EBV LMP1对牙龈上皮细胞促炎因子的调控及机制研究

doi:10.13701/j.cnki.kqyxyj.2020.07.011

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (7): 654-657.DOI: 10.13701/j.cnki.kqyxyj.2020.07.011

EBV LMP1对牙龈上皮细胞促炎因子的调控及机制研究

刘芳芳¹, 邱忠营², 王艳丽^1*

1.西安市中心医院口腔科陕西西安 710003;
2.西安医学院基础医学部,陕西省脑疾病重点实验室陕西西安 710021

收稿日期:2019-11-21 出版日期:2020-07-28 发布日期:2020-07-24
通讯作者: 王艳丽,E-mail:xiatian1135@126.com
作者简介:刘芳芳(1987~),女,陕西西安人,硕士,主治医师,研究方向:口腔种植及牙周病学。
基金资助:
陕西省自然科学基础研究计划(编号:2018JQ8003)

Regulation and Mechanism of EBV LMP1 on Inflammatory Factors of Gingival Epithelial Cells

LIU Fangfang¹, QIU Zhongying², WANG Yanli^1*

1. Department of Stomatology, Xi'an Central Hospital, Xi'an 710003, China;
2. School of Basic Medical Sciences & Shanxi Key Laboratory of Brain Disorders, Xi'an Medical University, Xi'an 710021, China

Received:2019-11-21 Online:2020-07-28 Published:2020-07-24

摘要/Abstract

摘要： 目的: 探究EB病毒潜在膜蛋白1(Epstein-barr virus latent membrane protein 1, EBV LMP1)对牙龈上皮细胞促炎因子的调控及机制。方法: 0.05 μg和0.2 μg pSG-GFP-LMP1质粒转染Ca9-22细胞,分别于24 h和48 h后荧光显微镜下观察GFP在Ca9-22细胞中的表达情况,Real-time PCR和Western blot检测LMP1 mRNA和蛋白表达,确定转染是否成功。应用0.2 μg pSG-GFP-LMP1质粒转染Ca9-22细胞,分别于24 h和48 h进行Real-time PCR和ELISA检测细胞中IL-8 mRNA和蛋白表达。应用0.05 μg和0.2 μg pSG-GFP-LMP1质粒转染Ca9-22细胞,48 h后Real-time PCR和ELISA检测细胞中IL-8 mRNA和蛋白表达,Western blot检测p-IκBα、IκBα、p-p65和p65蛋白表达。结果: pSG-GFP-LMP1质粒转染Ca9-22细胞基因高表达,转染成功。0.2 μg质粒转染Ca9-22细胞24 h和48 h后,实验组IL-8 mRNA和蛋白表达均显著高于对照组(P<0.01)。0.05 μg和0.2 μg质粒转染Ca9-22细胞48 h后,实验组IL-8 mRNA和蛋白表达以及p-IκBα、p-p65和p65蛋白表达显著高于对照组(P<0.01),且随着剂量增大,表达增强,实验组IκBα蛋白表达显著低于对照组(P<0.01),且随着剂量增大,表达减弱。结论: EBV LMP1可能通过激活NF-κB磷酸化进而诱导Ca9-22细胞中IL-8的表达。

关键词: EB病毒潜在膜蛋白1, 白细胞介素-8, 核因子-κB, 牙龈上皮细胞

Abstract: Objective: To study the regulation of EBV LMP1 on proinflammatory factors of Ca9-22 cells as well as its mechanism. Methods: Ca9-22 cells were transfected with 0.05 μg and 0.2 μg pSG-GFP-LMP1 plasmids. The expressions of GFP were observed under fluorescence microscope, the mRNA and protein levels of LMP1 were measured by Real-time PCR and ELISA after 24 h and 48 h, respectively. Ca9-22 cells were transfected with 0.2 μg pSG-GFP-LMP1 plasmids, and the mRNA and protein levels of IL-8 were measured by real-time PCR and ELISA at 24 h and 48 h. Ca9-22 cells were transfected with 0.05 μg and 0.2 μg pSG-GFP-LMP1 plasmids, the protein levels of p-IκBα, IκBα, p-p65, and p65 were measured by western blot. Results: Ca9-22 cells transfected with the pSG-GFP-LMP1 plasmid could be expressed successfully. After transfection of Ca9-22 cells with 0.2 μg plasmid, the mRNA and protein levels of IL-8 in the experimental group were significantly increased (P<0.01). 48h after transfection, the mRNA and protein levels of IL-8, the protein levels of p-IκBα, p-p65, and p65 in the experimental group were significantly increased (P<0.01). Furthermore, the expressions were increased in a dose-dependent manner. The protein levels of IκBα was decreased in the experimental group (P<0.01) . Conclusion: EBV LMP1 may induce IL-8 expression in Ca9-22 cells by activating NF-κB phosphorylation.

Key words: Epstein-Barr virus latent membrane protein 1, IL-8, NF-κB, Ca9-22

刘芳芳, 邱忠营, 王艳丽. EBV LMP1对牙龈上皮细胞促炎因子的调控及机制研究[J]. 口腔医学研究, 2020, 36(7): 654-657.

LIU Fangfang, QIU Zhongying, WANG Yanli. Regulation and Mechanism of EBV LMP1 on Inflammatory Factors of Gingival Epithelial Cells[J]. Journal of Oral Science Research, 2020, 36(7): 654-657.

参考文献

[1] Hajishengallis G. Periodontitis: from microbial immune subversion to systemic inflammation [J]. Nat Rev Immunol, 2015, 15(1):30-44.
[2] 吴鹏,高承志.2型糖尿病患者慢性牙周炎和种植体周围炎的炎症基因表达[J].口腔医学研究,2019,35(9):854-857.
[3] 孔晨,李永丽,李思佳,等.牙周炎动物模型的研究进展[J].医学综述,2019,25(17):3344-3349.
[4] Contreras A, Nowzari H, Slots J. Herpesviruses in periodontal pocket and gingival tissue specimens [J]. Oral Microbiol Immunol, 2000, 15(1):15-18.
[5] Lu H, Zhu C, Li F, et al. Putative periodontopathic bacteria and herpesviruses in pregnant women: a case-control study [J]. Sci Rep, 2016, 6:27796.
[6] Slots J, Saygun I, Sabeti M, et al. Epstein-Barr virus in oral diseases [J]. J Periodontal Res, 2006, 41(4):235-244.
[7] Saygun I, Kubar A, Özdemir A, et al. Periodontitis lesions are a source of salivary cytomegalovirus and Epstein-Barr virus [J]. J Periodontal Res, 2005, 40(2):187-191.
[8] Konstantinidis A, Sakellari D, Papa A, et al. Real-time polymerase chain reaction quantification of Epstein-Barr virus in chronic periodontitis patients [J]. J Periodontal Res, 2005, 40(4):294-298.
[9] 谢闵,朱永云,许增祥.EB病毒潜伏膜蛋白 LMP1, VEGF 和 cyclinD1在乳腺癌中的表达及其相关性[J].现代肿瘤医学,2016,24(12):1901-1905.
[10] 曾凡新,董志,傅洁民,等.中性粒细胞弹性蛋白酶——神经保护的作用新靶点[J].中国药理学通报,2006,22(7):784-787.
[11] Imai K, Kamio N, Cueno ME, et al. Role of the histone H 3 lysine 9 methyltransferase S uv39 h1 in maintaining E psteinn-B arr virus latency in B 95-8 cells [J]. FEBS J, 2014, 281(9):2148-2158.
[12] Steven NM. Epstein-Barr virus latent infection in vivo [J]. Rev Med Virol, 1997, 7(2):97-106.
[13] 张旭.EB病毒检测及其临床应用的研究进展[J].检验医学,2018,33(3):259-263.
[14] 宋立杰,杨军星,王瑶,等.LMP-1转染人胎盘源间充质干细胞前后蛋白质组学研究[J].口腔医学研究,2017,33(5):525-529.
[15] Speck SH, Strominger JL. Transcription of Epstein-Barr virus in latently infected, growth-transformed lymphocytes [J]. Adv Viral Oncol, 1989, 8:133-150.
[16] Kato A, Imai K, Ochiai K, et al. Higher prevalence of Epstein-Barr virus DNA in deeper periodontal pockets of chronic periodontitis in Japanese patients [J]. PLoS One, 2013, 8(8):e71990.
[17] Kato A, Imai K, Ochiai K, et al. Prevalence and quantitative analysis of Epstein-Barr virus DNA and Porphyromonas gingivalis associated with Japanese chronic periodontitis patients [J]. Clin Oral Investig, 2015, 19(7):1605-1610.
[18] 王勤涛.牙周病与全身系统性疾病间的相互关系[J].国外医学:口腔医学分册,2003,30(2):135-137.
[19] Slots J. Herpesviral-bacterial interactions in periodontal diseases [J]. Periodontology 2000, 2010,52(1):117-140.
[20] Payne JB, Reinhardt RA, Masada MP, et al. Gingival crevicular fluid IL-8: correlation with local IL-1β levels and patient estrogen status [J]. J Periodontal Res, 1993, 28(6):451-453.
[21] Jin L, Söder B, Corbet EF. Interleukin-8 and granulocyte elastase in gingival crevicular fluid in relation to periodontopathogens in untreated adult periodontitis [J]. J Periodontol, 2000, 71(6):929-939.
[22] 李欣,王笑峰,刘霞,等.EB 病毒潜伏膜蛋白 LMP1 编码基因沉默对NF-κB表达的影响[J].现代生物医学进展,2009,9(6):1040-1043.
[23] 王锦华,张芳,李霞,等.NF-κB非经典途径IKKα调控的Maspin在人牙周膜细胞中的表达[J].口腔医学研究,2017,33(10):1052-1055.

EBV LMP1对牙龈上皮细胞促炎因子的调控及机制研究

Regulation and Mechanism of EBV LMP1 on Inflammatory Factors of Gingival Epithelial Cells

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 5

编辑推荐

Metrics

[1]	张晓燕, 刘运新, 吴铁. 丹参素对牙槽骨成骨细胞细胞核因子-κB受体活化因子配体/骨保护素表达的影响研究[J]. 口腔医学研究, 2019, 35(9): 894-897.
[2]	李勇伟,邱勋定,顾明,邓旎,谢莉莉. 同源形成素样蛋白2基因对口腔鳞状细胞癌侵袭、迁移及核因子-κB信号的影响[J]. 口腔医学研究, 2019, 35(2): 137-141.
[3]	李琛, 杨雪, 潘亚萍, 徐晓宇, 佟彤, 郭艳. 牙龈卟啉单胞菌唾液酸酶基因缺失影响其感染上皮细胞后炎症相关microRNA的表达[J]. 口腔医学研究, 2017, 33(2): 132-134.
[4]	赵钰婷,孟维艳. 种植体表面对牙龈上皮组织生物学行为影响的研究进展[J]. 口腔医学研究, 2015, 31(9): 942-944.
[5]	张晓燕, 崔燎, 吴铁. 基于RANKL-RANK-OPG轴研究丹参素防治牙槽骨骨质疏松的作用[J]. 口腔医学研究, 2015, 31(6): 588-590.