日粮中添加亮氨酸对生长大鼠血浆瘦素水平和骨骼肌蛋白质代谢的影响

试验旨在探讨添加亮氨酸的日粮对生长大鼠生长性能、骨骼肌蛋白质代谢和瘦素分泌的影响。试验选用24只4周龄雄性Sprague-Dawley大鼠作为试验动物,分别饲喂添加3%L-亮氨酸的试验日粮和添加2.04%L-丙氨酸的对照日粮,试验期为12 d。试验第12天,所有大鼠大剂量一次性腹腔注射L-[ring-2H5]苯丙氨酸注射液。结果表明:日粮添加亮氨酸对大鼠生长性能没有显著影响(P>0.05);试验组大鼠血浆中瘦素浓度显著高于对照组(P<0.01)。持续饲喂试验日粮的大鼠腓肠肌和比目鱼肌的蛋白质分解均显著低于对照组大鼠(P<0.01),但仅腓肠肌蛋白质合成显著高于对照组(P<0.01)。总之,长期饲喂添加L-亮氨酸的日粮可以提高生长大鼠血浆中瘦素浓度,并可显著调节生长大鼠骨骼肌蛋白质代谢。     

亮氨酸调节动物骨骼肌蛋白质分解的研究进展

随着对亮氨酸调节试验动物(如鼠)生理功能研究的深入,学者们发现,亮氨酸对于骨骼肌蛋白质沉积的影响不仅是其可以提高骨骼肌蛋白质的合成,还由于其可以减少骨骼肌蛋白质的分解,而这一过程中,亮氨酸对于骨骼肌细胞内蛋白质降解的多条途径都存在抑制作用。本文综述了亮氨酸对动物(如鼠)骨骼肌及其细胞内蛋白质分解的调节作用,并分析讨论了这一过程当中亮氨酸的相关作用机制。     

Investigation of the Effects of a Dietary Supplement on Insulin and Adipokine Concentrations in Equine Metabolic Syndrome/Insulin Dysregulation

High insulin concentrations are a common clinical feature of equine metabolic syndrome (EMS) and insulin dysregulation. Hyperinsulinemia can induce laminitis, so reduction of insulin concentrations in response to an oral challenge should decrease risk. In human studies, diets containing a polyphenol (resveratrol) led to improvements in insulin sensitivity. In rodents, the addition of leucine to a resveratrol supplement caused a decrease in the amount of resveratrol needed to achieve a clinical effect. We hypothesize a supplementation with a low dose of a synergistic polyphenol and amino acid blend including leucine (SPB+L) would improve metabolic health in EMS/insulin dysregulated horses. Fifteen EMS/ID horses received a high or low dose of SPB+ L daily for 6 weeks. Insulin during an oral sugar test (OST), body condition score, weight, baseline high-molecular-weight (HMW) adiponectin, triglycerides, nonesterified fatty acids, and tumor necrosis factor alpha were assessed before supplementation (PRE) and after supplementation (POST) via paired Student’s t-tests and a repeated-measures mixed-model analysis of variance (significant at P < .05). There were no differences between doses. Horses in the POST group weighed significantly less, had significantly higher baseline HMW adiponectin concentrations, and had significantly lower insulin concentrations at 60- and 75-minute time points (P < .05). Insulin concentrations of the horsesin the POST group, but not in the PRE group, were lower and similar to results from the study conducted three years before the present study (PRIOR) for 0- and 60-minute time points (P < .002). An increased HMW adiponectin level supports increasing insulin sensitivity after supplementation. These results suggest that SPB + L supplementation at either dose leads to improvements in the clinical manifestations of EMS/insulin dysregulation, potentially reducing laminitis risk.     

新型萘酰亚胺衍生物合成研究

合成了一系列新型的萘酰亚胺-亮氨酸加合物,利用红外光谱、核磁共振氢／碳谱和高分辨质谱等对其结构进行了确认,并利用MTT法和SRB法对其抗肿瘤活性进行了研究。结果表明,该系列化合物对所测试的6种肿瘤细胞株（HeLa、A549、P388、HL-60、MCF-7、A375）表现出较好的增长抑制活性,IC50值为10^-6～10^-5M。     

Amino acid absorption and protein synthesis responses of turbot Scophthalmus maximus to lysine and leucine in free,dipeptide and tripeptide forms

The study aimed to investigate the effects of crystalline Lys and Leu, Lys‐Leu dipeptide and Lys‐Leu‐Lys/Leu‐Lys‐Leu tripeptide, on growth, postprandial Lys and Leu concentrations, gene expression of peptide and amino acid (AA) transporters and related gene expression of protein synthesis pathway in turbot. Three diets (CAA, Di and Tri) contained Lys and Leu as free (Lys and Leu), dipeptide (Lys‐Leu) and tripeptide (Lys‐Leu‐Lys/Leu‐Lys‐Leu) forms, respectively. Improved growth was observed in the Di group compared with the CAA group. For peptide and AAs transporters, PepT1, B⁰AT1 and y⁺LAT2 mRNA levels were affected in proximal intestine by dietary treatments. Free Lys and Leu concentrations in the CAA group were significantly higher than that of the Di and Tri groups at 6 hr postfeeding in serum and at 2, 6 and 24 hr postfeeding in muscle. For protein synthesis pathway in muscle, Akt2, TOR and S6k1 gene expression were the highest in the Di group and the lowest in the CAA group, whereas MuRF1 relative expression was the highest in the Tri group. In conclusion, dietary Lys‐Leu dipeptide was utilized more efficiently than free Lys and Leu or Lys‐Leu‐Lys/Leu‐Lys‐Leu tripeptide for turbot by regulating AAs transport, postprandial AAs concentration and the synthesis of muscle proteins.     

Leucine‐induced promotion of post‐absorptive EAA utilization and hepatic gluconeogenesis contributes to protein synthesis in skeletal muscle of dairy calves

The high rate of protein synthesis in skeletal muscle of dairy calves can benefit their first lactation even lifetime milk yield. Since the rate of protein synthesis is relatively low in the post‐absorptive state, the aim of this research was to determine whether leucine supplementation could increase the post‐absorptive essential amino acid (EAA) utilization and protein synthesis in the skeletal muscle. Ten male neonatal dairy calves (38 ± 3 kg) were randomly assigned to either the control (CON, no leucine supplementation, n = 5) or supplementation with 1.435 g leucine/L milk (LEU, n = 5). Results showed that leucine significantly increased the length and protein concentration in longissimus dorsi (LD) muscle, whereas it decreased creatinine concentration and glutamic‐oxalacetic transaminase (GOT) activity. Compared to the control group, leucine supplementation also reduced the glutamic‐pyruvic transaminase (GPT) activity. Supplementation of leucine improved the phosphorylation of mammalian target of rapamycin (mTOR), eukaryotic initiation factor 4E‐binding protein 1 (4EBP1) and substrates ribosomal protein S6 kinase 1 (p70S6K). Supplementation of leucine resulted in increased concentrations of glucose, methionine, threonine, histidine and EAAs and decreased concentration of arginine in serum. Liver glucose concentration was higher and pyranic acid was lower in LEU compared to CON. In conclusion, leucine supplementation can promote post‐absorptive EAA utilization and hepatic gluconeogenesis, which contributes to protein synthesis in skeletal muscle of dairy calves.     

Regulation of the expression of key signalling molecules in mTOR pathway of skeletal muscle satellite cells in neonatal chicks: Effects of leucine and glycine–leucine peptide

This study was conducted to analyse the effects of leucine (Leu) and glycine (Gly)‐Leu peptide on expressions of key signalling molecules in mTOR pathway of skeletal muscle satellite cells in neonatal chicks. The 4‐day‐old male AA broilers with similar weight were selected to obtain the broiler skeletal muscle satellite cells with the two‐step method of collagenase‐I and trypsin digestion. The satellite cells were subjected to primary culture in vitro, and they were cultured in DMEM medium with the Leu concentration of 0.2 mM and 2 mM as well as with the Gly‐Leu peptide concentration of 0.2 mM and 2 mM. The experiment lasted for 5 days. The results showed that TOR, S6K1 and 4E‐BP1 mRNA expressions in the medium with Leu concentration of 2 mM were significantly higher than that in 0.2 mM group (p < 0.05). There was no difference between the medium with Gly‐Leu concentration of 2 mM and 0.2 mM on the TOR, S6K1 and 4E‐BP1 mRNA expressions (p > 0.05). In conclusion, Leu significantly increases TOR, S6K1 and 4E‐BP1 mRNA expressions of skeletal muscle satellite cells, but Gly‐Leu peptide has no effect on them.     

Improving performance of finishing pigs with added valine,isoleucine, and tryptophan: validating a meta-analysis model

Based on results of a recent meta-analysis, we hypothesized that increased dietary Val, Ile, or Trp could correct possible amino acid interactions because of excess Leu in diets containing high levels of corn protein, namely dried distiller’s grains with solubles (DDGS). A total of 1,200 pigs (PIC TR4 × (Fast LW × PIC L02); initially 33.6 ± 0.6 kg) were used in a 103-d study. The 6 dietary treatments were corn–soybean meal (SBM)-DDGS-based as follows: (1) high SBM and low level of l-Lys HCl (HSBM), (2) high l-Lys HCl and moderate Ile, Val, Trp (AA above NRC 2012 estimates; NC), (3) moderate l-Lys HCl and high Ile, Val, and Trp (PC), and PC with either increased (4) L-Val (PC+Val), (5) L-Ile (PC+Ile), or (6) L-Trp (PC+Trp). Pigs fed the NC diet were predicted to have the poorest average daily gain (ADG), the PC diet to be intermediate, and pigs fed the HSBM, PC+Val, PC+Ile, and PC+Trp have the same and highest predicted ADG. In the grower period (34 to 90 kg), ADG was greater (Ρ < 0.05) for the pigs fed HSBM and PC+Val diets than the NC with pigs fed other diets intermediate. Pigs fed HSBM were more (Ρ < 0.05) efficient (G:F) than the NC and PC with pigs fed other diets intermediate. In the finisher period (90 to 136 kg), ADG was greater (Ρ < 0.05) for pigs fed PC+Ile than that of the NC with pigs fed other diets intermediate. Pigs fed PC+Val had greater (Ρ < 0.05) average daily feed intake (ADFI) than the NC with pigs fed other diets intermediate. However, PC+Ile pigs were more (Ρ < 0.05) efficient than PC+Val with pigs fed other diets intermediate. Overall, ADG was greater (Ρ < 0.05) for pigs fed HSBM, PC+Val, and PC+Ile diets than the NC with pigs fed other diets intermediate. Pigs fed the PC+Val diet had greater (Ρ < 0.05) ADFI than the NC with pigs fed other diets intermediate. No differences were detected between treatments for overall G:F or other carcass characteristics. In conclusion, increasing Val or Ile in high l-Lys-HCl-DDGS-based diets improved growth performance compared with pigs fed diets containing high levels of l-Lys HCl without added Val and Ile. These results present evidence that the recently developed meta-analysis can predict the relative differences in overall ADG for pigs fed the NC, PC, PC+Val, and PC+Ile diets; however, the predicted G:F was less accurate. The data demonstrate that the negative effects of high Leu concentrations in corn-DDGS-based diets can be reversed by increasing the ratios of Val and Ile relative to Lys.     

A new method for determining the metabolic activity of specific bacterial populations in soil using tritiated leucine and immunomagnetic separation

 A new assay, using immunomagnetic separation and uptake of tritiated leucine ([³H]-Leu), was developed for measuring the in situ metabolic activity of specific bacterial populations in soil. Such assays are needed to assess the role individual species play in diverse microbial soil communities. The method was optimized using Pseudomonas putida KT2440 : :Tc⁺/TOL::gfp inoculated into soil microcosms. Inoculated soil samples were incubated with [³H]-Leu followed by an immunomagnetic separation to recover the target bacteria. Radiolabel incorporated by the target bacteria was then measured. Incubation time with immunomagnetic beads was not critical for optimal target cell recovery, but samples needed to be washed at least 5 times during the immunomagnetic separation to reduce unspecific binding of the indigenous soil bacteria to the magnetic beads. Soil absorption of the polyclonal antibody further reduced this unspecific binding, resulting in <1% contamination by indigenous soil bacteria relative to numbers of recovered target cells. The assay was tested by investigating the effect of different incubation temperatures on the metabolic activity of the target cells. As expected, a linear relationship between activity and temperature was observed, demonstrating the sensitivity of the assay. The method was applied to compare activities of the target strain in bulk soil and in the rhizosphere of barley. Contrary to what was anticipated, no significant difference in metabolic activity was observed. Received: 4 November 1999     

Growth rate of bacteria is affected by soil texture and extraction procedure

Effects of soil texture on the extraction efficiency of bacteria from soils and on biosynthetic activity of the extracted bacteria were studied. Bacterial extracts were prepared from three soils of different texture by homogenization (ultrasonication and mixing) or by homogenization-centrifugation at different speeds. Bacterial biosynthetic activity was estimated using thymidine and leucine incorporation techniques. In each step of the extraction procedure, a higher extractability of bacteria was obtained in finer soils than in coarse soil. Also cell-specific growth rates of bacteria were higher in the finer soils than in the coarse soil. However, in all soils, the extracted bacteria always had significantly lower cell-specific thymidine and leucine incorporation rates than the bacteria in soil slurries and thus did not represent so well the bacterial growth in the original soils. The total declines in cell-specific incorporation rates caused by the extraction were larger in fine soil (96-98%) than in coarse soil (90%), but bacteria in the coarse soil were more responsive to only minor intervention. The homogenization-centrifugation method eliminated the differences in bacterial biosynthesis found when working with soil slurries. Therefore, we recommend using of soil slurries or, optionally, soil suspensions to compare bacterial biosynthetic activity among soils of different textures.