Department of Agriculture and Environmental Sciences, University of Udine, Udine, Italy.
Domest Anim Endocrinol. 2012 Oct;43(3):213-26. doi: 10.1016/j.domaniend.2012.03.001. Epub 2012 Apr 4.
The aim of the present research was to investigate the regulation of gene expression in ovine blood leukocytes during ACTH-induced cortisol release and the effect of dietary administration of botanicals to counteract the evoked response in polymorphonucleate cells (PMNCs). Thirty-six homogeneous Sarda sheep (age, 18±4.1 mo; BW, 38.7±1.3 kg) were allotted to six groups of six sheep each. One group was used as a negative control (Saline) and five groups were treated, every 12 h for 48 h, with 0.5 mL of ACTH agonist (250 μg/mL of tetracosactrin). Before ACTH treatment, four of the five ACTH-treated groups were separated and fed for 22 d with a basal diet supplemented with extracts from Echinacea angustifolia roots (PO+ACTH), Echinacea angustifolia flowers (EA+ACTH), Andrographis paniculata (AP+ACTH), and the bark of Larix decidua milled (LB+ACTH). Control groups (Saline and ACTH) were fed with the same basal diet without botanicals. Total RNA was extracted from blood samples collected before (T0) and after 3 h (T3) and 51 h (T51) from the first ACTH injection, and transcriptome analysis was performed using a custom oligoarray, designed from 12,194 Ovis aries UniGenes on a CombiMatrix platform. At T3, treatment with ACTH caused down-regulation of transcripts (P<0.001) involved in "response to stress" (GADD45A, GADD45B, WRNIP1, and XRCC6) and in "innate immune response" (MAPK3 and NFkBIB). At T51, treatment with ACTH caused down-regulation (P<0.001) of genes involved in "immune response" (IFNG and IL2) and up-regulation (P<0.001) of NF-κB1 and TP53. Each botanical produced a different (P<0.001) molecular signature for these genes at T3 and T51. The most active botanical in modulating transcriptome modifications in PMNCs after ACTH-induced cortisol release was Larix decidua Mill bark followed by Polinacea roots. These botanicals can be viewed as promising feed supplements in ruminants to cope with conditions associated with increased concentrations of plasma cortisol.
本研究旨在探讨绵羊血液白细胞中基因表达在 ACTH 诱导皮质醇释放过程中的调控作用，以及植物提取物作为饲料添加剂对抗多形核细胞（PMNCs）中诱导反应的效果。将 36 只同质的萨杜绵羊（年龄 18±4.1 个月；体重 38.7±1.3kg）分为 6 组，每组 6 只。一组作为阴性对照（生理盐水），其余 5 组每 12 小时用 0.5ml 的促肾上腺皮质激素激动剂（250μg/ml 促皮质素）处理 48 小时。在 ACTH 处理前，将其中 5 个 ACTH 处理组中的 4 个分开，用基础日粮补充狭叶松果菊根提取物（PO+ACTH）、狭叶松果菊花提取物（EA+ACTH）、穿心莲提取物（AP+ACTH）和落叶松磨碎树皮提取物（LB+ACTH）喂养 22 天。对照组（生理盐水和 ACTH）用相同的基础日粮喂养，不添加植物提取物。从第一次 ACTH 注射前（T0）、3 小时（T3）和 51 小时（T51）收集血液样本，提取总 RNA，使用 CombiMatrix 平台上的定制寡核苷酸芯片进行转录组分析。在 T3 时，ACTH 处理导致参与“应激反应”（GADD45A、GADD45B、WRNIP1 和 XRCC6）和“先天免疫反应”（MAPK3 和 NFkBIB）的转录本下调（P<0.001）。在 T51 时，ACTH 处理导致参与“免疫反应”（IFNG 和 IL2）的基因下调（P<0.001）和 NF-κB1 和 TP53 的上调（P<0.001）。每种植物提取物在 T3 和 T51 时对这些基因的转录组修饰产生不同的（P<0.001）分子特征。在 ACTH 诱导皮质醇释放后，落叶松 Mill 树皮对 PMNC 中转录组修饰的调节作用最为活跃，其次是 Polinacea 根。这些植物提取物可作为反刍动物有希望的饲料添加剂，以应对与血浆皮质醇浓度升高相关的情况。
