抗RANKL多克隆抗体对P.gingivalis感染的大鼠牙周骨吸收的抑制作用  

The Inhibition of anti-RANKL Antibody on Periodontal Bone Resorption Caused by P.gingivalis in Rat

作　　者：林晓萍[1,2] 韩晓哲 魏巍[1] 周晓佳[1] Martin A.Taubman 

机构地区：[1]中国医科大学附属盛京医院口腔科,辽宁沈阳110004 [2]Department of Immunology,The Forsyth Institute [3]Department of Immunology The Forsyth Institute

出　　处：《解剖科学进展》2009年第3期269-272,共4页Progress of Anatomical Sciences

摘　　要：目的评价抗NF-кB受体活化因子配体(RANKL)多克隆抗体对P.gingivalis感染引起的牙周骨吸收的抑制作用。方法将大鼠重组的RANKL对兔进行免疫获得兔抗鼠RANKL多克隆抗体,应用兔抗鼠RANKLF(ab')2抗体片断以防止免疫抑制。实验用大鼠口腔连续4d感染活P.gingivalis(109/ml/d),第5、9及14天于腭侧牙龈乳头处注射兔抗鼠RANKLF(ab')2抗体片断(1μg/部位),OPG-Fc(1μg/部位)及不相关细胞因子L6-Fc(1μg/部位),第28天处死,取样待检。取实验当天、实验14天及28天血清,ELISA法测定血清抗P.gingivalis的特异性IgG抗体滴度及牙龈组织匀浆液中的可溶性RANKL(sRANKL)的表达水平,采用SPSS11.5统计软件包进行分析。结果口腔感染P.gingivalis后血清抗P.gingivalis的特异性IgG抗体滴度明显升高,且一直持续到第28天;局部注射抗RANKL抗体并不降低血清中抗P.gingivalis的特异性IgG抗体滴度。注射抗体组和OPG-Fc组的sRANKL的表达明显下降(<0.05),与对照组相比具有统计学意义,而注射L6-Fc组sRANKL的表达无改变;牙周骨吸收的水平变化与牙龈组织匀浆液中RANKL的表达水平相一致。结论抗RANKL多克隆抗体可降低牙龈组织中的可溶性RANKL的含量,抑制P.gingivalis感染的牙周骨吸收。Objective To evaluate the inhibition of anti- receptor activator of NF- κB ligand （RANKL） antibody on P. gingivalis induced periodontal bone resorption（PBR） in rat. Methods Rabbit anti-RANKL antibody was produced by immunization with recombinant rat RANKL and rabbit anti-RANKL F（ab＇）2 antibody fragments were used to prevent their immunogenic in the model of PBR. The inoculation was administered by oral gavages using live P.gingivalis （10^9/ml/day） for consecutive 4 days. Rabbit anti-RANKL F（ab＇）2 fragments （1 μg/site）, OPG-Fc （1 μg/site） and irrelevant cytokine L6-Fc（1 μg/site） were injected into the palatal gingivae of molars in each rat （Days 5, 9, or 12）. ELISA was used to determine rat serum lgG antibody to rabbit and soluble RANKL （sRANKL） levels in gingival homogenates. Periodontal bone resorption was measured on the maxillary molars, SASS 11.5 software package was used for statistical analysis. Results Rat serum special IgG antibody titer toP. gingivalis significantly increased until days 28 after oral infection, but not decreased by injection of anti-RANKL F（ab＇）2 fragments, or OPG-Fc or L6-Fc. sRANKL concentrations in the gingival homogenates of anti- RANKL antibidy-treated group and OPG-Fc-treated group decreased significantly compared to untreated group （P〈 0.05）, but no difference compared to L6-Fc-treated group. Anti-RANKL antibidy inhibited periodontal bone resorption significantly, the level of bone loss was parallel to the RANKL expression level in the gingival tissue homogenates. Conclusion Anti- RANKL antibody can inhibit P. gingivalis induced periodontal bone resorption and decrease the content of sRANKL in gingival tissue, which provides an immunological approach to ameliorate periodontal bone resorotion.

关 键 词：核因子-КB受体活化因子配体 免疫反应 牙龈卟啉单胞菌 大鼠 牙周骨吸收 

分 类 号：R781.4[医药卫生—口腔医学]