磨损微粒诱导的成骨细胞自噬对肿瘤坏死因子-α及白细胞介素-6分泌的影响  

Effect of osteoblast autophagy induced by wear particles on secretion of inflammatory cytokines TNF-α and IL-6

作　　者：薛凯文 柏茂盛 赵建宁 XUE Kai-wen;BAI Mao-shen;ZHAO Jian-ning(Department of Orthopedics,Jinling Hospital,Nanjing University School of Medicine/Nanjing General Hospital of Nanjing Military Region,PLA,Nanjing 210002,Jiangsu,China)

机构地区：[1]南京大学医学院附属金陵医院(南京军区南京总医院)骨科,南京医学博士研究生210002

出　　处：《医学研究生学报》2018年第4期368-372,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81772318;81572111);江苏省自然科学基金(BK20161385)

摘　　要：目的磨损微粒诱导的成骨细胞自噬与其分泌炎症因子的关系尚未见报道。文中旨在研究无菌性松动中磨损微粒刺激成骨细胞自噬对其分泌肿瘤坏死因子α(TNF-α)及白细胞介素(IL-6)产生的影响。方法取对数生长期的小鼠成骨细胞系MC3T3-E1细胞分为3组:对照组(在α-MEM培养基培养),Co Cr Mo组(在α-MEM培养基中加入100μg/m L Co Cr Mo颗粒),3-MA组(在α-MEM培养基中加入10 mmol/L自噬抑制剂3-MA预刺激成骨细胞,3 h后加入100μg/m L Co Cr Mo颗粒),用Western blot法检测成骨细胞内自噬标志蛋白LC-3表达水平,并通过ELISA方法检测TNF-α及IL-6的分泌水平。结果与对照组和3-MA组相比,Co Cr Mo组自噬标志蛋白LC-3表达水平明显升高(P<0.05)。与对照组的TNF-α[(0.95±0.00)pg/m L]和IL-6[(4.67±0.23)pg/m L]相比,Co Cr Mo组[(10.35±3.63)、(14.27±1.07)pg/m L]、3-MA组[(302.61±45.63)、(216.67±27.71)pg/m L]分泌水平显著升高(P<0.05);其中3-MA组较Co Cr Mo组亦明显上升(P<0.01)。结论 Co Cr Mo微粒诱导的成骨细胞自噬能抑制成骨细胞中炎症因子TNF-α及IL-6分泌。Objective The relationship between osteoblast autophagy induced by wear particles and secretion of inflammatory cytokines has not been illuminated.The purpose of this study was to investigate the effect of wear particles on the secretion of TNF-αand IL-6 by autophagy of osteoblasts in aseptic loosening. Methods The mouse MC3T3-E1 osteoblasts in logarithmic growth phase were divided into 5 groups:control group(inα-MEM medium),CoCrMo particle group(100μg/mL CoCrMo particles was added inα-MEM medium),3-MA group(10 mmol/L autophagy inhibitor 3-MA prestimulated osteoblasts inα-MEM medium,3h later 100μg/mL CoCrMo particles were added),the expression of autophagy marker protein LC-3 in osteoblasts was detected by Western blotting,and the levels of TNF-αand IL-6 were detected by ELISA.Results Compared with the control group,the expression level of autophagy marker protein LC-3 was significantly increased in CoCrMo particles group(P<0.05).Compared with CoCrMo particles group,the expression level of LC-3 in 3-MA group was significantly decreased(P<0.05).Compared with control group TNF-α[(0.95±0.00)pg/mL]and IL-6[(4.67±0.23)pg/mL],CoCrMo particles group TNF-α[(10.35±3.63)pg/mL]and IL-6[(14.27±1.07)pg/mL]secretion level was significantly increased(P<0.05),compared with CoCrMo particles group,the effects of 10 mmol/L 3-MA group to TNF-α[(13.13±5.08)(14.71±5.46)pg/mL]and IL-6[(15.31±0.88),(16.75±3.87)pg/mL]secretion were not significant,while 10 mmol/L 3-MA group TNF-α[The secretion levels of(302.61±45.63)pg/mL]and IL-6[(216.67±27.71)pg/mL]increased significantly(P<0.01).Conclusion [WTBZ]The osteoblast autophagy induced by CoCrMo particles can inhibit the secretion of inflammatory cytokines TNF-αand IL-6 in osteoblasts.

关 键 词：磨损微粒 自噬 TNF-α IL-6 

分 类 号：R687.4[医药卫生—骨科学]