牙龈卟啉单胞菌脂多糖对HUVECs中COX-2表达及信号通路的研究

doi:10.13701/j.cnki.kqyxyj.2022.05.010

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (5): 436-441.DOI: 10.13701/j.cnki.kqyxyj.2022.05.010

牙龈卟啉单胞菌脂多糖对HUVECs中COX-2表达及信号通路的研究

高源, 李佳, 李维善^*

佳木斯大学附属口腔医院牙周黏膜病科黑龙江佳木斯 154000

收稿日期:2021-12-27 出版日期:2022-05-28 发布日期:2022-05-20
通讯作者: *李维善,E-mail:weishanli666@126.com
作者简介:高源(1994~ ),女,河南南阳人,硕士在读,主要从事口腔牙周病与口腔黏膜病的研究工作。
基金资助:
省教育厅基本科研业务费基础研究项目(编号:2019-KYYWF-1360)

Study on COX-2 Expression and Signalling Pathway in HUVECs Induced by P.gingivalis Lipopolysaccharide

GAO Yuan, LI Jia, LI Weishan^*

Department of Periodontal Mucosal Disease, Stomatological Hospital, Jiamusi University, Jiamusi 154000, China

Received:2021-12-27 Online:2022-05-28 Published:2022-05-20

摘要/Abstract

摘要： 目的: 研究牙龈卟啉单胞菌脂多糖(P.gingivalis lipopolysaccharide,P.g-LPS)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)环氧合酶-2(cyclooxygenase-2,COX-2)表达的影响,以及p38MAPK和PI3K/Akt信号通路是否参与P.g-LPS对HUVECs中COX-2表达的调控。方法: 体外培养HUVECs,选择不同浓度的P.g-LPS(0.5、1.0、5.0、10.0 μg/mL)刺激6、18、24、36 h,采用细胞增殖检测试剂盒(CCK-8)检测P.g-LPS对HUVECs的细胞增殖影响。通过Western blot、实时荧光定量PCR(RT-qPCR)检测COX-2蛋白和基因的表达。体外培养HUVECs,不同浓度的P.g-LPS(0.5、1.0、5.0、10.0 μg/mL)刺激36 h后,添加p38MAPK和PI3K/Akt通路抑制剂SB203580和LY294002。采用Western blot法和RT-qPCR法检测各组COX-2蛋白和基因的变化情况。结果: P.g-LPS处理HUVECs后,与对照组比较,随着P.g-LPS浓度增加HUVECs增殖活力显著下降(P<0.05)。与对照组比较,P.g-LPS促进 HUVECs中COX-2的表达,且具有浓度依赖性(P<0.05)。同时,P.g-LPS激活了p38MAPK及PI3K/Akt信号通路,添加SB203580和LY294002抑制剂可以抑制P.g-LPS对COX-2的表达(P<0.05)。结论: P.g-LPS可明显抑制HUVECs的增殖,并且P.g-LPS可以通过激活p38MAPK及PI3K/Akt通路促进COX-2蛋白及基因的表达。

关键词: 牙龈卟啉单胞菌脂多糖, 人脐静脉内皮细胞, 环氧合酶-2, 牙周炎, 动脉粥样硬化

Abstract: Objective: To study the effect of P.gingivalis lipopolysaccharide on Cyclooxygenase-2 (Cox-2) expression in human umbilical vein endothelial cells (HUVECs), and investigate whether p38MAPK and PI3K/Akt signalling pathways involved in the regulation of COX-2 expression in HUVECs by P.g-LPS. Methods: HUVECs were cultured in vitro, and different concentrations of P.g-LPS (0.5, 1.0, 5.0, and 10.0 μg/mL) were used for stimulation for 6, 18, 24, and 36 hours. The effect of P.g-LPS on cell proliferation of HUVECs was detected by CCK-8. The expressions of COX-2 protein and gene in HUVECs were detected by Western blot and RT-qPCR. Additional, after stimulated with different concentrations of P.g-LPS(0.5, 1.0, 5.0, and 10.0 μg/mL)for 36 hours, HUVECs were added with pathway inhibitors SB203580 and LY294002. The expressions of COX-2 protein and gene in each group were detected by Western blot and RT-qPCR. Results: The proliferation activity of HUVECs decreased significantly with increasing P.g-LPS concentration (P<0.05). Compared with the control group, P.g-LPS promoted the expression of COX-2 in HUVECs in a concentration dependent manner (P<0.05). In addition, P.g-LPS activated the p38MAPK and PI3K/Akt signalling pathways. Addition of SB203580 and LY294002 inhibitors inhibited the expression of COX-2 by P.g-LPS (P<0.05). Conclusion: P.g-LPS can significantly inhibit the proliferation of HUVECs and may promote COX-2 protein and genes expression through activation of p38MAPK and PI3K/Akt pathways.

Key words: P.gingivalis lipopolysaccharide, human umbilical vein endothelial cells, cyclooxygenase-2, periodontitis, atherosclerosis

高源, 李佳, 李维善. 牙龈卟啉单胞菌脂多糖对HUVECs中COX-2表达及信号通路的研究[J]. 口腔医学研究, 2022, 38(5): 436-441.

GAO Yuan, LI Jia, LI Weishan. Study on COX-2 Expression and Signalling Pathway in HUVECs Induced by P.gingivalis Lipopolysaccharide[J]. Journal of Oral Science Research, 2022, 38(5): 436-441.

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牙龈卟啉单胞菌脂多糖对HUVECs中COX-2表达及信号通路的研究

Study on COX-2 Expression and Signalling Pathway in HUVECs Induced by P.gingivalis Lipopolysaccharide

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