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1. Experimental background: Early pregnancy abortion is an important issue in humans and animals. The early pregnancy abortion rate in mammals is as high as 30%-50%. Arginine activates the mammalian target of rapamycin (mTOR) signaling pathway, which stimulates the synthesis of placenta, uterus and fetal proteins, and has important effects on embryo formation and the entire pregnancy process. Dietary supplementation with Arg improves embryonic survival in early pregnancy, promotes placental growth, and prevents fetal growth restriction. However, in the production practice, due to the added cost and formula, the exogenously added arginine failed to fully function in the animal body. N-carbamoylglutamic acid (NCG) can act as an endogenous arginine metabolic activator with stable physicochemical properties, does not interfere with intestinal absorption of tryptophan and other basic amino groups, and continuously activates melon in the intestine. The synthesis of acid promotes the endogenous synthesis of arginine to achieve a balanced supply. Studies have shown that NCG can increase the survival rate of early embryos in rats. However, little is known about the key factors in the addition of NCG in sow diets to increase embryo implantation rate and oxytocin in the endometrium.
2. Purpose of the experiment: The purpose of this study is to investigate the effect of adding NCG to the survival and development of embryos in the diet of sows in early pregnancy, and to reveal the analysis methods of isotopically labeled relative and absolute quantitation (iTRAQ) endometrial proteomics. Relevant key factors.
3. Test method: The test was divided into 14 groups and 28 days of sows and fetal pigs. The test was divided into two groups. The control group was fed with basal diet, and the control group was supplemented with 0.05% NCG in the basal diet. The effect of the addition of NCG on the endometrial proteome was examined using the quantitative proteomics approach of iTRAQ. The purity of NCG used in the test was ≥97% and was provided by the National Feed Engineering Technology Research Center.
4. Results:
1. The value is average ± standard error. n = 8 (control) or 7 (NCG). * indicates P < 0.05 compared with the control group, and ** indicates P < 0.01. NCG is N-carbamoylglutamic acid.
2. The control group was fed a basal diet.
3. The test group diet was supplemented with 0.05% NCG in the basal diet.
1. The value is average ± standard error. At 14 days of gestation , n = 7 (control group) or 8 (NCG); at 28 days of gestation, n = 8 (control) or 7 (NCG). * indicates P < 0.05 compared with the control group, and ** indicates P < 0.01. NCG is N-carbamoylglutamic acid.
　　 2. The control group was fed a basal diet.
3. The test group diet was supplemented with 0.05% NCG in the basal diet.
1. The value is average ± standard error. At 14 days of gestation, n = 7 (control group) or 8 (NCG); at 28 days of gestation, n = 8 (control) or 7 (NCG). * indicates P < 0.05 compared with the control group, and ** indicates P < 0.01. NCG is N-carbamoylglutamic acid.
2. The control group was fed a basal diet.
3. The test group diet was supplemented with 0.05% NCG in the basal diet.
　 4.1 The impact of NCG on sow reproductive performance
The results showed that the addition of NCG to the diet at 28 days of gestation significantly increased the total litter size of 1.32 and the number of live litters (P < 0.05), and the embryonic mortality decreased significantly (P<0.05). Than, add N
The weight of the litter in the CG group increased by 18% (P<0.05), and the total placental weight and total amniotic fluid volume increased by 13% and 18%, respectively (P<0.05); however, the sow's body weight, uterine weight and corpus luteum number did not have NCG. Differences between groups and controls (Table 1).
4.2 The effect of adding 0.05% NCG on the plasma free amino acid concentration in pregnant women's 0-28 days diet for 14 days and 28 days of pregnancy
Adding NCG to the gestational sow diet, the plasma concentrations of glutamate, ornithine, arginine and protein were increased by 17%, 43%, 30%, and 14%, respectively, on the 14th day of pregnancy. <0.05); Similarly, the plasma concentrations of glutamate, ornithine, arginine, and protein were increased by 41%, 24%, 39%, and 17% (P<0.05) on the 28th day of pregnancy (Table). 2).
4.3 The effect of adding 0.05% NCG on plasma NO and hormone levels in 0-28 days of gestation sows
Compared with the control group, the plasma NO concentration in the NCG group increased by 49% and 55% (P<0.05) at 14 days and 28 days, respectively. There was no difference in plasma estradiol and progesterone concentrations between the 14th and 28th day of pregnancy (Table). 3).
　 4.4 Adding NCG to the metabolism of carbohydrates, lipids and protein-related proteins
Other experimental results showed that a total of 59 proteins in the NCG-added test group showed at least a 2-fold difference (P < 0.05), including 52 high-abundance proteins and 7 low-abundance proteins. These differentially expressed proteins are mainly involved in physiological responses such as cell adhesion, energy metabolism, lipid metabolism, protein metabolism, anti-oxidative stress and immune response. Adding NCG to the feed, several proteins closely related to embryo implantation and development at 14 days of sow pregnancy, such as integrin αv, integrin β3, cytoskeletal protein talin, and endothelial nitric oxide synthase, etc. Proteins have up-regulated expression in varying degrees.
5. Conclusion:
The results of this study provide a theoretical basis for the changes in protein abundance of endometrium induced by NCG supplementation in sow diets, and are highly correlated with embryo survival and development.
　 Note: This article is excerpted from "Maternal N-Carbamylglutamate Supplementation during Early Pregnancy Enhances Embryonic Survival and Development through Modulation of the Endometrial Proteome in Gilts", The Journal of Nutrition, 2015, 145, 2212-20
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