NBD多肽与OPG联合抑制聚乙烯磨损颗粒诱导的溶骨效应  

NBD peptide in combination with osteoprotegerin inhibits bone destruction induced by polyethylene wear particles in mouse air pouch model

作　　者：汪洋[1,2] 张健[3] 周锐[4] 成名翔[5] 曾理[6] 吴宁宁[2] 李锐冬[1,2] 牟钰钦[6] 邓忠良[1] 

机构地区：[1]重庆医科大学附属第二医院骨科,重庆400010 [2]重庆医科大学检验系临床检验诊断学教育部重点实验室,重庆400016 [3]重庆医科大学附属第一医院骨科,重庆400016 [4]第三军医大学大坪医院野战外科研究所创伤实验室,创伤,烧伤与复合伤国家重点实验室,重庆400042 [5]重庆医科大学附属第二医院肝胆外科,重庆400010 [6]重庆医科大学药学院,重庆400016

出　　处：《第三军医大学学报》2011年第17期1789-1793,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(81071490;30772211)~~

摘　　要：目的运用小鼠植骨气囊模型,研究NBD多肽、NBD多肽联合骨保护素(osteoprotegerin,OPG)减少骨破坏吸收防治人工关节松动的效应。方法在小鼠背部注入空气形成气囊,取同源小鼠的颅骨植入气囊内。将已制成的气囊模型的小鼠分成3组:NBD多肽组(气囊内注入聚乙烯颗粒及NBD多肽);联合组(气囊内注入聚乙烯颗粒、NBD多肽及OPG);对照组(气囊注入聚乙烯颗粒及生理盐水)。3周后取囊壁和植入颅骨组织进行HE染色检测炎症反应情况及炎症细胞渗出量;取组织匀浆液应用ELISA方法检测炎性细胞因子(IL-1、TNF)浓度;抗酒石酸酸性磷酸酶染色观察破骨细胞的分化成熟情况;应用扫描电镜检测骨片吸收情况及RT-PCR检测破骨细胞骨吸收相关基因Ⅱ型碳酸酐酶(CA-Ⅱ)mRNA表达水平。结果①HE染色检测炎症细胞渗出量NBD多肽组(870±236)、联合组(820±198)明显少于对照组(3 325±467)(P<0.05)。②ELISA法检测炎性细胞因子(IL-1、TNF)浓度NBD多肽组[(138.34±2.32)、(156.14±4.17)]与联合组[(129.32±4.62)、(148.78±5.36)]均明显低于对照组[(187.56±4.78)、(179.45±8.34),P<0.05]。③抗酒石酸酸性磷酸酶染色阳性区域与视野面积的百分比NBD多肽组[(1.06±0.67)%]与联合组[(0.64±0.33)%]明显少于对照组[(4.12±1.09)%],且联合组较NBD多肽组染色面积也有显著减少(P<0.05)。④应用扫描电镜检测骨片吸收陷窝面积与视野面积的百分比NBD多肽组[(4.03±0.76)%]与联合组[(2.65±0.58)%]明显低于对照组[(12.38±1.98)%];且联合组较NBD多肽组的骨片吸收面积有显著减少(P<0.05)。⑤破骨细胞骨吸收功能相关基因CA-ⅡmRNA相对表达量NBD多肽组(0.254±0.048)与联合组(0.180±0.033)明显少于对照组(0.382±0.072)。且与NBD多肽组相比,联合组表达水平也有显著减少(P<0.05)。结论 NBD多肽能明显减轻聚乙烯磨损颗粒所致的炎症反应,从而间接抑制破骨细胞激活,有效减少聚乙烯磨损颗�Objective To observe the effects of NF-κB essential modifier binding domain（NBD） peptide,alone or in combination with osteoprotegerin（OPG）,on bone destruction reduction as well as prevention and treatment of artificial joint loosening with a mouse air pouch model.Methods The mouse air pouch model was constructed through injecting air into mouse back to form a pouch and implanting skull bone from homologous mouse into the air pouch.The model mice were divided into three groups： an NBD peptide group（polyethylene particles and NBD peptides were injected into the air pouch）,a combination group（polyethylene particles,NBD peptides,and OPG were injected into the air pouch）,and a control group（polyethylene particles and saline were injected into the air pouch）.Pouch tissues and implants were collected after three weeks for detection.HE staining was used to detect inflammatory response and inflammatory cell infiltration.ELISA was used to detect cytokine（IL-1 and TNF） concentrations in tissue homogenate.Tartrate-resistant acid phosphatase staining was used to observe osteoclast differentiation and maturation.Scanning electron microscopy was used to detect bone slice resorption.RT-PCR was used to detect type-Ⅱ carbonic anhydrase（CA-Ⅱ） mRNA expression.Results HE staining results showed that inflammatory cell amounts in the NBD peptide group（870±236） and combination group（820±198） were significantly smaller than that in the control group（3 325±467）（P0.05）.ELISA results showed that inflammatory cytokine（IL-1 and TNF） concentrations in the NBD peptide group（138.34±2.32 and 156.14±4.17） and combination group（129.32±4.62 and 148.78±5.36） were significantly lower than those in the control group（187.56±4.78 and 179.45±8.34）（P0.05）.Tartrate-resistant acid phosphatase staining results showed that the percentages of positive area to visual field area in the NBD peptide group [（1.06±0.67）%] and combination group [（0.64±0.33）%] were significantly l

关 键 词：NBD多肽 骨保护素 人工关节 植骨气囊动物模型 无菌松动 

分 类 号：R318.08[医药卫生—生物医学工程] R322.71[医药卫生—基础医学]