Grass-Fed and Non-Grass-Fed Whey Protein Supplementation and High-Intensity Eccentric Exercise Do Not Affect Arterial Stiffness and Systemic Hemodynamics in Resistance-Trained Individuals Original Research
Main Article Content
Keywords
EIMD, squat, cardiovascular
Abstract
Introduction: Dairy products from pasture-raised grass-fed cows are known for their higher anti-inflammatory and antioxidant content, which may enhance vascular modulation compared to products from grain-fed cows. Given that eccentric muscle loading (EML) and the resulting exercise-induced muscle damage (EIMD) may temporarily disrupt vascular homeostasis, this study tested the hypothesis that whey protein from pasture-raised grass-fed cows (PRWP) would improve markers of vascular recovery compared to conventional whey protein (CWP) or a non-protein control (NPC).
Methods: A randomized, double-blind, placebo-controlled study was used to test 39 resistance-trained participants (66% male, PRWP, n = 14; CWP, n = 12; NPC, n = 13). EIMD was induced via an eccentric barbell back squat protocol, followed by vascular assessments (peripheral/central blood pressure and carotid-femoral pulse wave velocity using applanation tonometry) at 24, 48, and 72 hours post-EML.
Results: No between-group differences in blood pressure or vascular measures were observed pre-EML. However, a significant interaction was observed 48 hours post-EML, with diastolic blood pressure (DBP) elevated in the PRWP group (PRWP, 77 ± 11 mm Hg; CWP, 66 ± 9 mm Hg; NPC, 67 ± 10 mm Hg; P = 0.011). Pulse wave velocity did not differ significantly between groups (P = 0.598), visit (P = 0.753), or group-by-visit interaction (P = 0.418). No additional differences in blood pressure or vascular assessments were observed post-EML.
Conclusions: Unexpectedly, PRWP increased DBP 48 hours post-EML, without any observed benefit in vascular recovery markers. Participants' training backgrounds may have reduced the sensitivity to detect EML-induced vascular disruptions.
References
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2. Barnes JN, Trombold JR, Dhindsa M, Lin HF, Tanaka H. Arterial stiffening following eccentric exercise-induced muscle damage. J Appl Physiol. 2010;109(4):1102–8.
3. Burr JF, Boulter M, Beck K. Arterial stiffness results from eccentrically biased downhill running exercise. J Sci Med Sport. 2015 Mar 1;18(2):230–5.
4. Caldwell JT, Wardlow GC, Branch PA, Ramos M, Black CD, Ade CJ. Effect of exercise‐induced muscle damage on vascular function and skeletal muscle microvascular deoxygenation. Physiol Rep. 2016;4(22):e13032.
5. Choi Y, Akazawa N, Zempo-Miyaki A, Ra SG, Shiraki H, Ajisaka R, et al. Acute effect of high-intensity eccentric exercise on vascular endothelial function in young men. J Strength Cond Res. 2016 Aug;30(8):2279–85.
6. Larsen RG, Thomsen JM, Hirata RP, Steffensen R, Poulsen ER, Frøkjær JB, et al. Impaired microvascular reactivity after eccentric muscle contractions is not restored by acute ingestion of antioxidants or dietary nitrate. Physiol Rep. 2019;7(13):e14162.
7. Lin HF, Chou CC, Cheng HM, Tanaka H. Delayed onset vascular stiffening induced by eccentric resistance exercise and downhill running. Clin J Sport Med. 2017;27(4):369–74.
8. Hunter GR, Fisher G, Bryan DR, Borges JH, Carter SJ. Divergent blood pressure response following high-intensity interval exercise: a signal of delayed recovery? J Strength Cond Res. 2018;32(11):3004.
9. Carter SJ, Goldsby TU, Fisher G, Plaisance EP, Gower BA, Glasser SP, et al. Systolic blood pressure response after high-intensity interval exercise is independently related to decreased small arterial elasticity in normotensive African American women. Appl Physiol Nutr Metab Physiol Appl Nutr Metab. 2016 May;41(5):484–90.
10. Davies RW, Carson BP, Jakeman PM. The effect of whey protein supplementation on the temporal recovery of muscle function following resistance training: A systematic review and meta-analysis. Nutrients. 2018;10(2):221.
11. Pearson AG, Hind K, Macnaughton LS. The impact of dietary protein supplementation on recovery from resistance exercise-induced muscle damage: A systematic review with meta-analysis. Eur J Clin Nutr. 2023 Aug;77(8):767–83.
12. Price D, Jackson KG, Lovegrove JA, Givens DI. The effects of whey proteins, their peptides and amino acids on vascular function. Nutr Bull. 2022;47(1):9–26.
13. Fekete ÁA, Givens DI, Lovegrove JA. The impact of milk proteins and peptides on blood pressure and vascular function: a review of evidence from human intervention studies. Nutr Res Rev. 2013 Dec;26(2):177–90.
14. Vajdi M, Musazadeh V, Zareei M, Adeli S, Karimi A, Hojjati A, et al. The effects of Whey protein on blood pressure: A systematic review and dose-response meta-analysis of randomized controlled trials. Nutr Metab Cardiovasc Dis. 2023;
15. Taddei S, Bortolotto L. Unraveling the Pivotal Role of Bradykinin in ACE Inhibitor Activity. Am J Cardiovasc Drugs Drugs Devices Interv. 2016 Oct;16(5):309–21.
16. Oliveira GV de, Volino-Souza M, Cordeiro EM, Conte-Junior CA, Alvares TS. Effects of fish protein hydrolysate ingestion on endothelial function compared to whey protein hydrolysate in humans. Int J Food Sci Nutr. 2020 Feb 17;71(2):242–8.
17. Chamata Y, Watson KA, Jauregi P. Whey-derived peptides interactions with ACE by molecular docking as a potential predictive tool of natural ACE inhibitors. Int J Mol Sci. 2020;21(3):864.
18. Hussein FA, Chay SY, Zarei M, Auwal SM, Hamid AA, Ibadullah W, et al. Effects of hydrolyzed whey versus other whey protein supplements on the physiological response to 8 weeks of resistance exercise in college-aged males. Foods. 2020;9(1):64.
19. Barenie MJ, Escalera A, Carter SJ, Grange HE, Paris HL, Krinsky D, et al. Grass-Fed and Non-Grass-Fed Whey Protein Consumption Do Not Attenuate Exercise-Induced Muscle Damage and Soreness in Resistance-Trained Individuals: A Randomized, Placebo-Controlled Trial. J Diet Suppl. 2023 Nov 20;1–30.
20. Elsworthy N, Callaghan DE, Scanlan AT, Kertesz AHM, Kean CO, Dascombe BJ, et al. Validity and reliability of using load-velocity relationship profiles to establish back squat 1 m·s‐1 load. J Strength Cond Res. 2021 Feb;35(2):340–6.
21. Romero-Parra N, Barba-Moreno L, Rael B, Alfaro-Magallanes VM, Cupeiro R, Díaz ÁE, et al. Influence of the menstrual cycle on blood markers of muscle damage and inflammation following eccentric exercise. Int J Environ Res Public Health. 2020;17(5):1618.
22. Berger RA. Determination of the resistance load for 1-RM and 10-RM. J Assoc Phys Ment Rehabil. 1961;15:108–10.
23. Subar AF, Kirkpatrick SI, Mittl B, Zimmerman TP, Thompson FE, Bingley C, et al. The automated self-administered 24-hour dietary recall (ASA24): a resource for researchers, clinicians and educators from the national cancer institute. J Acad Nutr Diet. 2012 Aug;112(8):1134–7.
24. Van Bortel LM, Laurent S, Boutouyrie P, Chowienczyk P, Cruickshank JK, De Backer T, et al. Expert consensus document on the measurement of aortic stiffness in daily practice using carotid-femoral pulse wave velocity. J Hypertens. 2012 Mar;30(3):445–8.
25. Oppliger RA, Magnes SA, Popowski LA, Gisolfi CV. Accuracy of urine specific gravity and osmolality as indicators of hydration status. Int J Sport Nutr Exerc Metab. 2005;15(3):236–51.
26. Maruyama N, Asai T, Abe C, Inada A, Kawauchi T, Miyashita K, et al. Establishment of a highly sensitive sandwich ELISA for the N-terminal fragment of titin in urine. Sci Rep. 2016;6(1):1–11.
27. Banyard HG, Nosaka K, Sato K, Haff GG. Validity of various methods for determining velocity, force, and power in the back squat. Int J Sports Physiol Perform. 2017;12(9):1170–6.
28. Butlin M, Qasem A. Large Artery Stiffness Assessment Using SphygmoCor Technology. Pulse. 2016 Dec 1;4(4):180–92.
29. Sun ZDY, Cheriyan J. Non-invasive measurements of arterial function: what? when? why should we use them? Heart. 2019;105(15):1203–11.
30. Hwang MH, Yoo JK, Kim HK, Hwang CL, Mackay K, Hemstreet O, et al. Validity and reliability of aortic pulse wave velocity and augmentation index determined by the new cuff-based SphygmoCor Xcel. J Hum Hypertens. 2014 Aug;28(8):475–81.
31. Pauca AL, O’Rourke MF, Kon ND. Prospective evaluation of a method for estimating ascending aortic pressure from the radial artery pressure waveform. Hypertens Dallas Tex 1979. 2001 Oct;38(4):932–7.
32. Karamanoglu M, O’ROURKE MF, Avolio AP, Kelly RP. An analysis of the relationship between central aortic and peripheral upper limb pressure waves in man. Eur Heart J. 1993;14(2):160–7.
33. Townsend RR, Wilkinson IB, Schiffrin EL, Avolio AP, Chirinos JA, Cockcroft JR, et al. Recommendations for improving and standardizing vascular research on arterial stiffness: a scientific statement from the American Heart Association. Hypertension. 2015;66(3):698–722.
34. Pearcey GE, Bradbury-Squires DJ, Kawamoto JE, Drinkwater EJ, Behm DG, Button DC. Foam rolling for delayed-onset muscle soreness and recovery of dynamic performance measures. J Athl Train. 2015;50(1):5–13.
35. Fernandes JFT, Lamb KL, Twist C. Exercise-induced muscle damage and recovery in young and middle-aged males with different resistance training experience. Sports. 2019 Jun;7(6):132.
36. Paulsen G, Mikkelsen U, Raastad T, Peake J. Leucocytes, cytokines and satellite cells: What role do they play in muscle damage and regeneration following eccentric exercise? Exerc Immunol Rev. 2012 Aug 10;18:42–97.
37. Lin HF, Chou CC, Cheng HM, Tanaka H. Delayed Onset Vascular Stiffening Induced by Eccentric Resistance Exercise and Downhill Running. Clin J Sport Med Off J Can Acad Sport Med. 2017 Jul;27(4):369–74.
38. Beckmann JS, Spencer M. Calpain 3, the “gatekeeper” of proper sarcomere assembly, turnover and maintenance. Neuromuscul Disord NMD. 2008 Dec;18(12):913–21.
39. Yamaguchi S, Suzuki K, Kanda K, Okada J. N-terminal fragments of titin in urine as a biomarker for eccentric exercise-induced muscle damage. J Phys Fit Sports Med. 2020;9(1):21–9.
40. Yamaguchi S, Suzuki K, Inami T, Kanda K, Hanye Z, Okada J. Changes in Urinary Titin N-terminal Fragment Concentration after Concentric and Eccentric Exercise. J Sports Sci Med. 2020 Feb 24;19(1):121–9.
41. Inami T, Yamaguchi S, Ishida H, Kohtake N, Morito A, Yamada S, et al. Changes in Muscle Shear Modulus and Urinary Titin N-Terminal Fragment after Eccentric Exercise. J Sports Sci Med. 2022 Dec 1;21(4):536–44.
42. Tanabe Y, Shimizu K, Kondo E, Yasumatsu M, Nakamura D, Sagayama H, et al. Urinary N-terminal fragment of titin reflects muscle damage after a soccer match in male collegiate soccer players. J Strength Cond Res. 2021;35(2):360–5.
43. García-Mateo P, Ramirez-Campillo R, García-De-Alcaraz A, Rodríguez-Pérez M. A meta-analysis of the effects of strength training on arterial stiffness. Hum Mov [Internet]. 2022 [cited 2023 Mar 3];24(1). Available from: https://www.termedia.pl/A-meta-analysis-of-the-effects-of-strength-training-on-arterial-stiffness,129,47243,0,1.html
44. Karanasios E, Ryan-Stewart H, Faulkner J. The acute effects of resistance training on arterial stiffness: A systematic review. J Trainology. 2023;12(1):5–13.
45. Kresnajati S, Lin YY, Mündel T, Bernard JR, Lin HF, Liao YH. Changes in arterial stiffness in response to various types of exercise modalities: a narrative review on physiological and endothelial senescence perspectives. Cells. 2022 Nov 9;11(22):3544.
46. Au JS, Oikawa SY, Morton RW, Macdonald MJ, Phillips SM. Arterial stiffness is reduced regardless of resistance training load in young men. Med Sci Sports Exerc. 2017 Feb;49(2):342–8.
47. Lau WY, Blazevich AJ, Newton MJ, Wu SSX, Nosaka K. Reduced muscle lengthening during eccentric contractions as a mechanism underpinning the repeated-bout effect. Am J Physiol-Regul Integr Comp Physiol. 2015;308(10):R879–86.
48. The Reference Values for Arterial Stiffness’ Collaboration. Determinants of pulse wave velocity in healthy people and in the presence of cardiovascular risk factors: ‘establishing normal and reference values.’ Eur Heart J. 2010 Oct 1;31(19):2338–50.
49. Stacy MR, Bladon KJ, Lawrence JL, McGlinchy SA, Scheuermann BW. Serial assessment of local peripheral vascular function after eccentric exercise. Appl Physiol Nutr Metab. 2013 May 28;38(12):1181–6.
50. Yiğit A, Bielska P, Cais-Sokolińska D, Samur G. Whey proteins as a functional food: Health effects, functional properties, and applications in food. J Am Nutr Assoc. 2023 Feb 1;1–11.
51. Fekete AA, Giromini C, Chatzidiakou Y, Givens DI, Lovegrove JA. Whey protein lowers blood pressure and improves endothelial function and lipid biomarkers in adults with prehypertension and mild hypertension: results from the chronic Whey2Go randomized controlled trial. Am J Clin Nutr. 2016;104(6):1534–44.
52. Ballard KD, Kupchak BR, Volk BM, Mah E, Shkreta A, Liptak C, et al. Acute effects of ingestion of a novel whey-derived extract on vascular endothelial function in overweight, middle-aged men and women. Br J Nutr. 2013;109(5):882–93.
53. Yang J, Wang HP, Tong X, Li ZN, Xu JY, Zhou L, et al. Effect of whey protein on blood pressure in pre‐and mildly hypertensive adults: A randomized controlled study. Food Sci Nutr. 2019;7(5):1857–64.
54. Rontoyanni VG, Werner K, Sanders TA, Hall WL. Differential acute effects of carbohydrate-and protein-rich drinks compared with water on cardiac output during rest and exercise in healthy young men. Appl Physiol Nutr Metab. 2015;40(8):803–10.
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Feb 16, 2026
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Grass-Fed and Non-Grass-Fed Whey Protein Supplementation and High-Intensity Eccentric Exercise Do Not Affect Arterial Stiffness and Systemic Hemodynamics in Resistance-Trained Individuals: Original Research. (2026). Research in Strength and Performance, 6(1). https://doi.org/10.53520/rsp2026.105178
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Copyright, 2026 by the authors. This work is licensed under the Creative Commons Attribution 4.0 International License.