首页|TRIM21促进巨噬细胞M1极化加剧慢性根尖周炎进展的实验研究

TRIM21促进巨噬细胞M1极化加剧慢性根尖周炎进展的实验研究

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目的:探究三元基序蛋白21(TRIM21)在慢性根尖周炎(CAP)中的作用及机制.方法:收集人慢性根尖周炎和正常组织,RT-qPCR检测炎症因子(IL-1β、IL-6、TNF-α、TGF-β1)、破骨基因(TRAP、RANKL、CTSK)、巨噬细胞极化相关基因(CD86、iNOS、CD206、Arg1)及TRIM21的表达,抗酒石酸酸性磷酸酶(TRAP)染色检测破骨细胞,免疫组化染色检测TRIM21蛋白.以脂磷壁酸(LTA)或脂多糖(LPS)刺激Raw264.7细胞,并检测上述因子的表达,CCK-8法检测Raw264.7细胞增殖情况.提取野生型和TRIM21--小鼠骨髓来源巨噬细胞(BMDMs),免疫荧光染色鉴定.RT-qPCR验证LPS刺激的BMDMs中上述因子的表达,TRAP染色检测破骨细胞,免疫荧光染色检测巨噬细胞极化状态.结果:CAP组织中炎症因子、破骨基因、CD86、iNOS及TRIM21表达增加,CD206、Arg1表达减少,破骨细胞较正常组织多.LTA/LPS刺激促进Raw264.7细胞增殖,上述因子的表达情况与组织中的结果一致.LPS刺激后,TRIM21-/-的BMDM炎症程度较轻,破骨特异基因表达量较低,破骨细胞较少,M1极化程度较低.结论:TRIM21可能通过促进巨噬细胞M1极化从而促进CAP的进展.
An experimental study on TRIM21 promoting M1 polarization of macrophages to aggravate chronic apical periodontitis
Objective:To explore the role of tripartite motif-containing protein 21(TRIM21)in chronic apical periodontitis(CAP)and its potential mechanism.Methods:Human CAP tissue and normal periodontal tissue were collected.The expression of inflamma-tory factors(IL-1 β,IL-6,TNF-a,TGF-β1),osteoclast related genes(TRAP,RANKL,CTSK),macrophage polarization related genes(CD86,iNOS,CD206,Arg1)and TRIM21 were detected by RT-qPCR.TRIM21 protein was detected by immunohistochemical staining,and the osteoclasts was detected by tartrate resistant acid phosphatase(TRAP)staining.The inflammation cell model was es-tablished by stimulating Raw264.7 cells with lipoteichoic acid(LTA)or lipopolysaccharide(LPS),and the expression of the above factors was detected.The bone marrow-derived macrophages(BMDMs)extracted from wild-type and TRIM21-/-mice stimulated by LPS were used to verify the expression of the above factors by RT-qPCR,the osteoclasts were detected by TRAP staining,and the po-larization of macrophages was detected by immunofluorescence staining.Results:In CAP tissue the expression of inflammatory factors,osteoclast related genes,CD86,iNOS and TRIM21 increased,while CD206 and Arg1 decreased,and osteoclasts were more than that in normal tissue.The stimulation of LTA/LPS promoted the proliferation of Raw264.7 cells,and the expression of these factors in cells was consistent with that in tissues.After LPS stimulation,BMDMs of TRIM21-/-mice had lighter inflammation,lower expression of os-teoclast specific genes,fewer osteoclasts and lower M1 polarization than those of wild type mice.Conclusion:TRIM21 might promote the progress of CAP by promoting M1 polarization of macrophages.

TRIM21 proteinChronic apical periodontitisRaw264.7 cellsBone marrow-derived macrophagesPolarization

张静莹、刘小川、许晓芹、梁珮琪、王金斯、朱虹、王悦颖、吴柱国

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523710,广东医科大学附属东莞第一医院口腔科

TRIM21蛋白 慢性根尖周炎 Raw264.7细胞 骨髓来源巨噬细胞 极化

广东省基础与应用基础研究基金联合基金广东省普通高校重点领域项目广东医科大学学科建设项目广东医科大学学科建设项目广东医科大学大学生创新实验项目

2020B-15151200012020ZDZX-20134SG21015G4S-G21019GZY-DS004

2024

实用口腔医学杂志
第四军医大学口腔医学院

实用口腔医学杂志

CSTPCD北大核心
影响因子:0.942
ISSN:1001-3733
年,卷(期):2024.40(5)