUNX-2), SMAD family member 4, osterix, collagen 1α, and osteopontin. Also, KT repressed Ca(2+)-induced bone morphogenetic protein 2, RUNX-2, collagen 1α, osteoprotegerin, and smooth muscle actin protein expression. Likewise, Alizarin red and von Kossa staining showed that KT markedly decreased the calcification of ex vivo ring formation in the mouse thoracic aorta. CONCLUSIONS: This experiment demonstrated that KT decreases vascular calcification and may be developed as a new therapeutic treatment for vascular calcification and arteriosclerosis. The Korean Society of Veterinary Science 2023-08-28 /pmc/articles/PMC10556282/ /pubmed/38031648 http://dx.doi.org/10.4142/jvs.23148 Text en © 2023 The Korean Society of Veterinary Science https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0 (https://creativecommons.org/licenses/by-nc/4.0/) ) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Shrestha, Saroj K Kim, Se-Woong Soh, Yunjo Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways |
title | Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways |
title_full | Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways |
title_fullStr | Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways |
title_full_unstemmed | Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways |
title_short | Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways |
title_sort | kalkitoxin attenuates calcification of vascular smooth muscle cells via runx-2 signaling pathways |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10556282/ https://www.ncbi.nlm.nih.gov/pubmed/38031648 http://dx.doi.org/10.4142/jvs.23148 |
work_keys_str_mv | AT shresthasarojk kalkitoxinattenuatescalcificationofvascularsmoothmusclecellsviarunx2signalingpathways AT kimsewoong kalkitoxinattenuatescalcificationofvascularsmoothmusclecellsviarunx2signalingpathways AT sohyunjo kalkitoxinattenuatescalcificationofvascularsmoothmusclecellsviarunx2signalingpathways |