Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan.
Poult Sci. 2011 Sep;90(9):2054-62. doi: 10.3382/ps.2011-01394.
The aim of this study was to determine whether the expression of proinflammatory cytokines and chemokines in ovarian cells was changed to recruit heterophils and T cells in response to lipopolysaccharide (LPS), a gram-negative bacterial component. White Leghorn laying hens were intravenously injected with LPS or saline and their ovarian follicles and stroma were collected. Changes in the mRNA expression of IL-1β, IL-6, and CXCLi2 in the theca and granulosa layers and ovarian stroma were analyzed by quantitative reverse transcriptase PCR, whereas proteins of IL-1β and IL-6 were also identified by Western blot analysis. Localization of heterophil-like cells and CD4(+) and CD8(+) T cells was examined by general histology and immunohistochemistry. The expressions of IL-1β, IL-6, and CXCLi2 were significantly increased in the granulosa layer, theca layer, and stroma tissues by 3 to 6 h after LPS injection. Increase of IL-1β and IL-6 proteins in those tissues was also identified 12 h after LPS injection. The LPS stimulation resulted in the increased influx of heterophil-like cells and CD4(+) T cells, but not CD8(+) cells, in the theca layers of follicles. Saline injection affected neither expression of examined proinflammatory cytokines and chemokines nor frequencies of immunocompetent cells. These results suggest that ovarian follicular cells and stromal cells have the ability to express proinflammatory cytokines and chemokines, and their expression is upregulated by LPS in association with the recruitment of heterophil-like cells and T cells. These responses may play roles in local host defense in ovarian follicles.
本研究旨在确定卵巢细胞中促炎细胞因子和趋化因子的表达是否发生改变，以募集异嗜粒细胞和 T 细胞来应对脂多糖（LPS），这是革兰氏阴性菌的一个组成部分。将白来航产蛋母鸡静脉内注射 LPS 或生理盐水，并收集其卵巢卵泡和基质。通过定量逆转录 PCR 分析 IL-1β、IL-6 和 CXCLi2 在卵巢的颗粒层和膜层以及基质中的 mRNA 表达变化，而 IL-1β 和 IL-6 蛋白也通过 Western blot 分析进行鉴定。通过一般组织学和免疫组织化学检查异嗜粒细胞样细胞和 CD4(+)和 CD8(+)T 细胞的定位。LPS 注射后 3 至 6 小时，颗粒层、膜层和基质组织中 IL-1β、IL-6 和 CXCLi2 的表达明显增加。LPS 注射 12 小时后，还鉴定出这些组织中 IL-1β 和 IL-6 蛋白的增加。LPS 刺激导致卵泡膜层中异嗜粒细胞样细胞和 CD4(+)T 细胞而非 CD8(+)细胞的流入增加。生理盐水注射既不影响检查的促炎细胞因子和趋化因子的表达，也不影响免疫活性细胞的频率。这些结果表明，卵巢卵泡细胞和基质细胞具有表达促炎细胞因子和趋化因子的能力，其表达可被 LPS 上调，同时募集异嗜粒细胞样细胞和 T 细胞。这些反应可能在卵巢卵泡的局部宿主防御中发挥作用。
