Bilby T R, Guzeloglu A, MacLaren L A, Staples C R, Thatcher W W
Department of Animal Sciences, University of Florida, Gainesville, Florida 32611-0910, USA.
J Dairy Sci. 2006 Sep;89(9):3375-85. doi: 10.3168/jds.S0022-0302(06)72374-8.
The objectives were to examine the effects of bovine somatotropin (bST), pregnancy, and dietary fatty acids on expression of key endometrial genes and proteins regulating prostaglandin synthesis in lactating dairy cows. Two diets were fed, at about 17 d in milk (DIM), in which oil of whole cottonseed (control diet) was compared with calcium salts of fish oil-enriched lipid (FO). Ovulation was synchronized in cows with a presynchronization plus Ovsynch protocol and cows were inseminated artificially or not inseminated on d 0 (d 0 = time of synchronized ovulation; 77 +/- 12 DIM). On d 0 and 11, cows received bST (500 mg) or no bST, and were slaughtered on d 17 to recover uterine secretions and endometrial tissue. Number of cows in the control diet: 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows and in the FO diet: 4 bST-treated FO-cyclic (bST-FO-C) and 5 non-bST-treated cyclic (no bST-FO-C) cows. The FO diet increased progesterone receptor (PR) mRNA, and treatment with bST increased PR mRNA concentration in endometrium of no bST-C, but not in no bST-FO-C or no bST-P cows. Concentrations of estrogen receptor-alpha (ERalpha) mRNA and protein, and oxytocin receptor (OTR) mRNA were decreased in no bST-P cows compared with no bST-C cows. Treatment with bST tended to increase OTR and ERalpha mRNA concentrations in cyclic cows fed control or FO diets. Immunohistochemistry demonstrated effects of bST, FO, and pregnancy on distributions of ERalpha and PR proteins in endometrium. Pregnancy and FO feeding decreased ERalpha abundance in luminal epithelium. Prostaglandin H synthase-2 (PGHS-2) protein was elevated in pregnant cows and localized to the luminal epithelium. Both FO and bST treatments reduced staining intensity of PGHS-2 protein. Concentrations of prostaglandin E synthase mRNA were elevated in either cyclic or pregnant cows in response to bST, whereas bST decreased prostaglandin F synthase mRNA in pregnant cows. Uterine lumen fluids had more PGF2alpha and prostaglandin E2 in pregnant than cyclic cows. Uterine lumen fluids of bST-P cows contained more prostaglandin E2 than those from no bST-P cows. In summary, both pregnancy and bST altered endometrial gene expression, and cyclic cows responded differently to bST than pregnant cows. Feeding FO modulated PR, ERalpha, and PGHS-2 expression and distribution among endometrial cell types in a manner that may favor establishment and maintenance of pregnancy.
本研究旨在探讨牛生长激素(bST)、妊娠和日粮脂肪酸对泌乳奶牛子宫内膜中调控前列腺素合成的关键基因和蛋白质表达的影响。在产奶约17天时(DIM),给奶牛饲喂两种日粮,将全棉籽(对照日粮)与富含鱼油的脂质钙盐(FO)进行比较。采用预同步加Ovsynch方案使奶牛排卵同步,在第0天(第0天 = 同步排卵时间;77±12 DIM)对奶牛进行人工授精或不授精。在第0天和第11天,给奶牛注射bST(500 mg)或不注射bST,并在第17天屠宰以收集子宫分泌物和子宫内膜组织。对照日粮组的奶牛数量为:5头接受bST处理的周期性奶牛(bST-C)、5头未接受bST处理的周期性奶牛(未注射bST-C)、4头接受bST处理的妊娠奶牛(bST-P)和5头未接受bST处理的妊娠奶牛(未注射bST-P);FO日粮组的奶牛数量为:4头接受bST处理的FO周期性奶牛(bST-FO-C)和5头未接受bST处理的周期性奶牛(未注射bST-FO-C)。FO日粮增加了孕酮受体(PR)mRNA的表达,bST处理增加了未注射bST-C组奶牛子宫内膜中PR mRNA的浓度,但未注射bST-FO-C组和未注射bST-P组奶牛未出现这种情况。与未注射bST-C组奶牛相比,未注射bST-P组奶牛中雌激素受体α(ERα)mRNA和蛋白质以及催产素受体(OTR)mRNA的浓度降低。bST处理倾向于增加饲喂对照或FO日粮的周期性奶牛中OTR和ERα mRNA的浓度。免疫组织化学显示bST、FO和妊娠对子宫内膜中ERα和PR蛋白分布的影响。妊娠和饲喂FO降低了腔上皮中ERα的丰度。前列腺素H合成酶-2(PGHS-2)蛋白在妊娠奶牛中升高,并定位于腔上皮。FO和bST处理均降低了PGHS-2蛋白的染色强度。bST处理使周期性或妊娠奶牛中前列腺素E合成酶mRNA的浓度升高,而bST降低了妊娠奶牛中前列腺素F合成酶mRNA的浓度。妊娠奶牛子宫腔液中的前列腺素F2α和前列腺素E2比周期性奶牛更多。bST-P组奶牛子宫腔液中的前列腺素E2比未注射bST-P组奶牛的更多。总之,妊娠和bST均改变了子宫内膜基因表达,周期性奶牛对bST的反应与妊娠奶牛不同。饲喂FO以可能有利于妊娠建立和维持的方式调节PR、ERα和PGHS-2在子宫内膜细胞类型中的表达和分布。