人内源性抗菌肽的多样性研究  

Diversity of human endogenous antimicrobial polypeptides

作　　者：黄宁[1] 张舒[1] 杨华蓉[1] 王莉莉[1] 张旗[1] 冯云[1] 吴琦[1] 王伯瑶[1] 

机构地区：[1]四川大学华西医学中心,成都610041

出　　处：《中国抗生素杂志》2004年第5期268-271,共4页Chinese Journal of Antibiotics

基　　金：国家自然科学基金(3980 0 1 4 6);中华纽约医学基金(CMB98- 681 )

摘　　要：目的　从人单个核白细胞和子宫颈及子宫内膜粘液酸溶性提取物中分离纯化新的抗菌肽。方法　电泳凝胶琼脂糖弥散法分析 r IL- 2刺激培养的人单个核白细胞和子宫颈及子宫内膜粘液酸溶性提取物抗菌组分 ,用制备性酸性尿素聚丙烯酰胺凝胶电泳初步分离并制备电泳纯的各抗菌组分 ,反相高压液相色谱技术进一步纯化 ,琼脂糖弥散法分析其抗菌活性 ,Tricine- SDS- PAGE鉴定其纯度和分子量 ,用 Edman降解法进行氨基酸序列测定。结果　从刺激培养的人单个核白细胞分别纯化出 HL P- 2 / 2 3、HL P- 3/ 2 1两个抗菌肽分子 ,分子量分别是 7.84和 16 .86 ku;从子宫颈粘液及子宫内膜粘液分别分离纯化出 HCP- 1/ 2 1、HUP1/ 38两个抗菌肽分子 ,分子量分别为 13.89和 5 .95 ku。测定 HL P- 3/ 2 1分子的前 10位 N端氨基酸序列为PKRKAEGDAK。结论　人体抗菌肽分子存在多样性 ,HL P- 2 / 2 3、HL P- 3/ 2 1、HCP- 1/ 2 1和 HU P1/Objective To isolate and identify the new antimicrobial polypeptides from the human mononuclear leukocytes, cervix and uterus mucus. Methods Antimicrobial components of the acid soluble extracts from mononuclear leukocytes stimulated with rIL 2, cervix and uterus mucus were identified by gel over lay technique. The major antimicrobial components were isolated by preparative acid urea polyacrylamide gel electrophoresis, and they were further purified by RP HPLC. Using agarose diffusion assay, the antimicrobial activity of RP HPLC fractions was tested, and their purity and molecular mass were determined by Tricine SDS PAGE. N terminal amino acid sequence were analyzed by Edman′s reaction. Result The molecular mass of purified antimicrobial polypeptides from mononuclear leukocytes (named HLP 2/23 and HLP 3/21) and form cervix, uterus mucus (named HCP 1/21 and HUP1/38) were around 7 84ku, 16 86ku, 13 89ku, and 5 95ku respectively. The N terminal amino acid sequence of HLP 3/21 aligned as PKRKAGEDAK. Conclusion Results suggested that human endogenous antimicrobial polypeptide existed diversities. HLP 2/23, HLP 3/21, HCP 1/21 and HUP1/38 would be the new antimicrobial peptides.

关 键 词：人单个核白细胞 子宫颈 子宫内膜 内源性抗菌肽 分离纯化 抗生素 

分 类 号：R978.1[医药卫生—药品] Q786[医药卫生—药学]