| CpG-DNA对金黄色葡萄球菌诱导的乳腺炎大鼠的保护研究 |
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| 引用本文： | 朱于敏,苗晋锋,邹思湘,陈伟华.CpG-DNA对金黄色葡萄球菌诱导的乳腺炎大鼠的保护研究[J].中国农业科学,2007,40(1):183-189. |
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| 作者姓名： | 朱于敏 苗晋锋 邹思湘 陈伟华 |
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| 作者单位： | 南京农业大学农业部动物生理生化重点开放实验室 |
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| 摘 要： | 【目的】建立金黄色葡萄球菌（Staphylococcus aureus）感染的大鼠乳腺炎模型，并观察CpG-DNA对乳腺的保护作用。【方法】18只雌鼠分3组(n=6)，产后72h分别灌注磷酸盐缓冲液（phosphate buffer solution，PBS）（C组）、2×105CFU•ml-1（L组）和2×1012CFU•ml-1 (H组)金葡菌到第四对乳腺内，24 h处死。L组乳腺病变轻微，H组腺泡结构破坏严重，并有大量嗜中性粒细胞（polymorphonuclear neutrophils，PMN）浸润；H组乳腺组织TNF-α、IL-6水平显著上升。选择2×1012CFU•ml-1为诱发剂量观察CpG-DNA对乳腺的保护作用：72只雌鼠分成对照和试验组(n=36)，对照组产后0 h肌注PBS；试验组肌注CpG-DNA，72 h后灌注金葡菌到第四对乳腺内。分别于灌注前（定义为0 h），灌注后8、16、24、48和72 h（n=6）处死。【结果】感染初期试验组乳腺腺泡内PMN较对照组浸润迅速。试验组乳腺组织白细胞介素-6（interleukine-6，IL-6）在16、24和48 h显著高于对照组。CpG-DNA能显著提高0、24和72 h乳腺组织肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）水平。试验组8、16和72 h的乳腺组织金葡菌数显著低于对照组。CpG-DNA能显著提高乳腺组织中其特异性受体TLR-9（toll-like receptor-9）mRNA表达水平。【结论】CpG-DNA对金葡菌感染诱发大鼠乳腺炎的乳腺有保护作用。
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| 关 键 词： | 乳腺炎 大鼠 模型 金黄色葡萄球菌 |
| 收稿时间： | 2005-11-17 |
| 修稿时间： | 2005-11-27 |
Protective Effect of CpG-DNA on Rat with Mastitis Induced by Staphylococcus aureus Infection |
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| ZHU Yu-min,Miao Jin-feng,ZOU Si-xiang,CHEN Wei-hua.Protective Effect of CpG-DNA on Rat with Mastitis Induced by Staphylococcus aureus Infection[J].Scientia Agricultura Sinica,2007,40(1):183-189. |
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| Authors: | ZHU Yu-min Miao Jin-feng ZOU Si-xiang CHEN Wei-hua |
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| Institution: | Key Laboratory Animal Physiology and Biochemistry,Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095 |
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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. |
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| Keywords: | Mastitis Rat Model Staphylococcus aureus |
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