粪肠球菌脂磷壁酸作用的炎症微环境下磷脂酶 Cγ表达研究

doi:10.13701/j.cnki.kqyxyj.2019.11.018

口腔医学研究 ›› 2019, Vol. 35 ›› Issue (11): 1084-1088.DOI: 10.13701/j.cnki.kqyxyj.2019.11.018

粪肠球菌脂磷壁酸作用的炎症微环境下磷脂酶 Cγ表达研究

史东梅, 徐慧君^#, 左美娜, 仉红, 张咪, 吴赛璇, 董明, 牛卫东^*

大连医科大学口腔医学院辽宁大连 116041

收稿日期:2019-03-02 出版日期:2019-11-28 发布日期:2019-11-21
通讯作者: 牛卫东,E-mail:nuiweidonghai@139.com
作者简介:史东梅(1990~ ),女,山东人,硕士在读,主要从事牙体牙髓病学研究。徐慧君(1994~ ),女,安徽人,硕士在读,主要从事牙体牙髓病学研究。
#为共同第一作者
基金资助:
国家自然科学基金(编号:81171538)

Study on Expression of Phospholipase Cγ in Inflammatory Microenvironment of Enterococcus Faecalis Lipoteichoic Acid

SHI Dongmei, XU Huijun^#, ZUO Meina, ZHANG Hong, ZHANG Mi, WU Saixuan, DONG Ming, NIU Weidong^*

School of Stomatology of Dalian Medical University, Dalian 116041, China

Received:2019-03-02 Online:2019-11-28 Published:2019-11-21

摘要/Abstract

摘要： 目的: 研究在粪肠球菌脂磷壁酸(lipoteichoic acid,LTA)作用的炎症微环境下,成骨和破骨细胞中磷脂酶Cγ(phospholipase c-gamma, PLCγ)和活化T细胞核因子1(nuclear factor-activated T cell 1, NFATC1)mRNA的表达。方法: 将小鼠RAW264.7及MC3T3-E1分别诱导为破骨细胞和成骨细胞。免疫荧光观察LTA作用破骨细胞及成骨细胞后PLCγ和NFATC1的定位。实时荧光定量聚合酶链反应(quantitative Real-time PCR )检测LTA作用破骨细胞和成骨细胞后PLCγ和NFATC1 mRNA的表达。细胞增殖及毒性检测试剂盒(cell counting kit-8,CCK-8)检测LTA作用破骨细胞和成骨细胞后的增殖活性。结果: 破骨和成骨细胞诱导成功。免疫荧光检测可见PLCγ定位在胞质,NFATC1定位在胞核;Real-time PCR结果发现LTA刺激后成骨细胞中PLCγ和NFATC1 mRNA表达量明显减少(P<0.05);LTA刺激后破骨细胞中PLCγ和NFATC1 mRNA表达量明显增高(P<0.05);10 mg/L LTA组可抑制成骨细胞增殖,促进破骨细胞增殖(P<0.05)。结论: PLCγ和NFATC1可能在难治性根尖周炎的炎症反应和骨破坏中发挥一定的作用。

关键词: 破骨细胞, 成骨细胞, 脂磷壁酸, 粪肠球菌, 磷脂酶Cγ

Abstract: Objective: To explore the expression of PLCγ and NFATC1 mRNA in osteoblasts and osteoclasts under the inflammatory microenvironment of Enterococcus faecalis LTA. Methods: Mouse RAW264.7 and MC3T3-E1 were induced into osteoclasts and osteoblasts, respectively. Immunofluorescence was used to observe the localization of PLCγ and NFATC1 after LTA treatment of osteoclasts and osteoblasts. Real-time PCR was used to detect the expression of PLCγ and NFATC1 mRNA after osteoclast and osteoblasts by LTA. CCK8 detects the proliferative activity of LTA after osteoclast and osteoblasts. Results: Osteogenic and osteoblasts were successfully induced. Immunofluorescence showed PLCγ localization in the cytoplasm and NFATC1 localization in the nucleus. PCR results showed that the mRNA expression levels of PLCγ and NFATC1 in osteoblasts were significantly decreased after LTA stimulation (P<0.05). The mRNA expression levels of PLCγ and NFATC1 in osteoclasts were significantly increased after LTA stimulation (P<0.05). Conclusion: PLCγ and NFATC1 may play a role in the inflammatory response and bone destruction of refractory apical periodontitis.

Key words: osteoclasts, osteoblasts, lipoteichoic acid, enterococcus faecalis, phospholipaseCγ

史东梅, 徐慧君, 左美娜, 仉红, 张咪, 吴赛璇, 董明, 牛卫东. 粪肠球菌脂磷壁酸作用的炎症微环境下磷脂酶 Cγ表达研究[J]. 口腔医学研究, 2019, 35(11): 1084-1088.

SHI Dongmei, XU Huijun, ZUO Meina, ZHANG Hong, ZHANG Mi, WU Saixuan, DONG Ming, NIU Weidong. Study on Expression of Phospholipase Cγ in Inflammatory Microenvironment of Enterococcus Faecalis Lipoteichoic Acid[J]. Journal of Oral Science Research, 2019, 35(11): 1084-1088.

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粪肠球菌脂磷壁酸作用的炎症微环境下磷脂酶 Cγ表达研究

Study on Expression of Phospholipase Cγ in Inflammatory Microenvironment of Enterococcus Faecalis Lipoteichoic Acid

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