Calcium-sensing receptor regulates the angiogenic differentiation of LPS-treated human dental pulp cells via the phosphoinositide 3-kinase/Akt pathway in vitro

IF 5.4 1区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE International endodontic journal Pub Date : 2024-07-30 DOI:10.1111/iej.14129

Ting Yang, Peiqi Liu, Zixin Qiu, Yuejiao Zhang, Shaofeng An

{"title":"Calcium-sensing receptor regulates the angiogenic differentiation of LPS-treated human dental pulp cells via the phosphoinositide 3-kinase/Akt pathway in vitro","authors":"Ting Yang, Peiqi Liu, Zixin Qiu, Yuejiao Zhang, Shaofeng An","doi":"10.1111/iej.14129","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Aim</h3>\n \n <p>The purpose of this study was to investigate the role of calcium-sensing receptor (CaSR) in the angiogenic differentiation of lipopolysaccharide (LPS)-treated human dental pulp cells (hDPCs).</p>\n </section>\n \n <section>\n \n <h3> Methodology</h3>\n \n <p>The LPS-induced hDPCs were cultured in the medium with different combinations of CaSR agonist R568 and antagonist Calhex231. The cell proliferation, migration, and angiogenic capacity were measured by Cell Counting Kit-8 (CCK-8), scratch wound healing, and tube formation assays, respectively. Enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), and western blot were conducted to determine the gene/protein expression of CaSR, inflammatory mediators, and angiogenic-associated markers. The activation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt) was assessed by western blot analysis.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>The cell proliferation was elevated in response to R568 or Calhex231 exposure, but an enhanced cell migration was only found in cultures supplemented with Calhex231. Furthermore, R568 was found to potentiate the formation of vessel-like structure, up-regulated the protein expression of tumour necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF), and stromal cell-derived factor (SDF)-1; comparable influences were also observed in R568-stimulated cells in the presence of PI3K inhibitor LY294002. In contrast, Calhex231 obviously inhibited the tube formation and VEGF protein level, whereas promoted the production of IL-6, TNF-α, and eNOS; however, in the presence of LY294002, Calhex231 showed a significant promotion on the protein expression of CaSR, VEGF, and SDF-1. In addition, R568 exhibited a promotive action on the Akt phosphorylation, which can be reversed by LY294002.</p>\n </section>\n \n <section>\n \n <h3> Conclusions</h3>\n \n <p>Our results demonstrated that CaSR can regulate the angiogenic differentiation of LPS-treated hDPCs with an involvement of the PI3K/Akt signalling pathway.</p>\n </section>\n </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":null,"pages":null},"PeriodicalIF":5.4000,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"International endodontic journal","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/iej.14129","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"DENTISTRY, ORAL SURGERY & MEDICINE","Score":null,"Total":0}

引用次数: 0

Abstract

Aim

The purpose of this study was to investigate the role of calcium-sensing receptor (CaSR) in the angiogenic differentiation of lipopolysaccharide (LPS)-treated human dental pulp cells (hDPCs).

Methodology

The LPS-induced hDPCs were cultured in the medium with different combinations of CaSR agonist R568 and antagonist Calhex231. The cell proliferation, migration, and angiogenic capacity were measured by Cell Counting Kit-8 (CCK-8), scratch wound healing, and tube formation assays, respectively. Enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), and western blot were conducted to determine the gene/protein expression of CaSR, inflammatory mediators, and angiogenic-associated markers. The activation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt) was assessed by western blot analysis.

Results

The cell proliferation was elevated in response to R568 or Calhex231 exposure, but an enhanced cell migration was only found in cultures supplemented with Calhex231. Furthermore, R568 was found to potentiate the formation of vessel-like structure, up-regulated the protein expression of tumour necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF), and stromal cell-derived factor (SDF)-1; comparable influences were also observed in R568-stimulated cells in the presence of PI3K inhibitor LY294002. In contrast, Calhex231 obviously inhibited the tube formation and VEGF protein level, whereas promoted the production of IL-6, TNF-α, and eNOS; however, in the presence of LY294002, Calhex231 showed a significant promotion on the protein expression of CaSR, VEGF, and SDF-1. In addition, R568 exhibited a promotive action on the Akt phosphorylation, which can be reversed by LY294002.

Conclusions

Our results demonstrated that CaSR can regulate the angiogenic differentiation of LPS-treated hDPCs with an involvement of the PI3K/Akt signalling pathway.

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

钙传感受体通过磷酸肌酸 3- 激酶/Akt途径调节体外经 LPS 处理的人牙髓细胞的血管生成分化。

目的：本研究旨在探究钙感受体（CaSR）在脂多糖（LPS）处理的人牙髓细胞（hDPCs）血管生成分化中的作用：方法：用不同组合的 CaSR 激动剂 R568 和拮抗剂 Calhex231 在培养基中培养 LPS 诱导的 hDPCs。细胞计数试剂盒-8（CCK-8）、划痕伤口愈合和血管形成试验分别测定了细胞的增殖、迁移和血管生成能力。酶联免疫吸附试验（ELISA）、实时定量聚合酶链反应（qRT-PCR）和免疫印迹法检测了 CaSR、炎症介质和血管生成相关标志物的基因/蛋白表达。免疫印迹分析评估了磷酸肌醇3-激酶（PI3K）和蛋白激酶B（Akt）的活化情况：结果：细胞增殖在暴露于 R568 或 Calhex231 的情况下得到提高，但只有在添加 Calhex231 的培养物中才发现细胞迁移增强。此外，还发现 R568 能促进血管样结构的形成，上调肿瘤坏死因子（TNF）-α、血管内皮生长因子（VEGF）和基质细胞衍生因子（SDF）-1 的蛋白表达；在 PI3K 抑制剂 LY294002 的存在下，R568 刺激的细胞也能观察到类似的影响。相比之下，Calhex231 明显抑制了管形成和血管内皮生长因子蛋白水平，同时促进了 IL-6、TNF-α 和 eNOS 的产生；然而，在 LY294002 的存在下，Calhex231 对 CaSR、血管内皮生长因子和 SDF-1 蛋白表达有显著促进作用。此外，R568 对 Akt 磷酸化有促进作用，而 LY294002 可逆转这种作用：我们的研究结果表明，CaSR 可调控经 LPS 处理的 hDPCs 的血管生成分化，PI3K/Akt 信号通路参与其中。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

International endodontic journal 医学-牙科与口腔外科

CiteScore

10.20

自引率

28.00%

发文量

195

审稿时长

4-8 weeks

期刊介绍： The International Endodontic Journal is published monthly and strives to publish original articles of the highest quality to disseminate scientific and clinical knowledge; all manuscripts are subjected to peer review. Original scientific articles are published in the areas of biomedical science, applied materials science, bioengineering, epidemiology and social science relevant to endodontic disease and its management, and to the restoration of root-treated teeth. In addition, review articles, reports of clinical cases, book reviews, summaries and abstracts of scientific meetings and news items are accepted. The International Endodontic Journal is essential reading for general dental practitioners, specialist endodontists, research, scientists and dental teachers.