Effects Of Creatine Pyruvate On Lipid And Protein Metabolism In Broiler Chickens

Creatine pyruvate (CrPyr) is a new multifunctional nutrient, it makes a very good combination and complement with the function of pyruvate and creatine. The mainly biological functions of CrPyr are inhibite the loss of muscle protein, increase muscle mass and promote lipid metabolism, reduce body fat percentage and delay fatigue. Currently, the research for CrPyr focused on exercise physiology, its lipid and protein metabolism in broiler chickens has been reported rarely. In this study, a study was carried out to examine the effects of exogenous added CrPyr on lipid and protein metabolism in male AA broilers; Through the changes in protein profiles of liver and muscle by using comparative proteomics, the purpose of this is to reveal the relationship between the proteins and analyse the mechanism of CrPyr effects on lipid and protein metabolism in broiler chickens.1 Effect of CrPyr with different doses on lipid and protein metabolism in broiler chickensTo assess the effects of creatine pyruvate (CrPyr) on lipid and protein metabolism in broiler chickens, a total of 400 one day-old male birds (Arbor Acres) were randomly allocated to four groups, with each group replicated four times and each replicate involving 25 birds. The broilers were provided with a commercial diet supplemented with CrPyr at 0%,1%,5% or 10% of the diet, respectively, for a period of 3 wks ad libitum (from 22 to 42 d). The content of hepatic triglycerides (TG), blood triglycerides (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), non-esterified fatty acid (NEFA), uric acid (UA), creatine kinase (CK), glucagon (Glu), insulin (Ins), leptin (Lep), triiodothyronine (T3), thyroxin (T4), free triiodothyronine (FT3) and free thyroxin (FT4) were determined by using commercial kits. The genes of acetyl CoA carboxylase (ACC), fatty acid synthetase (FAS), peroxisome proliferators activated receptor a (PPAR-a), carnitine palmitoyl transferase-I (CPT-I) in liver and muscle insulin-like growth factor 1 (IGF-1), Myostatin, Myogenin in muscle for real-time PCR assays. Four broiler chickens per replicate from each treatment were chosen randomly and slaughtered to collect specimens of blood, liver, abdominal fat at 42 day-old. In the present study, body weight (BW) and average daily gain (ADG) of broilers decreased in 10% CrPyr group.5% or 10% CrPyr of diets decreased the abdominal fat rate (AFR, abdominal fat/live weight), serum or hepatic TG concentrations of the broilers. In contrast,5% and 10% CrPyr caused a marked increase in the relative pectoral muscle weights, serum NEFA, HDL-C, Glu, Ins or Lep contents and CK activity,. The expression of hepatic PPAR-αand CPT-I, muscle IGF-1 were significantly elevated in the 5% and 10% groups, and the Myostatin mRNA expression was reduced. It was found that supplementation with 5% CrPyr enhances lipolysis and protein synthesis, while not adversely affects growth performance in broiler chickens.2 Comparison of Pyr, CrPyr and Cr on lipid and protein metabolism in broiler chickensThe effects of Pyruvate (Pyr), Creatine Pyruvate (CrPyr) and Creatine (Cr) on lipid and protein metabolism were compared in broiler chickens. Four hundred one day-old male birds (Arbor Acres) were allocated to four groups, each of which included four replicates (25 birds per replicate). Treatments consisted of unsupplemented basal diet (0%), basal diet containing 2% Pyr, basal diet containing 5% CrPyr and basal diet containing3% Cr, respectively, for a period of 3 wks ad libitum (from 22 to 42 d). Three broiler chickens per replicate from each treatment were chosen randomly and slaughtered to collect specimens of blood, liver, abdominal fat at 42 day-old. The detected index were same as text one. The results showed that CrPyr and Pyr significantly decreased the hepatic TG and serum TC concentration. CrPyr markedly increased the relative leg muscle weights, the serum NEFA and HDL-C concentrations, while the expression of CPT-I and PPAR-a mRNA in the liver were both decidedly enhanced in the CrPyr group. The serum UA content was and the Myostatin mRNA level was reduced in the CrPyr and Cr groups. Muscle IGF-1 expression was enhanced in both the CrPyr and Cr groups. In addition, CrPyr did not alter body weight or the feed conversion ratio. These results indicate that, compared with Pyr and Cr alone, CrPyr has a bifunctional role in broiler chickens, in that it influences both lipid and protein metabolism.3 Effects of Pyr, CrPyr and Cr on the expressions of liver mitochondrial protein/ enzyme in broiler chickens The male birds (Arbor Acres) were selected to study the effect of Pyr, CrPyr and Cr on the expressions of liver mitochondrial lipid and protein metabolism-related protein/enzyme in broiler chickens. Experimental design was same to the second chapte. Mitochondrial purification process using the two-step differential centrifugation and density gradient method. The activities of galactosidase (lysome), alkaline phosphodiesterase (plasma membrane), cytochrome c oxidase (mitochondria), and lactate dehydrogenase were determined to assess the purity of mitochondrion; the activities of fumaric reductase was determined to estimate the enrichment of mitochondrion; Otherwise the morphous of mitochondrion was observed by using the MitoTracker RED. The results showed that mitochondrion was separated and purificated efficiently by using this methord in present study. Proteins were isolated by differential centrifugation and were separated by two-dimensional electrophoresis, the differentially displayed spots were obtained by pattern analysis, then the spots were further identified by MALDI-TOF-MS. Compared with the control group, CrPyr treatment group received 13 different proteins, Pyr treatment group received 17 different proteins, Cr treatment group received 11 different proteins. The results showed that, CrPyr reduced the accumulation of fatty acids by down-regulating the expression of ADRP, inhibited the expression of ATP synthase to reduce the formation of fat cells, while reduced the expression of CETP to increase HDL levels and decrease TG levels, resulting in the less fat and cholesterol deposition; CrPyr and Cr increased the expression of eIF2B, which played a key role in protein synthesis, thus promoted protein synthesis, and the effect of CrPyr was more significantly than Cr; Pyr and Cr raised the expression of ACS to promote theβ-oxidation of fatty acids and reduced fat deposition; Pyr also raised the levels of methionine aminopeptidase, ornithine aminotransferase and 60kDa heat shock protein, which effected on protein processing and transport after the termination of protein synthesis. Compared with the Pyr and Cr, CrPyr plays a more important role in lipolysis and protein synthesis, and the effect was stronger than Pyr and Cr.4 Effects of Pyr, CrPyr and Cr on the expressions of liver cytosolic protein/enzyme in broiler chickensThe male birds (Arbor Acres) were selected to study the effect of Pyr, CrPyr and Cr on the expressions of liver cytosolic lipid and protein metabolism-related protein/enzyme in broiler chickens. Experimental design was same to the second chapter. Proteins were isolated by differential centrifugation and were separated by two-dimensional electrophoresis, the differentially displayed spots were obtained by pattern analysis, then the spots were further identified by MALDI-TOF-MS. Compared with the control group, CrPyr treatment group received 16 different proteins, Pyr treatment group received 19 different proteins, Cr treatment group received 12 different proteins. The results showed that, CrPyr and Pyr up-regulated the expression of FABP and CETP in liver cytoplasm to accelerate the decomposition of fatty acids and TG, thus reducing fat deposition, the effects of CrPyr was more significantly than Pyr; CrPyr and Cr raised the expression of ApoA-IV, it could accelerate the decomposition levels of TG, then reduced fat accumulation, the effects of CrPyr was significantly than Cr; Also, CyPry increased the levels of CRT and eIF3a to promote protein synthesis, and Pyr and Cr did not have effects on protein metabolism in chicken liver. Therefore, compared with Pyr and Cr, CrPyr plays a more active and effective role in lipid and protein metabolism in broiler chickens.5 Effects of Pyr, CrPyr and Cr on the expressions of muscle protein/enzyme in broiler chickensThe male birds (Arbor Acres) were selected to study the effect of Pyr, CrPyr and Cr on the expressions of muscle metabolism-related protein/enzyme in broiler chickens. Experimental design was same to the second chapter. Proteins were isolated by homogenates and were separated by two-dimensional electrophoresis, the differentially displayed spots were obtained by pattern analysis, and then the spots were further identified by MALDI-TOF-MS. Compared with the control group, CrPyr treatment group received 11 different proteins, Pyr treatment group received 11 different proteins, Cr treatment group received 10 different proteins. The results showed that, CrPyr increased the activity of MCK, elevated the levels of creatine phosphate; it also reduced the activity of phosphorylase b kinase to decrease the decomposition of glycogen and increase the body’s energy reserves, thereby affecting the body’s energy metabolism. CrPyr up-regulated the expression of eIF2, this suggested that it could promote muscle protein synthesis, the final performance were the increased expression of muscle myosin light chain and actin. Pyr raised the expression of PK and enolase, which were the key enzyme in glycolysis, while reduced the expression of myoglobin, which played a role in the transport and storage of oxygen in muscle cells, suggesting that Pyr could help the energy recovery of the body through glycolysis under hypoxia accelerated; Pyr did not have effects on muscle protein metabolism. Cr changed the expression of myosin and actin, while increased the activity of creatine kinase, but the expression was lower than CrPyr treatment group. Comprehensive comparison shows that, CrPyr plays an active role in muscle energy metabolism and protein metabolism and the effect was significantly higher than the individual active ingredients Cr and Pyr.