噬菌体展示技术筛选大鼠佐剂性关节炎中PGE_2受体的拮抗剂  

Screening the Antagonist of PGE_2 Receptor in Adjuvant Arthritis by Phage Display

作　　者：闫东梅[1] 于乐洋[1] 张领兵[1] 伦艳妮[1] 马威[1] 杜柏榕[1] 朱迅[1] 

机构地区：[1]吉林大学基础医学院,长春130021

出　　处：《中国生物工程杂志》2007年第3期83-87,共5页China Biotechnology

摘　　要：目的:通过噬菌体技术筛选PGE2受体拮抗剂,探讨其在类风湿性关节炎(rheumatoid arthritis,RA)中的作用,为RA的治疗开辟新的途径。方法:应用噬菌体展示技术,以PGE2为靶点,对噬菌体环七肽库进行“吸附-洗脱-扩增”筛选过程,5轮筛选后,随机挑取噬菌体克隆进行亲和力鉴定,并选取亲和力高的克隆进行测序并对序列进行分析。合成环七肽后,建立佐剂性关节炎(AA)动物模型,通过分析足爪肿胀度,以及采用ELISA方法检测关节浸液中细胞因子的含量,对小肽的抗炎作用进行探讨。结果:噬菌体经过富集后,随机挑选的23个克隆中有21个阳性克隆,而在测序的8个克隆中4个具有相同的序列,其余也具有保守氨基酸。合成的环七肽能降低AA关节的肿胀度并减少关节浸液中前炎症细胞因子的含量。结论:噬菌体筛选PGE2受体拮抗剂的方法可行,并且得到的环七肽具有一定的抗炎作用。Objectives： To screen small peptides that could antagonize the PGE2 receptor in rheumatoid arthritis （RA） by phage display technology, so that a new way could be found for the treatment of RA. Methods： The T7 select phage was screened and positive clones were selected through the circulation of absorption-ablution- amplification by phage display technology with PGE2 as the targeting selective molecule. After five circulations, phage clones were randomly selected for altlnity assessment and the clones with high affinity were picked out for sequencing analysis. Following the synthesizing of T7 peptide, the animal model of adjuvant arthritis was set up to evaluate the anti-inflammatory role of the small synthesized peptides through observing the edema level of paws and detecting proinflammatory cytokine contents by ELISA in synovial exudant. Results： After phage enrichment, 21 positive clones were picked out as positive clones amongst the 23 randomly selected clones. Within the 8 sequenced clones, 4 clones had the identical sequence and the rest clones also had conservative amino residuals. The synthesized T7 peptides could relieve paw edema and decrease the content of proinflammatory cytokines in joint exudant in RA animal model. Conclusions The method of screening the antagonists of PGE2 receptor by phage display is feasible and the T7 peptides have the anti-inflammatory effect.

关 键 词：噬菌体 PGE2受体 类风湿性关节炎 

分 类 号：Q819[生物学—生物工程]