巨噬细胞来源的凋亡外囊泡对成骨细胞增殖的影响  

The effects of macrophage-The effects of macrophage-derived apoptotic vesicles on osteoblast proliferationderived apoptotic vesicles on osteoblast proliferation

作　　者：冀吉昀 张曦戈 叶庆元 王垭铮 王晓雨 葛晓彤 李蕊 王勤涛 JI Jiyun;ZHANG Xige;YE Qingyuan;WANG Yazheng;WANG Xiaoyu;GE Xiaotong;LI Rui;WANG Qintao(State Key Laboratory of Military Stomatology&National Clinical Research Center for Oral Diseases&Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture,1.Department of Periodontology,710032 Xi'an;Digital Dentistry Center,The Third Hospital Afiliated of Air Force Military Medical University,China)

机构地区：[1]军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔生物工程技术研究中心,空军军医大学第三附属医院牙周病科,西安710032 [2]军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔生物工程技术研究中心,空军军医大学第三附属医院数字化中心,西安710032

出　　处：《实用口腔医学杂志》2023年第3期291-295,共5页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:82170955)。

摘　　要：目的:探索巨噬细胞来源的凋亡外囊泡(apoEVs)对成骨细胞增殖活性的影响。方法:使用星孢菌素诱导RAW264.7细胞凋亡,差速离心法提取上清液内的细胞外囊泡,BCA蛋白定量对细胞外囊泡进行定量后,使用不同浓度(0.05~10mg/L)的apoEVs与MC3T3-E1细胞进行共培养,通过激光共聚焦观察apoEVs是否被MC3T3-E1细胞吞噬。随后分别在24、48、72h使用MTT法测定MC3T3-E1的增殖活性。使用RNA酶对apoEVs进行处理,与MC3T3-E1细胞进行共培养一段时间后,再次使用MTT法测定MC3T3-E1细胞活性。结果:成功提取出apoEVs,5mg/L以下浓度的apoEVs使得MC3T3-E1细胞增殖活性增强,而浓度10mg/L的apoEVs使得MC3T3-E1细胞增殖活性降低;各浓度的无RNA的apoEVs均使得细胞增殖活性降低。结论:巨噬细胞来源的apoEVs在低浓度时对MC3T3-E1有促增殖能力,而高浓度时有抑制增殖能力。其中apoEVs内的RNA可能在促进增殖中发挥主要作用。Objective:To investigate the effects of macrophage-derived apoptotic extracellular vesicles(apoEVs)on the proliferation of osteoblasts.Methods:The apoptosis of RAW264.7 cells was induced by staurosporine,and the extracellular vesicles in the cell culture supernatant were extracted by differential centrifugation.Affer quantification of extracellular vesicles by BCA protein quantification,apoEVs of 0.05-10 mg/L were used to co-culture with MC3T3-El cells,confocal laser scanning microscopy was used to observe the phagocytosis of the apoEVs by MC3T3-E1 cells.The proliferation activity of MC3T3-E1 cells was determined by MTT assay at 24,48 and 72 h of the treatment,respectively.The apoEVs were treated with ribonuclease,and then co-cultured with MC3T3-E1 cells for a period of time,the proliferation of the cells was measured by MTT method.Results:ApoEVs were successfully obtained,and the apo-EVs of 0.05-5 mg/L enhanced the proliferation of MC3T3-E1 cells,while that of 10 mg/L decreased the proliferation of MC3T3-E1 cells.All concentrations of RNA-free apoevs decreased cell proliferation activity.Conclusion:Macrophage derived apoEVs may promote the proliferation of osteoblasts at low concentration,but have the ability to inhibit the proliferation at high concentration.RNA in the apoEVs may play a major role in the effects.

关 键 词：巨噬细胞 凋亡外囊泡 成骨细胞 增殖 RNA 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]