| 牛乳铁蛋白对断奶仔猪抗菌肽PMAP-37 mRNA 表达的影响 |
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| 引用本文： | 王燕,汪以真,姚国佳,单体中,徐春兰.牛乳铁蛋白对断奶仔猪抗菌肽PMAP-37 mRNA 表达的影响[J].农业生物技术学报,2007,15(5):757-761. |
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| 作者姓名： | 王燕 汪以真 姚国佳 单体中 徐春兰 |
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| 作者单位： | 浙江大学饲料科学研究所,动物分子营养学教育部重点实验室,杭州,310029 |
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| 基金项目： | 国家自然科学基金;教育部跨世纪优秀人才培养计划 |
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| 摘 要： | 本研究通过体内试验（饲养试验）和体外试验探讨了牛乳铁蛋白（bLF）对抗菌肽PMAP-37 基因表达的影响。在饲养试验中，72 头36 日龄杜长嘉(♂杜洛克×♀长嘉) 断奶仔猪, 按体重相近原则随机分成3 个处理组, 每个组设3 个重复, 每个重复8 头猪(公母各半)。对照组(处理1) 饲喂不添加bLF 的饲粮, 处理2、处理3 分别饲喂添加含0.125% bLF 和0.25% bLF 的饲粮。在体外实验中，分离培养猪骨髓细胞后，添加不同浓度的bLF（10、100、1000 µg/ml）分别培养3h 和6h，每个处理组3个重复，对照组用培养液替代。根据已报道的猪抗菌肽PMAP-37 和看家基因18S rRNA基因序列, 分别设计PMAP-37 和18S rRNA 的引物, 探讨了PCR 体系中适宜的MgCl2 浓度和循环次数。以此构建一优化的半定量RT-PCR 法。以18S rRNA 为内标，研究日粮中添加不同浓度bLF 和骨髓细胞中添加不同浓度bLF 对仔猪抗菌肽PMAP-37 基因表达的影响。结果显示，与对照组相比，日粮中添加0.125% bLF和0.25% bLF组分别使PMAP-37 基因的表达提高了109.62%（P<0.01）和69.23%（P<0.05）；在骨髓细胞中分别添加10、100、1000 µg/mL bLF在3h 和6h 诱导后对PMAP-37 基因的表达有所提高，但统计结果差异不显著。以上结果表明，bLF在体内和体外对抗菌肽PMAP-37基因表达均有影响。
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| 关 键 词： | 牛乳铁蛋白 猪 抗菌肽 PMAP-37 |
| 文章编号： | 1006-1304（2007）05-0757-05 |
| 收稿时间： | 2007-01-15 |
| 修稿时间： | 2007-03-19 |
Effect of Bovine Lactoferrin on Gene Expression of Antimicrobial Peptide PMAP-37 in Weaning Piglets |
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| WANG Yan,WANG Yi-zhen,YAO Guo-jia,SHAN Ti-zhong,XU Chun-lan.Effect of Bovine Lactoferrin on Gene Expression of Antimicrobial Peptide PMAP-37 in Weaning Piglets[J].Journal of Agricultural Biotechnology,2007,15(5):757-761. |
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| Authors: | WANG Yan WANG Yi-zhen YAO Guo-jia SHAN Ti-zhong XU Chun-lan |
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| Abstract: | This experiment was conducted to study the effect bovine lactoferrin (bLF) on antimicrobial peptide PMAP-37 gene expression. In vivo, seventy-two Duroc × Landrace × TaiHu weaning piglets (♂ Duroc× ♀Landrace× TaiHu) of 36 days were divided into three treatment groups randomly by weight and each group included three repeats of eight piglets. The control (treatment 1) received the basal dietary without bLF. Treatment 2, treatment 3 received dietary contained 0.125% bLF and 0.25% bLF respectively. In vitro, we separated bone marrow cells and cultured with bLF with 10, 100, and 1000 µg/ml respectively for 3h and 6h, and each treatment repeated three times. The control wells were incubated with culture medium. The primers for PMAP-37 and 18S rRNA were designed respectively according to the reported porcine antimicrobial peptide PMAP-37 gene and housekeeping gene 18S rRNA sequences, the suitable MgCl2 concentration and cycles in PCR system were discussed. So an optimized semi-quantitative RT-PCR was constructed, 18S rRNA used as inner control, to detect effect of different concentration bLF on antimicrobial peptide PMAP-37 gene expression in diet and in bone marrow cells of pigs. The results indicated that, compared with control group, 0.125% bLF and 0.25% bLF in diet increased the gene expression of antimicrobial peptide PMAP-37 by 109.62% (P<0.01) and 69.23% (P<0.05) respectively. In bone marrow cells, 10, 100, and 1000 µg/ml bLF also increased gene expression of PMAP-37 respectively for 3h and 6h, but statistic result was not significant. These observations suggest that antimicrobial peptide PMAP-37 gene expression could be regulated by bLF. |
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| Keywords: | PMAP-37 |
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