#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

A cell-based evaluation of a non-essential amino acid formulation as a non-bioactive control for activation and stimulation of muscle protein synthesis using ex vivo human serum


Autoři: Bijal Patel aff001;  Martina Pauk aff001;  Miryam Amigo-Benavent aff001;  Alice B. Nongonierma aff001;  Richard J. Fitzgerald aff001;  Philip M. Jakeman aff001;  Brian P. Carson aff001
Působiště autorů: Food for Health Ireland, University of Limerick, Limerick, Ireland aff001;  Human Sciences Research Unit, University of Limerick, Limerick, Ireland aff002;  Department of Physical Education & Sport Sciences, Faculty of Education and Health Sciences, University of Limerick, Limerick, Ireland aff003;  Centre for Intervention in Infection, Inflammation and Immunity (4i), University of Limerick, Limerick, Ireland aff004;  Department of Biological Sciences, University of Limerick, Limerick, Ireland aff005;  Health Research Institute, University of Limerick, Limerick, Ireland aff006
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0220757

Souhrn

Purpose

The purpose of this study was to compare the effect of treating skeletal muscle cells with media conditioned by postprandial ex vivo human serum fed with either isonitrogenous Non-Essential Amino Acid (NEAA) or a whey protein hydrolysate (WPH) on stimulating Muscle Protein Synthesis (MPS) in C2C12 skeletal muscle cells.

Methods

Blood was taken from six young healthy males following overnight fast (fasted) and 60 min postprandial (fed) ingestion of either WPH or NEAA (0.33 g.kg-1 Body Mass). C2C12 myotubes were treated with media conditioned by ex vivo human serum (20%) for 4 h. Activation of MPS signalling (phosphorylation of mTOR, P70S6K and 4E-BP1) were determined in vitro by Western Blot and subsequent MPS were determined in vitro by Western Blot and surface sensing of translation technique (SUnSET) techniques, respectively.

Results

Media conditioned by NEAA fed serum had no effect on protein signalling or MPS compared to fasted, whereas media conditioned by WPH fed serum significantly increased mTOR (Ser2448), P70S6K and 4E-BP1 phosphorylation (p<0.01, p<0.05) compared to fasted serum. Furthermore, the effect of media conditioned by WPH fed serum on protein signalling and MPS was significantly increased (p<0.01, p<0.05) compared to NEAA fed serum.

Conclusion

In summary, media conditioned by NEAA fed serum did not result in activation of MPS. Therefore, these in vitro findings suggest the use of isonitrogenous NEAA acts as an effective control for comparing bioactivity of different proteins on activation of MPS.

Klíčová slova:

Ingestion – Insulin – Muscle proteins – Phosphorylation – Serum proteins – Skeletal muscles – Muscle protein synthesis – Leucine


Zdroje

1. Borsheim E, Tipton KD, Wolf SE, Wolfe RR (2002) Essential amino acids and muscle protein recovery from resistance exercise. Am J Physiol Endocrinol Metab 283: E648–657. doi: 10.1152/ajpendo.00466.2001 12217881

2. Smith K, Reynolds N, Downie S, Patel A, Rennie MJ (1998) Effects of flooding amino acids on incorporation of labeled amino acids into human muscle protein. Am J Physiol 275: E73–78. doi: 10.1152/ajpendo.1998.275.1.E73 9688876

3. Volpi E, Kobayashi H, Sheffield-Moore M, Mittendorfer B, Wolfe RR (2003) Essential amino acids are primarily responsible for the amino acid stimulation of muscle protein anabolism in healthy elderly adults. Am J Clin Nutr 78: 250–258. doi: 10.1093/ajcn/78.2.250 12885705

4. Tipton KD, Gurkin BE, Matin S, Wolfe RR (1999) Nonessential amino acids are not necessary to stimulate net muscle protein synthesis in healthy volunteers. J Nutr Biochem 10: 89–95. doi: 10.1016/s0955-2863(98)00087-4 15539275

5. Morton RW, Murphy KT, McKellar SR, Schoenfeld BJ, Henselmans M, et al. (2018) A systematic review, meta-analysis and meta-regression of the effect of protein supplementation on resistance training-induced gains in muscle mass and strength in healthy adults. Br J Sports Med 52: 376–384. doi: 10.1136/bjsports-2017-097608 28698222

6. Phillips SM (2016) The impact of protein quality on the promotion of resistance exercise-induced changes in muscle mass. Nutr Metab (Lond) 13: 64.

7. Xu ZR, Tan ZJ, Zhang Q, Gui QF, Yang YM (2014) Clinical effectiveness of protein and amino acid supplementation on building muscle mass in elderly people: a meta-analysis. PLoS One 9: e109141. doi: 10.1371/journal.pone.0109141 25268791

8. Xu ZR, Tan ZJ, Zhang Q, Gui QF, Yang YM (2015) The effectiveness of leucine on muscle protein synthesis, lean body mass and leg lean mass accretion in older people: a systematic review and meta-analysis. Br J Nutr 113: 25–34. doi: 10.1017/S0007114514002475 25234223

9. EFSA Panel on Dietetic Products NaAENP (2018) Draft guidance on the scientific requirements for health claims related to muscle function and physical performance. EFSA Journal 16: 5434.

10. Bohe J, Low A, Wolfe RR, Rennie MJ (2003) Human muscle protein synthesis is modulated by extracellular, not intramuscular amino acid availability: a dose-response study. J Physiol 552: 315–324. doi: 10.1113/jphysiol.2003.050674 12909668

11. Smith K, Barua JM, Watt PW, Scrimgeour CM, Rennie MJ (1992) Flooding with L-[1-13C]leucine stimulates human muscle protein incorporation of continuously infused L-[1-13C]valine. Am J Physiol 262: E372–376. doi: 10.1152/ajpendo.1992.262.3.E372 1550230

12. Atherton PJ, Smith K, Etheridge T, Rankin D, Rennie MJ (2010) Distinct anabolic signalling responses to amino acids in C2C12 skeletal muscle cells. Amino Acids 38: 1533–1539. doi: 10.1007/s00726-009-0377-x 19882215

13. Atherton PJ, Kumar V, Selby AL, Rankin D, Hildebrandt W, et al. (2017) Enriching a protein drink with leucine augments muscle protein synthesis after resistance exercise in young and older men. Clin Nutr 36: 888–895. doi: 10.1016/j.clnu.2016.04.025 27208923

14. Koopman R, Wagenmakers AJ, Manders RJ, Zorenc AH, Senden JM, et al. (2005) Combined ingestion of protein and free leucine with carbohydrate increases postexercise muscle protein synthesis in vivo in male subjects. Am J Physiol Endocrinol Metab 288: E645–653. doi: 10.1152/ajpendo.00413.2004 15562251

15. Wilkinson DJ, Bukhari SSI, Phillips BE, Limb MC, Cegielski J, et al. (2017) Effects of leucine-enriched essential amino acid and whey protein bolus dosing upon skeletal muscle protein synthesis at rest and after exercise in older women. Clin Nutr.

16. Wilkinson DJ, Hossain T, Hill DS, Phillips BE, Crossland H, et al. (2013) Effects of leucine and its metabolite beta-hydroxy-beta-methylbutyrate on human skeletal muscle protein metabolism. J Physiol 591: 2911–2923. doi: 10.1113/jphysiol.2013.253203 23551944

17. Wilkinson SB, Kim PL, Armstrong D, Phillips SM (2006) Addition of glutamine to essential amino acids and carbohydrate does not enhance anabolism in young human males following exercise. Appl Physiol Nutr Metab 31: 518–529. doi: 10.1139/h06-028 17111006

18. Carson BP, Patel B, Amigo-Benavent M, Pauk M, Kumar Gujulla S, et al. (2018) Regulation of muscle protein synthesis in an in vitro cell model using ex vivo human serum. Exp Physiol.

19. Pennings B, Boirie Y, Senden JM, Gijsen AP, Kuipers H, et al. (2011) Whey protein stimulates postprandial muscle protein accretion more effectively than do casein and casein hydrolysate in older men. Am J Clin Nutr 93: 997–1005. doi: 10.3945/ajcn.110.008102 21367943

20. Tang JE, Moore DR, Kujbida GW, Tarnopolsky MA, Phillips SM (2009) Ingestion of whey hydrolysate, casein, or soy protein isolate: effects on mixed muscle protein synthesis at rest and following resistance exercise in young men. J Appl Physiol (1985) 107: 987–992.

21. Atherton PJ, Etheridge T, Watt PW, Wilkinson D, Selby A, et al. (2010) Muscle full effect after oral protein: time-dependent concordance and discordance between human muscle protein synthesis and mTORC1 signaling. Am J Clin Nutr 92: 1080–1088. doi: 10.3945/ajcn.2010.29819 20844073

22. McCormack WG, Cooke JP, O'Connor WT, Jakeman PM (2017) Dynamic measures of skeletal muscle dialysate and plasma amino acid concentration in response to exercise and nutrient ingestion in healthy adult males. Amino Acids 49: 151–159. doi: 10.1007/s00726-016-2343-8 27704291

23. Power-Grant O, McCormack WG, Ramia De Cap M, Amigo-Benavent M, Fitzgerald RJ, et al. (2016) Evaluation of the antioxidant capacity of a milk protein matrix in vitro and in vivo in women aged 50–70 years. Int J Food Sci Nutr 67: 325–334. doi: 10.3109/09637486.2016.1153607 26960816

24. Murphy SM, Kiely M, Jakeman PM, Kiely PA, Carson BP (2016) Optimization of an in vitro bioassay to monitor growth and formation of myotubes in real time. Biosci Rep 36.

25. Goodman CA, Mabrey DM, Frey JW, Miu MH, Schmidt EK, et al. (2011) Novel insights into the regulation of skeletal muscle protein synthesis as revealed by a new nonradioactive in vivo technique. FASEB J 25: 1028–1039. doi: 10.1096/fj.10-168799 21148113

26. Mitchell WK, Phillips BE, Hill I, Greenhaff P, Lund JN, et al. (2017) Human skeletal muscle is refractory to the anabolic effects of leucine during the postprandial muscle-full period in older men. Clin Sci (Lond) 131: 2643–2653.

27. Bohe J, Low JF, Wolfe RR, Rennie MJ (2001) Latency and duration of stimulation of human muscle protein synthesis during continuous infusion of amino acids. J Physiol 532: 575–579. doi: 10.1111/j.1469-7793.2001.0575f.x 11306673

28. Rennie MJ (2007) Exercise- and nutrient-controlled mechanisms involved in maintenance of the musculoskeletal mass. Biochem Soc Trans 35: 1302–1305. doi: 10.1042/BST0351302 17956336

29. Greenhaff PL, Karagounis LG, Peirce N, Simpson EJ, Hazell M, et al. (2008) Disassociation between the effects of amino acids and insulin on signaling, ubiquitin ligases, and protein turnover in human muscle. Am J Physiol Endocrinol Metab 295: E595–604. doi: 10.1152/ajpendo.90411.2008 18577697

30. Atherton PJ, Smith K (2012) Muscle protein synthesis in response to nutrition and exercise. J Physiol 590: 1049–1057. doi: 10.1113/jphysiol.2011.225003 22289911

31. Abdulla H, Smith K, Atherton PJ, Idris I (2016) Role of insulin in the regulation of human skeletal muscle protein synthesis and breakdown: a systematic review and meta-analysis. Diabetologia 59: 44–55. doi: 10.1007/s00125-015-3751-0 26404065

32. Wilkes EA, Selby AL, Atherton PJ, Patel R, Rankin D, et al. (2009) Blunting of insulin inhibition of proteolysis in legs of older subjects may contribute to age-related sarcopenia. Am J Clin Nutr 90: 1343–1350. doi: 10.3945/ajcn.2009.27543 19740975

33. Dickinson JM, Fry CS, Drummond MJ, Gundermann DM, Walker DK, et al. (2011) Mammalian target of rapamycin complex 1 activation is required for the stimulation of human skeletal muscle protein synthesis by essential amino acids. J Nutr 141: 856–862. doi: 10.3945/jn.111.139485 21430254


Článek vyšel v časopise

PLOS One


2019 Číslo 11
Nejčtenější tento týden
Nejčtenější v tomto čísle
Kurzy

Zvyšte si kvalifikaci online z pohodlí domova

KOST
Koncepce osteologické péče pro gynekology a praktické lékaře
nový kurz
Autoři: MUDr. František Šenk

Sekvenční léčba schizofrenie
Autoři: MUDr. Jana Hořínková

Hypertenze a hypercholesterolémie – synergický efekt léčby
Autoři: prof. MUDr. Hana Rosolová, DrSc.

Svět praktické medicíny 5/2023 (znalostní test z časopisu)

Imunopatologie? … a co my s tím???
Autoři: doc. MUDr. Helena Lahoda Brodská, Ph.D.

Všechny kurzy
Kurzy Podcasty Doporučená témata Časopisy
Přihlášení
Zapomenuté heslo

Zadejte e-mailovou adresu, se kterou jste vytvářel(a) účet, budou Vám na ni zaslány informace k nastavení nového hesla.

Přihlášení

Nemáte účet?  Registrujte se

#ADS_BOTTOM_SCRIPTS#