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目的 :筛选抗内皮细胞与单核细胞黏附的活性多肽。方法 :①以氧化型低密度脂蛋白 (ox LDL ,10 0mg/L)损伤的内皮细胞作为筛选的“靶”细胞 ,从XCX15肽文库 (X代表随机氨基酸 ,C为半胱氨酸 ,X15代表编码随机 15肽 )中 ,筛选能与受损内皮细胞特异性结合的多肽。②用ELISA法鉴定部分阳性克隆 ,经DNA序列测定确定其插入的氨基酸序列。③以计数法检测特异性噬菌体多肽对内皮细胞与单核细胞黏附率的影响。结果 :①从挑选鉴定的 14个克隆中 ,得到 6个阳性克隆 ,测序结果有两个克隆的外源多肽序列相同 ,4个克隆的外源多肽中含有亮氨酸 亮氨酸重复序列。②两个序列相同的克隆的噬菌体多肽 ,能明显降低内皮细胞与单核细胞的黏附率(分别降低 17.0 %和 17.6 % ,P <0 .0 1,n =4 )。结论 :从XCX15肽文库中 ,初步鉴定出 1个具有较明显地抗内皮细胞与单核细胞黏附作用的活性多肽
Objective: Screening for active peptides that adhere to endothelial cells and monocytes. Methods: ① The endothelial cells damaged by oxidized low density lipoprotein (ox LDL) (10 0 mg / L) were selected as the “target” cells. From XCX15 peptide library (X represents random amino acid, C is cysteine, X15 represents Encoding a random 15 peptide), screening for polypeptides that specifically bind to damaged endothelial cells. (2) Some positive clones were identified by ELISA and the inserted amino acid sequence was determined by DNA sequencing. ③ Counting method was used to detect the effect of specific phage polypeptide on the adhesion rate of endothelial cells to monocytes. Results: ①There were 6 positive clones out of the 14 clones selected and identified. The sequencing results showed that the two clones had the same exogenous polypeptide sequences and the leucine leucine repeats contained in the four clones. ② The two cloned phage polypeptides with the same sequence could significantly reduce the adhesion rate of endothelial cells to monocytes (17.0% and 17.6%, P <0.01, n = 4, respectively). Conclusion: From the XCX15 peptide library, a preliminary identification of an active polypeptide with more obvious anti-endothelial cell monocyte adhesion activity