Scientia Agricultura Sinica ›› 2007, Vol. 40 ›› Issue (1): 183-189 .doi: 10.3864/j.issn.0578-1752.at-2005-6415

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Effect of CpG-DNA on mastitis induced by Staphylococcus aureus infection in Rat Model

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  1. 南京农业大学农业部动物生理生化重点开放实验室
  • Received:2005-11-17 Revised:1900-01-01 Online:2007-01-10 Published:2007-01-10

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Abstract: Abstract: The mastitis model in rat induced by Staphylococcus aureus infection was firstly established. The S. aureus suspension containing 2×105CFU/ml(SL group), 2×1012CFU/ml(SH group) and phosphate-buffered saline (CON group) 100μl/side were inoculated into the mammary glands of rats 72h after parturition respectively and all the rats were sacrificed at 24h postinfection(n=6). The histopathologic evaluations showed that the mammary gland in SL were characterized by lipid denaturation both in alveolus; the mammary gland structure in SH were seriously destructed, polymorphonuclear neutrophils (PMN) were accumulated in alveoli. TNF-αand IL-6 in mammary gland from SH elevated significantly, however that in SL showed no significant variation. The 2×1012CFU/ml of Staphylococcus aureus were chose to evaluate the effect of CpG-DNA on mastitis . 72 lactating SD rats were randomly divided into control and treatment group. The 100μl of sterile 0.01M，pH7.2 PBS(control), CpG 200μg/rat(treatment group) were injected intramuscularly into tibialis anterior of the left leg after parturition(0h) respectively. Then 100μl /rat of bacterial suspension containing 2×1012CFU/ml(both group)of Staphylococcus aureus were inoculated into the fourth (two sides) mammary gland via the teat duct 72 hours after parturition respectively. Before and after 8h,16h,24h,48h and 72h (n=6)of inoculation, all the rats euthanatized and the mammary glands were harvested. CpG-DNA could elicit significant elevated TNF-αin mammary gland before infection, 24h and 72h postinfection. CpG-DNA could induce significant increase of IL-6 in mammary gland at 16h,24h and 48h postinfection. CpG-DNA induced significant decrease of viable bacteria at 8h,16h and 72h postinfection. Semiquantitative RT-PCR revealed that CpG-DNA induced higher level of TLR-9 mRNA in mammary gland than control. The result of the study showed that CpG-DNA induced more prompt migration of PMN from blood to mammary gland at the initial stage of Staphylococcus aureus infection, stimulated the secretion of cytokines,decreased bacteria counts in mammary gland and induced higher level of TLR-9 mRNA in mammary gland. The study indicated that to some extent the CpG-DNA protected against mastitis by Staphylococcus aureus infection in rat.

Key words: mastitis, rat, model, Staphylococcus aureu

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