Eubiotics是家禽中使用的水和/或饲料添加剂,用于促进肠道健康和控制病原体的肠道负担。包括产气荚膜梭菌.虽然几种优生化合物(EC)正在商业化引入,在体内测试之前,必须设计一个体外模型来筛选这些化合物,以评估它们的免疫调节和抗微生物特性。鸡巨噬细胞系(MQ-NCSU)用于建立体外模型以筛选免疫学和抗C。10种ECs的产气荚膜特性:单丁酸甘油酯,monolaurin,丁酸钙,三丁酸甘油酯,香芹酚,姜黄素,绿茶提取物,迷迭香提取物,单核细胞,还有酒石酸.通过测量对MQ-NCSU细胞活力的影响来选择每种EC的最佳浓度。然后用EC处理细胞6、12和24小时,并表达干扰素-γ(IFNγ),白细胞介素(IL)-1β,IL-6,IL-10,转化生长因子β(TGFβ)和分化簇(CD40)基因,以及主要组织相容性复合体(MHC)-II蛋白进行了评估。在刺激后6小时,单丁酸甘油酯,丁酸钙,和绿茶提取物处理诱导IFNγ的显著下调,IL-6或IL-1β基因转录和MHC-II表达,而在这些治疗以及接受迷迭香提取物和酒石酸的治疗中,IL-10或TGFβ基因表达显着上调,与对照相比,提示这些ECs的免疫调节特性。最后,用这些选择的5个ECs预处理巨噬细胞24小时,然后是产气荚膜梭菌感染显示单丁酸甘油酯,绿茶提取物,迷迭香提取物,和丁酸钙治疗可以在感染后12和/或24小时显着抑制细菌生长,与对照组相比。总的来说,我们的发现表明,ECs具有免疫调节和抗C。可以使用基于体外禽类巨噬细胞的模型选择产气荚膜性质,使得可以进一步在体内测试此类EC的疾病预防功效。
Eubiotics are water and/or feed additives used in poultry to promote gut health and control enteric burden of pathogens, including Clostridium perfringens. While several eubiotic compounds (ECs) are being introduced commercially, it is essential to devise an in vitro model to screen these compounds to assess their immunomodulatory and antimicrobial properties prior to their testing in vivo. A chicken macrophage cell-line (MQ-NCSU) was used to develop an in vitro model to screen the immunological and anti-C. perfringens properties of 10 ECs: monobutyrin, monolaurin, calcium butyrate, tributyrin, carvacrol, curcumin, green tea extract, rosemary extract, monomyristate, and tartaric acid. An optimal concentration for each EC was selected by measuring the effect on viability of MQ-NCSU cells. Cells were then treated with ECs for 6, 12, and 24 h. and expression of interferon-gamma (IFNγ), interleukin (IL)-1β, IL-6, IL-10, transforming growth factor-beta (TGFβ) and cluster of differentiation (CD40) genes, as well as major histocompatibility complex (MHC)-II protein were evaluated. At 6 h post-stimulation, monobutyrin, calcium butyrate, and green tea extract treatments induced a significant downregulation of IFNγ, IL-6, or IL-1β gene transcription and MHC-II expression, while the IL-10 or TGFβ gene expression in these treatments as well as those receiving rosemary extract and tartaric acid was significantly upregulated, when compared to control, suggesting immunomodulatory properties of these ECs. Finally, pretreatment of macrophages with these selected 5 ECs for 24 h followed by C. perfringens infection showed that monobutyrin, green tea extract, rosemary extract, and calcium butyrate treatments can inhibit bacterial growth significantly at 12 and/or 24 h post-infection, when compared to the control. Collectively, our findings show that ECs possessing immunomodulatory and anti-C. perfringens properties can be selected using an in vitro avian macrophage cell-based model so that such ECs can further be tested in vivo for their disease prevention efficacy.