《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (6): 1841-1848.doi: 10.16431/j.cnki.1671-7236.2019.06.033

• 基础兽医 • 上一篇    下一篇

大肠杆菌与金黄色葡萄球菌对奶牛子宫内膜组织细胞因子表达及损伤程度的影响

刘昆1, 毛伟1, 李婷婷1, 刘博1, 关红1, 裴乐2, 曹金山1   

  1. 1. 内蒙古农业大学兽医学院, 呼和浩特 010018;
    2. 内蒙古农牧业科学院, 呼和浩特 010031
  • 收稿日期:2018-09-14 出版日期:2019-06-20 发布日期:2019-06-19
  • 通讯作者: 曹金山 E-mail:jinshancao@imau.edu.cn
  • 作者简介:刘昆(1989-),男,内蒙古锡林郭勒人,博士生,研究方向:基础兽医学,E-mail:8066340@qq.com
  • 基金资助:

    国家自然科学基金(31672603)

Effect of Escherichia coli and Staphylococcus aureus on Expression of Cytokine and Damage Degree of Dairy Cows Endometrium Tissue

LIU Kun1, MAO Wei1, LI Tingting1, LIU Bo1, GUAN Hong1, PEI Le2, CAO Jinshan1   

  1. 1. College of Veterinary Medicine, Inner Mongolian Agricultural University, Hohhot 010018, China;
    2. Inner Mongolian Academy of Agriculture and Animal Husbandry Sciences, Hohhot 010031, China
  • Received:2018-09-14 Online:2019-06-20 Published:2019-06-19

摘要:

本试验旨在研究体外感染金黄色葡萄球菌与大肠杆菌对奶牛子宫内膜组织中细胞因子白介素-6(IL-6)、IL-1β和IL-8的表达及损伤程度的影响。以体外培养的奶牛子宫内膜组织作为研究对象,采用1×105~1×109 CFU/mL大肠杆菌与金黄色葡萄球菌对奶牛子宫内膜组织进行体外感染,通过实时荧光定量PCR和ELISA方法检测两种细菌刺激后奶牛子宫内膜组织中IL-6、IL-1β及IL-8 mRNA与蛋白的表达量,并用HE染色法观察两种细菌感染后奶牛子宫内膜组织病理学切片。结果显示,1×105~1×109 CFU/mL大肠杆菌体外感染后,奶牛子宫内膜组织中IL-6、IL-1β和IL-8 mRNA表达量均极显著高于空白对照组(P<0.01);金黄色葡萄球菌感染浓度为1×105、1×106 CFU/mL时,IL-6、IL-1β mRNA表达量极显著高于空白对照组(P<0.01),感染浓度为1×107 CFU/mL时,IL-6、IL-1β mRNA表达量显著高于空白对照组(P<0.05),感染浓度为1×106 CFU/mL时,IL-8 mRNA表达量显著高于空白对照组(P<0.05),其他感染浓度均与空白对照组无显著差异(P>0.05)。奶牛子宫内膜组织感染1×105~1×109 CFU/mL金黄色葡萄球菌和大肠杆菌时,IL-6、IL-1β及IL-8蛋白表达量均极显著高于空白对照组(P<0.01)。相同浓度的大肠杆菌感染奶牛子宫内膜组织后,IL-6、IL-1β及IL-8 mRNA与蛋白表达量均极显著高于金黄色葡萄球菌感染组(P<0.01)。HE切片染色结果显示,大肠杆菌感染后仍有部分上皮细胞保留,而金黄色葡萄球菌感染后上皮细胞全部脱落。本试验结果表明,大肠杆菌与金黄色葡萄球菌感染奶牛子宫内膜组织后,引起的炎症反应不同。大肠杆菌感染后,促炎性细胞因子被显著上调,而金黄色葡萄球菌感染后破坏子宫内膜上皮细胞程度更加严重。

关键词: 奶牛子宫内膜组织; 大肠杆菌; 金黄色葡萄球菌; 白介素6(IL-6); 白介素1β (IL-1β ); 白介素8(IL-8)

Abstract:

The aim of this study was to investigate the effects of Staphylococcus aureus (S.aureus) and Escherichia coli (E.coli) on the expression of cytokines interleukin-6 (IL-6),IL-1β and IL-8 and damage of dairy cow endometrial tissues.The in vitro cultured dairy cow endometrium tissue was used as the research object,and was infected in vitro with E.coli and S.aureus at a concentration of 1×105 to 1×109 CFU/mL.Real-time PCR and ELISA were used to detect the expression of IL-6,IL-1β and IL-8 mRNA and protein in dairy cows endometrium tissue after two kinds of bacteria stimulation,and the dairy cows endometrial histopathology sections were observed by HE staining.The results showed that the expression of IL-6,IL-1β and IL-8 mRNA in dairy cows endometrium tissue was extremely significantly higher than that in the blank control group after E.coli infection at a concentration of 1×105 to 1×109 CFU/mL (P<0.01).When the concentration of S.aureus infection was 1×105 and 1×106 CFU/mL,the expression of IL-6 and IL-1β mRNA was extremely significantly higher than that of the blank control group (P<0.01);The infection concentration was 1×107 CFU/mL,the expression of IL-6 and IL-1β mRNA was significantly higher than that in the blank control group (P<0.05);The infection concentration was 1×106 CFU/mL,the expression of IL-8 mRNA was significantly higher than that in the blank control group (P<0.05);Other infection concentrations were not significantly different from the blank control group (P>0.05).The expression levels of IL-6,IL-1β and IL-8 were significantly higher in the dairy cows endometrial tissues infection with 1×105 to 1×109 CFU/mL S.aureus and E.coli (P<0.01).The expression of IL-6,IL-1β and IL-8 mRNA and protein in the dairy cows endometrial tissues infected with E.coli at the same concentration was extremely significantly higher than those in the S.aureus infection group (P<0.01).The results of HE section staining showed that some epithelial cells remained after E.coli infection,while epithelial cells all fell off after S.aureus infection.The results of this test showed that the inflammatory response caused by E.coli and S.aureus infected with endometrial tissue of cows was different.After E.coli infection,pro-inflammatory cytokines were significantly up-regulated,while S.aureus infection destroyed endometrial epithelial cells to a greater extent.

Key words: dairy cows endometrium tissue; E.coli; S.aureus; IL-6; IL-1β; IL

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