Institució Catalana de Recerca i Estudis Avançats (ICREA), 08010 Barcelona, Spain; Department of Ruminant Production, Institut de Recerca i Tecnologia Agroalimentàries (IRTA), 08140 Caldes de Montbui, Spain.
Blanca from the Pyrenees, 25795 Hostalets de Tost, Spain.
J Dairy Sci. 2018 Mar;101(3):2631-2640. doi: 10.3168/jds.2017-13212. Epub 2017 Dec 28.
The objectives of this study were (1) to use endoscopy to collect biopsies from the rumen and colon epithelia to describe changes in gene expression in these 2 tissues as cows move from a dry to a lactation ration and (2) to evaluate the potential influence that supplementation of live yeast could exert on these 2 epithelia. Twenty-one Holstein cows were split into 2 treatments and received either 300 g/d of corn containing 1 × 10 cfu/d of live yeast (LY; n = 10) or 300 g/d of corn with no supplementation (control; n = 11) starting 21 ± 2.6 d (average ± SD) before until 21 d after calving. At 14 ± 2.6 d before the expected calving date, and exactly at 7 and 21 d after calving, rumen and colon biopsies were obtained from each cow using an endoscope. Total RNA was extracted from rumen and colon tissues, and the expression of IL10, TNFA, TLR4, IL1B, PCNA, MKI67, SGLT1, BAX, CASP3, OCLN, CLDN4, HSPA1A, HSPB1, DEFB1, and MCT1 (the latter only in rumen samples) was quantified by quantitative PCR. Overall, fluctuations in expression of the selected genes in the colon between the 2 stages of production and the 2 treatments were smaller than those found in the rumen. In the rumen epithelium, expression of TLR4 and DEFB1 was greatest before calving, with LY cows having a greater expression of TLR4 than control cows. Similarly, expression of IL10 was greatest in LY cows before calving. Expression of TNFA in the rumen epithelium of control cows was lowest at 21 DIM but in LY cows was kept steady among production stages. The expression of PCNA and MKI67 in the rumen epithelium was greatest at 7 DIM, indicating a high proliferation rate of this epithelium after calving. In the colon mucosa, expression of TLR4 and DEFB1 was greater than in the rumen, and DEFB1 expression was greater in LY cows than in control cows. The use of an endoscope allowed us to study the dynamics of rumen epithelium adaptation to increased supply of concentrate after calving, consisting of increased epithelia remodeling, reduction of the TLR4, and increased IL10 expression. Furthermore, the rumen epithelium of dry cows responded rapidly to live yeast, with changes in the expression of genes involved in the immune response becoming evident after 7 d of exposure to yeast. The expression of genes related to the immune response (mainly TLR4 and DEFB1) in the colon mucosa was greater than in the rumen, and the expression of DEFB1 was further stimulated by live yeast. It is concluded that the use of an endoscope allows the study of gene expression patterns in the rumen and hindgut epithelia. We report marked changes in the rumen wall and more modest changes in the colon when transitioning from a dry to a lactation ration. Furthermore, supplementation of live yeast fostered and increased expression of genes regulating inflammation and epithelial barrier in the rumen, and in the colon it increased the expression of DFEB1 coding for an antimicrobial peptide.
(1) 使用内窥镜从瘤胃和结肠上皮采集活检样本,描述奶牛从干奶期到泌乳期时这 2 种组织中基因表达的变化;(2) 评估添加活酵母对这 2 种上皮组织的潜在影响。21 头荷斯坦奶牛被分为 2 个处理组,分别在干奶前 21 ± 2.6 d(平均 ± SD)至产后 21 d 期间,每天每头奶牛分别给予含有 1 × 10 cfu/d 活酵母的 300 g/d 玉米(LY;n = 10)或无补充的 300 g/d 玉米(对照;n = 11)。在预计产犊日期前 14 ± 2.6 d 以及产后 7 和 21 d,使用内窥镜从每头奶牛获得瘤胃和结肠活检样本。从瘤胃和结肠组织中提取总 RNA,并通过定量 PCR 定量检测 IL10、TNFA、TLR4、IL1B、PCNA、MKI67、SGLT1、BAX、CASP3、OCLN、CLDN4、HSPA1A、HSPB1、DEFB1 和 MCT1(仅在瘤胃样本中)的表达。总的来说,与生产阶段和 2 种处理相比,结肠中所选基因的表达在这 2 个阶段之间的波动较小。在瘤胃上皮中,TLR4 和 DEFB1 的表达在产犊前最大,LY 牛的 TLR4 表达大于对照牛。同样,IL10 的表达在产犊前的 LY 牛中最大。在产犊后 21 DIM 时,对照牛的瘤胃上皮中 TNFA 的表达最低,但在 LY 牛中保持稳定。在瘤胃上皮中,PCNA 和 MKI67 的表达在 7 DIM 时最高,表明产后该上皮的增殖速度较高。在结肠黏膜中,TLR4 和 DEFB1 的表达大于瘤胃,并且 LY 牛中 DEFB1 的表达大于对照牛。使用内窥镜可以研究瘤胃上皮在产犊后增加浓缩物供应时的适应性动态,包括上皮重塑增加、TLR4 减少和 IL10 表达增加。此外,干奶牛的瘤胃上皮对活酵母的反应迅速,暴露于酵母 7 d 后,与免疫反应相关的基因表达变化变得明显。结肠黏膜中与免疫反应相关的基因(主要是 TLR4 和 DEFB1)的表达大于瘤胃,并且活酵母进一步刺激了 DEFB1 的表达。综上所述,使用内窥镜可以研究瘤胃和后肠上皮中的基因表达模式。我们报告了从干奶期到泌乳期时瘤胃壁的明显变化和结肠的变化较小。此外,活酵母的添加促进并增加了瘤胃中调节炎症和上皮屏障的基因的表达,在结肠中增加了编码抗菌肽的 DEFB1 的表达。