中性粒细胞防御素抗SARS冠状病毒研究  

A study on antiviral activity of defensins from neutrophil of human and rabbit against SARS coronavirus in vitro

作　　者：王永杰[1] 李晓萸[2] 齐名[3] 刘军权[1] 武建国[3] 李玺[1] 金义生[1] 曹志章[1] 

机构地区：[1]解放军第97医院,徐州221004 [2]军事医学科学院微生物流行病研究所 [3]南京军区南京总医院

出　　处：《解放军医学杂志》2004年第12期1079-1081,共3页Medical Journal of Chinese People's Liberation Army

基　　金：江苏省高校自然科学研究计划项目基金资助课题 (编号 0 3KJD31 0 2 37)

摘　　要：目的　检测人和兔中性粒细胞防御素的抗SARS冠状病毒活性 ,探讨防御素在SARS发生发展过程中的作用。方法　应用酸 尿素 聚丙烯酰胺凝胶洗脱电泳(CAU PAGE)从人和兔中性粒细胞中分别纯化人中性粒细胞防御素 (HNP1 3)和兔中性粒细胞防御素(RNP1 5 ) ,通过微量培养法观察不同浓度防御素对SARS冠状病毒BJ0 1株感染Vero E6细胞及病毒复制的影响。结果　用CAU PAGE分离的HNP1 3和RNP1 5经 15 %SDS PAGE电泳后在分子量 4 0 0 0左右呈现单一条带 ,HNP1 3经ELISA定量检测确认。HNP1 3和RNP1 5的抗大肠杆菌 (ATCC2 5 92 2菌株 )活性的IC50 分别在 2 5～ 5 0 μg/ml和 5～ 10 μg/ml之间 ,保持了天然的抗菌活性。在抗SARS冠状病毒实验中 ,HNP1 3和RNP1 5在 2 5～10 0 μg/ml浓度时 ,不能阻断SARS冠状病毒致细胞病变效应 (CPE) ,也不能抑制SARS冠状病毒在Vero E6细胞中的复制。结论　天然防御素HNP1 3和RNP1 5对SARS冠状病毒没有抑制和杀灭活性 。Objective To determine the anti-SARS coronavirus (SARS-CoV) activity of defensins from human and rabbit, and explore the effects of defensin during the onset and progression of SARS. Methods Defensins were purified from neutrophil of human and rabbit respectively, namely HNP (human neutrophil protein) 1-3 and RNP (rabbit neutrophil protein) 1-5, by acid-urea-polyacrylamide gel electrophoresis (CAU-PAGE). The effect of defensin on Vero-E6 cell infected by SARS CoV BJ01 strain as well as the replication of virus were observed by micro-culture technique. Rusults The HNP 1-3 and RNP 1-5 separated by CAU-PAGE presented a single band with molecular weight of approximately 4000 dalton on 15% SDS-PAGE. HNP 1-3 was identified and validated by quantitative ELISA. The activity of anti-escherichia coli (ATCC25922 strain) of HNP 1-3 and RNP 1-5 was tested by MIC determination method, and the IC50 values obtained were 25～50μg/ml and 5～10μg/ml, respectively. In the antiviral experiment, HNP 1-3 and RNP 1-5 in the concentration from 25 to 100μg/ml were not able to prohibit cytopathic effect (CPE) of SARS-CoV and the replication of virus in Vero-E6. Conclution The natural defensins (HNP 1-3 and RNP 1-5) purified from neutrophils do not play any prohibitory or eliminative effects on SARS-CoV, indicating that that human and rabbit do not have natural immunity against SARS-CoV.

关 键 词：严重急性呼吸综合征 冠状病毒 防御素 中性粒细胞 

分 类 号：R563.19[医药卫生—呼吸系统]