Zhu Yu-Min, Miao Jin-Feng, Zhang Yuan-Shu, Li Zhen, Zou Si-Xiang, Deng Yue-E
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
Vet Immunol Immunopathol. 2007 Dec 15;120(3-4):168-76. doi: 10.1016/j.vetimm.2007.08.003. Epub 2007 Aug 10.
Seven healthy native goats in early lactation, weighing 30-40 kg, were used in this study. The right mammary gland of the seven does were infused with CpG-ODN at a dosage of 100 microg kg(-1) body weight on the day 5 postpartum (PP). The left glands were used as controls and infused with sterile phosphate-buffered saline (PBS). On day 8 PP, the same dosage of CpG-ODN or PBS was again infused. On day 9 PP, the mammary glands (both right and left) of the seven does were infused with 6 x 10(6) colony-forming units (CFU) Escherichia coli and, at 0, 8, 16, 24, 48 and 72 h postinfection (PI), milk samples were collected from all glands. Goats were euthanized at 72 h PI and the mammary tissue harvested. Infusion with 6 x 10(6)CFU ml(-1)E. coli induced acute mastitis. Histopathological evaluations showed that polymorphonuclear neutrophils (PMNs) were still present in alveoli at 72 h PI, but PMNs in the CpG-ODN-treated glands has disappeared. Bacteria counts in milk peaked at 16 h PI and CpG-ODN induced a significant decrease in viable bacteria from 16 h PI until the end of the experiment. This study showed that CpG-ODN promoted the expression of its specific receptor (TLR-9 mRNA) in mammary tissue, stimulated IL-6 production, reduced bacteria counts in milk, attenuated the impact of inflammation mediators on cells and significantly shortened the inflammation course. These results suggest that the CpG-ODN improved mammary gland defense and, thereby, had a beneficial effects against mastitis caused by E. coli infection in goats.
本研究选用7只处于泌乳早期、体重30 - 40千克的健康本地山羊。在产后第5天,给这7只母羊的右侧乳腺注入剂量为100微克/千克体重的CpG-ODN。左侧乳腺作为对照,注入无菌磷酸盐缓冲盐水(PBS)。产后第8天,再次注入相同剂量的CpG-ODN或PBS。产后第9天,给7只母羊的双侧乳腺注入6×10⁶ 菌落形成单位(CFU)的大肠杆菌,在感染后0、8、16、24、48和72小时,从所有乳腺采集乳汁样本。在感染后72小时对山羊实施安乐死并采集乳腺组织。注入6×10⁶CFU/毫升的大肠杆菌诱发了急性乳腺炎。组织病理学评估显示,在感染后72小时,多形核中性粒细胞(PMN)仍存在于肺泡中,但经CpG-ODN处理的腺体中的PMN已消失。乳汁中的细菌计数在感染后16小时达到峰值,CpG-ODN导致从感染后16小时直至实验结束时活菌数量显著减少。本研究表明,CpG-ODN促进其特异性受体(TLR-9 mRNA)在乳腺组织中的表达,刺激白细胞介素-6的产生减少乳汁中的细菌数量,减轻炎症介质对细胞的影响,并显著缩短炎症进程。这些结果表明,CpG-ODN改善了乳腺防御功能,从而对山羊大肠杆菌感染引起的乳腺炎具有有益作用。