人子宫颈粘液抗菌活性多肽的分离纯化

目的　从人子宫颈粘液分离纯化新的抗菌多肽。方法　用 5 %乙酸提取人子宫颈粘液可溶物 ,经电泳凝胶琼脂糖弥散法抗菌试验发现 ,子宫颈粘液酸溶性提取物有一条主带对大肠杆菌耐氨苄株 ML - 35 P有强抗菌活性 ,命名为 HCP。应用制备性酸性尿素聚丙烯酰胺凝胶电泳技术及洗脱电泳技术得到构成该主带的混合物 ,最后应用反相高效液相色谱技术分离纯化多肽。经 Tricine- SDS- PAGE鉴定其相对分子质量 ,并运用琼脂糖弥散法鉴定其抗菌活性。结果　从子宫颈粘液酸溶性提取物中纯化出一多肽 ,相对分子质量约为 14× 10 3,命名为 HCP- 1,具有抗耐药性大肠杆菌和绿脓杆菌的活性。结论　 HCP- 1可能是宫颈粘液天然免疫机制中的新的效应分子

Isolation and Purification of an Antibacterial Polypeptide from Human Cervical Mucus

OBJECTIVE: This study was made to isolate and purify antibacterial polypeptides from human cervical mucus. METHODS: Human cervical mucus was collected from human healthy subjects, and the acid-soluble extract was obtained by solving the mucus with 5% acetic acid in the presence of protease inhibitor. By using gel overlay technique, one protein band that was potently antibacterial against E. coli ML-35P was identified. To purify the antibacterial polypeptide, the preparative acid-urea gel electrophoresis and Reverse Phase HPLC were performed. RESULTS: A purified antibacterial polypeptide was obtained and named HCP-1. Tricine-SDS-PAGE indicated that the relative molecular mass of HCP-1 was around 14 x 10(3). Agar radial diffusion assay showed that the purified polypeptide had antibacterial activities against E. coli ML-35P and Pseudomanas aeruginosa ATCC65922. CONCLUSION: HCP-1 may be a new effector molecule of human cervical mucus.