硒代蛋氨酸通过NF-κB和MAPK通路抑制P.g-LPS诱导的RAW264.7细胞iNOS和COX-2表达

doi:10.13701/j.cnki.kqyxyj.2023.03.010

口腔医学研究 ›› 2023, Vol. 39 ›› Issue (3): 236-241.DOI: 10.13701/j.cnki.kqyxyj.2023.03.010

硒代蛋氨酸通过NF-κB和MAPK通路抑制P.g-LPS诱导的RAW264.7细胞iNOS和COX-2表达

张福¹, 周平¹, 向锋¹, 汪春艳¹, 李镭^2,3*

1.来凤县人民医院口腔科湖北恩施 445799;
2.湖北省口腔基础医学重点实验室-省部共建国家重点实验室培育基地,口腔生物医学教育部重点实验室(武汉大学) 湖北武汉 430079;
3.武汉大学口腔医院湖北武汉 430079

收稿日期:2022-08-22 出版日期:2023-03-28 发布日期:2023-03-21
通讯作者: * 李镭,E-mail:whusslilei@whu.edu.cn
作者简介:张福(1977~ ),男,湖北恩施人,本科,研究方向:牙周炎。
基金资助:
湖北省自然科学基金(编号:2020CFB455);恩施州科技计划(编号:JCY2019000018)

Selenomethionine Inhibited Expression of iNOS and COX-2 in P.g-LPS Induced RAW264.7 Cells through NF-κB and MAPK Pathways

ZHANG Fu¹, ZHOU Ping¹, XIANG Feng¹, WANG Chunyan¹, LI Lei^2,3*

1. Department of Stomatology, Laifeng County People's Hospital, Laifeng 445799, China;
2. The State Key Laboratory Breeding Base of Basic Science of Stomatology, Hubei Province & Key Laboratory of Oral Biomedicine (Wuhan University), Ministry of Education, Wuhan 430079, China;
3. Hospital of Stomatology, Wuhan University, Wuhan 430079, China

Received:2022-08-22 Online:2023-03-28 Published:2023-03-21

摘要/Abstract

摘要： 目的: 探讨硒代蛋氨酸(Selenomethionine,SeMet)对牙龈卟啉单胞菌脂多糖(Porphyromonas gingivalis lipopolysaccharide,P.g-LPS)诱导的RAW264.7细胞的作用和机制。方法: 使用P.g-LPS诱导小鼠巨噬细胞RAW264.7体外构建炎症细胞模型,CCK-8法检测不同浓度SeMet对RAW264.7细胞活性的影响。以不同浓度的SeMet(10、25、50 μmol/L)干预细胞1 h后,再使用P.g-LPS诱导细胞24 h。RT-PCR法检测诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和环氧合酶2(cyclooxygenase-2,COX-2)相关基因表达,ELISA法检测iNOS和COX-2相关蛋白分泌情况。Western blot检测NF-κB和MAPK信号通路相关蛋白表达情况。结果: 浓度低于50 μmol/L的SeMet对RAW264.7细胞活性没有显著影响。对RAW264.7使用SeMet预处理后,SeMet明显抑制P.g-LPS诱导的iNOS和COX-2相关基因表达和蛋白分泌增加,其作用与SeMet浓度有关,浓度越高,抑制作用越明显。其作用机制是通过抑制NF-κB通路的p65向核内转移,以及抑制MAPK通路的p38、ERK和JNK磷酸化发挥作用,并且其抑制作用呈浓度依赖性。结论: SeMet抑制P.g-LPS诱导的RAW264.7细胞iNOS和COX-2表达,其作用机制可能与SeMet抑制NF-κB p65核内转移和抑制MAPK通路p38、ERK和JNK的磷酸化有关。

关键词: 硒代蛋氨酸, 牙龈卟啉单胞菌脂多糖, 巨噬细胞, NF-κB信号通路

Abstract: Objective: To demonstrate the effects and mechanism of selenomethionine (SeMet) on porphyromonas gingivalis lipopolysaccharide (P.g-LPS) induced RAW264.7. Methods: P.g-LPS was used to stimulate the RAW264.7 cell line. CCK-8 was used to demonstrate the cell viability of RAW264.7 cells induced by SeMet on different concentration. After RAW264.7 was pretreated with SeMet (10,25,50μmol/L) for 1 h and then cultured with or without 1 μg/mL P.g-LPS for 24h, RT-PCR was used to clarify the gene expression of iNOS and COX-2, ELISA assay was used detect the protein secretion, and Western blot was used to determine the relative protein expression of NF-κB and MAPK signaling pathway. Results: SeMet lower than 50 μmol/L had no significant influence on RAW264.7 cell viability. After the RAW264.7 cells were pretreated with SeMet, the relative gene expression and protein secretion of iNOS and COX-2 were significantly inhibited by SeMet. The higher the concentration, the greater the inhibition. The mechanism was via inhibiting nuclear transcription of NF-κB p65 and phosphorylation of p38, ERK, and JNK of MAPK signaling pathway. The inhibiting effects were concentration-dependent. Conclusion: SeMet inhibited the expression of iNOS and COX-2 on RAW264.7 induced by P.g-LPS. The mechanism was related to inhibition of nuclear transcription of NF-κB p65 and phosphorylation of p38, ERK, and JNK of MAPK signaling pathway.

Key words: selenomethionine, Porphyromonas gingivalis lipopolysaccharide, macrophage, NF-κB signaling pathway

张福, 周平, 向锋, 汪春艳, 李镭. 硒代蛋氨酸通过NF-κB和MAPK通路抑制P.g-LPS诱导的RAW264.7细胞iNOS和COX-2表达[J]. 口腔医学研究, 2023, 39(3): 236-241.

ZHANG Fu, ZHOU Ping, XIANG Feng, WANG Chunyan, LI Lei. Selenomethionine Inhibited Expression of iNOS and COX-2 in P.g-LPS Induced RAW264.7 Cells through NF-κB and MAPK Pathways[J]. Journal of Oral Science Research, 2023, 39(3): 236-241.

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硒代蛋氨酸通过NF-κB和MAPK通路抑制P.g-LPS诱导的RAW264.7细胞iNOS和COX-2表达

Selenomethionine Inhibited Expression of iNOS and COX-2 in P.g-LPS Induced RAW264.7 Cells through NF-κB and MAPK Pathways

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