Department of Animal Sciences, The Ohio State University, Columbus, OH 44691, USA.
IGEVET - Instituto de Genética Veterinaria "Ing. Fernando N. Dulout" (UNLP-CONICET LA PLATA), Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, CP 1900 La Plata, Buenos Aires, Argentina.
J Anim Sci. 2023 Jan 3;101. doi: 10.1093/jas/skad310.
The aim of this experiment was to evaluate the effect of increasing dietary omega-3 (n-3) polyunsaturated fatty acid (PUFA) supplementation on plasma and follicular fluid resolvin D1 (RvD1) concentration and the mRNA expression of genes related to RvD1 production, inflammatory response, oxidative stress, hormone receptors and production, and free fatty acid receptors in the granulosa cells of ewes. Dorset × Hampshire ewes (n = 24) aged 2 to 4 yr and with an initial body weight (BW) of 84.08 ± 13.18 kg were blocked by body condition score (BCS) and BW, and randomly assigned to 12 pens. Each pen within each block was randomly assigned to one of three treatments: 1) diet without fatty acid supplementation (control), 2) diet with 0.5% n-3 PUFA supplementation (PUFA0.5), and 3) diet with 1% n-3 PUFA supplementation (PUFA1). BW, BCS, and blood samples were obtained on day 1 and every 21 d for 3 mo. Ewes were then synchronized, superstimulated, and ovariectomized. Antral follicles were aspirated to evaluate RvD1 concentration in follicular fluid, and granulosa cells were used to determine mRNA abundance. Data were analyzed as a randomized complete block design using a mixed model (MIXED or GLIMMIX with log as a link function when data presented a nonnormal distribution). A polynomial effect of treatments was used to analyze RvD1 concentration and mRNA expression when there was no interaction. In addition, the correlation between plasma and follicular fluid RvD1 concentration was evaluated. We found no differences in BW (P = 0.28) and BCS (P = 0.29) between treatments. The concentration of RvD1 in plasma and follicular fluid linearly increased (P = 0.03) and tended to increase (P = 0.06) concomitantly to increasing PUFA supplementation. Plasma and follicular fluid RvD1 concentrations were positively correlated (r = 0.61; P < 0.01). The abundance of GPX1 and GPR32 mRNA tended to increase linearly with increasing PUFA supplementation (P = 0.06). In addition, PUFA supplementation linearly decreased and tended to decrease IL-1β and COX-2 mRNA abundance (P = 0.01 and P = 0.06, respectively). In conclusion, the correlation between plasma and follicular fluid RvD1 concentration indicates a relationship between both compartments. Also, the decrease of IL-1β and the increase of GPX1 mRNA abundance after PUFA supplementation could have beneficial effects on follicle development.
本实验旨在评估增加膳食 ω-3(n-3)多不饱和脂肪酸(PUFA)补充对绵羊血浆和卵泡液分辨素 D1(RvD1)浓度以及与 RvD1 产生、炎症反应、氧化应激、激素受体和产生以及游离脂肪酸受体相关的基因 mRNA 表达的影响。将 24 只 2 至 4 岁、初始体重(BW)为 84.08±13.18kg 的 Dorset×Hampshire 母羊按体况评分(BCS)和 BW 分组,然后随机分为 12 个栏。每个栏内的羊随机分配到以下三种处理之一:1)不含脂肪酸补充剂的饮食(对照),2)0.5%n-3PUFA 补充剂的饮食(PUFA0.5),和 3)1%n-3PUFA 补充剂的饮食(PUFA1)。在 3 个月的时间里,每天和每 21d 都要测量 BW、BCS 和血液样本。然后对母羊进行同步化、超刺激和卵巢切除术。抽吸卵泡以评估卵泡液中 RvD1 的浓度,并用颗粒细胞测定 mRNA 丰度。数据采用随机完全区组设计进行分析(当数据呈非正态分布时,使用混合模型(MIXED 或 GLIMMIX,以对数作为链接函数))。当不存在相互作用时,使用处理的多项式效应来分析 RvD1 浓度和 mRNA 表达。此外,还评估了血浆和卵泡液中 RvD1 浓度之间的相关性。我们发现 BW(P=0.28)和 BCS(P=0.29)在处理之间没有差异。血浆和卵泡液中 RvD1 的浓度呈线性增加(P=0.03),并随着 PUFA 补充量的增加而呈趋势性增加(P=0.06)。血浆和卵泡液中 RvD1 浓度呈正相关(r=0.61;P<0.01)。GPX1 和 GPR32 mRNA 的丰度呈线性增加趋势,与 PUFA 补充量呈正相关(P=0.06)。此外,PUFA 补充剂线性降低,IL-1β 和 COX-2mRNA 丰度呈下降趋势(P=0.01 和 P=0.06)。总之,血浆和卵泡液中 RvD1 浓度之间的相关性表明两者之间存在关系。此外,PUFA 补充后 IL-1β 的减少和 GPX1mRNA 丰度的增加可能对卵泡发育有有益的影响。