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Glutamine supplementation stimulates cell proliferation in skeletal muscle and cultivated myogenic cells of low birth weight piglets.

ABSTRACT: Muscle growth of low birth weight (LBW) piglets may be improved with adapted nutrition. This study elucidated effects of glutamine (Gln) supplementation on the cellular muscle development of LBW and normal birth weight (NBW) piglets. Male piglets (n = 144) were either supplemented with 1 g Gln/kg body weight or an isonitrogeneous amount of alanine (Ala) between postnatal day 1 and 12 (dpn). Twelve piglets per group were slaughtered at 5, 12 and 26 dpn, one hour after injection with Bromodeoxyuridine (BrdU, 12 mg/kg). Muscle samples were collected and myogenic cells were isolated and cultivated. Expression of muscle growth related genes was quantified with qPCR. Proliferating, BrdU-positive cells in muscle sections were detected with immunohistochemistry indicating different cell types and decreasing proliferation with age. More proliferation was observed in muscle tissue of LBW-GLN than LBW-ALA piglets at 5 dpn, but there was no clear effect of supplementation on related gene expression. Cell culture experiments indicated that Gln could promote cell proliferation in a dose dependent manner, but expression of myogenesis regulatory genes was not altered. Overall, Gln supplementation stimulated cell proliferation in muscle tissue and in vitro in myogenic cell culture, whereas muscle growth regulatory genes were barely altered.

SUBMITTER: Zhao Y

PROVIDER: S-EPMC8239033 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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Glutamine supplementation stimulates cell proliferation in skeletal muscle and cultivated myogenic cells of low birth weight piglets.

Zhao Yaolu Y Albrecht Elke E Stange Katja K Li Zeyang Z Schregel Johannes J Sciascia Quentin L QL Metges Cornelia C CC Maak Steffen S

Scientific reports 20210628 1

Muscle growth of low birth weight (LBW) piglets may be improved with adapted nutrition. This study elucidated effects of glutamine (Gln) supplementation on the cellular muscle development of LBW and normal birth weight (NBW) piglets. Male piglets (n = 144) were either supplemented with 1 g Gln/kg body weight or an isonitrogeneous amount of alanine (Ala) between postnatal day 1 and 12 (dpn). Twelve piglets per group were slaughtered at 5, 12 and 26 dpn, one hour after injection with Bromodeoxyuri ...[more]

PMID: 34183762

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Project description:Our previous studies revealed that L-arginine supplementation had beneficial effects on intestinal barrier functions of low-birth-weight (LBW) piglets, which were associated with the enhanced antioxidant capacity. Moreover, mitochondrial functions are closely related to the redox state. This study was to explore potential mechanisms of L-arginine-induced beneficial effects against intestinal dysfunction by regulating mitochondrial function of LBW piglets. Twenty 4-day-old normal birth weight (NBW) piglets (BW: 2.08 ± 0.09 kg) and 20 LBW siblings (BW: 1.16 ± 0.07 kg) were artificially fed either a basal diet or a basal diet supplemented with 1.0% L-arginine for 21 d, respectively. Growth performance, intestinal morphology, redox status, mitochondrial morphology, and mitochondrial functions were examined. Data were subjected to two-way analysis of variance. LBW piglets presented lower (p < 0.05) ADG, shorter (p < 0.05) intestinal villus height, lower (p < 0.05) jejunal adenosine triphosphate (ATP) content and higher (p < 0.05) concentrations of Ca2+ and 8-OH-dG in jejunal mitochondria, compared with NBW piglets. Supplementation with 1.0% L-arginine significantly increased (p < 0.05) ADG, the activities of CAT, SOD, and GPx, intestinal villus height and mRNA abundances of ZO-1 (2-fold) in the jejunum of LBW piglets, but not in NBW piglets. Furthermore, the concentrations of ATP and the transcription of COX IV, COX V genes were up-regulated (p < 0.05) and the concentration of Ca2+ and 8-OH-dG were decreased (p < 0.05) in arginine-treated LBW piglets. The results suggest that mitochondrial morphology is affected, and mitochondrial functions are impaired in the jejunum of LBW piglets. While supplementation with 1.0% L-arginine relieved intestinal dysfunction through enhancing antioxidant capacity and improving mitochondrial functions via repairing mitochondrial morphology, normalizing mitochondrial calcium, and increasing ATP concentration in the jejunum of LBW piglets. However, supplementation with L-arginine has no significant beneficial effects on intestinal health in NBW piglets.

Project description:Piglets with low birth weight (LBW) usually have reduced muscle mass and increased lipid deposition compared with their normal-birth-weight (NBW) littermates. Supplementation of piglets with amino acids during the first days of life may improve muscle growth and simultaneously alter the intramuscular lipid deposition. The aim of the current study was to investigate the influence of glutamine (Gln) supplementation during the early suckling period on lipid deposition in the longissimus muscle (MLD) and the role of different perilipin (PLIN) family members in this process. Four groups were generated consisting of 72 male LBW piglets and 72 NBW littermates. Piglets were supplemented with either 1 g Gln/kg body weight or an isonitrogenous amount of alanine (Ala) between days post natum (dpn) 1 and 12. Twelve piglets per group were slaughtered at 5, 12, and 26 dpn, and muscle tissue was collected. Perilipins were localized by immunohistochemistry in muscle sections. The mRNA and protein abundances of PLIN family members and related lipases were quantified by quantitative RT-PCR (qPCR) and western blots, respectively. While PLIN1 was localized around lipid droplets in mature and developing adipocytes, PLIN2 was localized at intramyocellular lipid droplets, PLIN3 and 4 at cell membranes of muscle fibers and adipocytes, and PLIN5 in the cytoplasm of undefined cells. The western blot results indicated higher protein abundances of PLIN2, 3, 4, and 5 in LBW piglets (p < 0.05) at 5 dpn compared with their NBW littermates independent of supplementation, while not directly reflecting the mRNA expression levels. The mRNA abundance of PLIN2 was lower while PLIN4 was higher in piglets at 26 dpn in comparison with piglets at 5 dpn (p < 0.01). Relative mRNA expression of LPL and CGI-58 was lowest in piglets at 5 dpn (p < 0.001). However, ATGL mRNA was not influenced by birth weight or supplementation, but the Spearman correlation coefficient analysis revealed close correlations with PLIN2, 4, and 5 mRNA at 5 and 26 dpn (r > 0.5, p < 0.001). The results indicated the importance of birth weight and age for intramuscular lipid deposition and different roles of PLIN family members in this process, but no clear modulating effect of Gln supplementation.

Project description:Background:Low-birth-weight (LBW) neonates are susceptible to intestinal dysfunction. Furthermore, the antioxidant capacity of LBW neonates is significantly lower compared with that of normal-birth-weight (NBW) neonates both at birth and at weaning. In LBW neonates, dietary supplementation with arginine has shown beneficial effects on intestinal function. Objective:The present study explored the potential mechanisms of arginine-induced protective effects against intestinal dysfunction in LBW piglets. Methods:Forty 4-d-old LBW piglets [body weight (BW): 1.05 ± 0.04 kg] (Large White × Landrace) were assigned to 4 treatments and artificially fed a whole-milk powder- and whey protein concentrate-based diet (containing 0.65% arginine) either not supplemented with arginine (LBWC) or supplemented with 0.5%, 1.0%, or 1.5% l-arginine for 21 d. In addition, 10 NBW siblings (BW: 1.96 ± 0.03 kg) were selected and fed the basal diet. Growth performance, intestinal morphology, mRNA expression of tight junction protein, redox-sensitive genes and nitric oxide (NO) synthase, cytokines, and redox indexes were determined. Data were subjected to 1-factor ANOVA. Results:LBW piglets exhibited poorer growth performance (29.9%), lower Claudin1 mRNA level (63.6%), lower antioxidant capacity (22.9 ? 24.3%), and higher jejunum interleukin 1 (IL-1) concentration (18.8%) compared with NBW piglets. Dietary supplementation with 0.5% and 1.0% l-arginine significantly enhanced daily BW gain of LBW piglets by 13.6% and 18.2%, respectively. Compared with LBWC, dietary supplementation with 1.0% l-arginine increased the serum insulin concentration (32.2%) and villus height in the jejunum (12.2%) and ileum (20.5%). In the jejunum, the mRNA levels for Claudin1 (105%) and glutathione peroxidase (36%) were higher, and the concentrations of IL-1 (31.7%) and tumor necrosis factor ? (TNF-?) (30%) were lower in arginine-treated piglets than in the LBWC group. However, NO synthase activity and NO concentration in the jejunum of LBW piglets were not influenced by l-arginine supplementation. Conclusion:The results suggested that supplementation with 1.0% l-arginine not only promoted growth performance and improved intestinal functions in LBW piglets but also improved intestinal barrier functions and enhanced antioxidant capacity by an NO-independent pathway.

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Glutamine supplementation stimulates cell proliferation in skeletal muscle and cultivated myogenic cells of low birth weight piglets.

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Glutamine supplementation stimulates cell proliferation in skeletal muscle and cultivated myogenic cells of low birth weight piglets.

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