【摘要】 目的 :探索用电泳技术分离纯化腮腺唾液富组蛋白 (HRPs) ,并观察其抗菌活性。方法 :采用制备性酸性尿素聚丙烯酰胺凝胶电泳 (AU - PAGE) ,从健康成人刺激性腮腺唾液中分离纯化 3种主要富组蛋白 (HRP- 1、HRP- 3、HRP- 5 ) ;通过琼脂糖弥散抗菌实验 ,观察它们对变形链球菌、金黄色葡萄球菌、大肠杆菌、绿脓杆菌的抗菌活性。结果 :分离纯化出的 3种主要 HRPs经 AU - PAGE及 Tricine- SDS- PAGE分析 ,均为单一的蛋白区带 ,分子量 3～ 5k D。 3种主要 HRPs对变形链球菌和金黄色葡萄球菌均有抗菌活性 ;HRP- 3和 HRP- 5对大肠杆菌和绿脓杆菌有明显抗菌活性。结论 :采用制备性 AU - PAGE来分离 HRPs,具有分辨率高、操作简便、纯化效果好等特点。 HRPs为一组内源性广谱抗菌活性多肽 ,对革兰氏阳性和革兰氏阴性菌 ,包括一些抗生素耐药菌株 ,都有抗菌作用 ,可能是口腔抗微生物感染的重要分子 ,与口腔乃至全身健康密切相关。更多还原

【Abstract】 Objective:To isolate and purify salivary histidine rich polypeptides (HRPs) and observe their antibacterial activities.Methods: By using of preparative acid urea polyacrylamide gel electrophoresis (AU PAGE), three major HRPs(HRP 1, HRP 3 and HRP 5)were purified from parotid saliva collected from healthy adults. Their antibacterial activities were tested in the ultrasensitive antibacterial assay, named radial diffusion assay.Results: Each fraction of the three purified HRPs appeared as a single protein zone both in AU PAGE and Tricine SDS PAGE. Their molecular weights are 3~5 kD as determined by Tricine SDS PAGE. All the three major HRPs showed potent bactericidal effects on Streptococcus mutans MT6R and Staphylococcus aureas ATCC25923. As to Escherichia coli ML 35P and Pseudomonas aeruginosa ATCC27853, HRP 3 and HRP 5 also demonstrated significant antibacterial activities, whereas HRP 1 did not show any inhibitory effects on the viability of them.Conclusion: The above results suggested that the method of preparative AU PAGE we used in this study has the benefits of high resolution, simple manipulation and perfect results of purification. HRPs are important components to prevent oral tissue from bacterial infections, therefore play a potent role in the maintenance of the oral health.更多还原