STUDY OF THE ANTIOXIDANT ACTIVITY OF SHEEP WHEY PROTEIN IN
VITRO AND IN C2C12 CELL LINE
Efthalia kerasioti, Alexandros Priftis, Stefanos Aivazidis, Dimitrios Stagos,
Demetrios Kouretas
Department of Biochemistry & Biotechnology, University of Thessaly, Larisa,
41221
Introduction. Whey is considered as by-product of cheese manufacturing, but it is
also described as functional food (1, 2). Whey is the liquid that remains in an
aqueous environment after removal of casein. We examined 1) in vitro the
scavenging activity of sheep whey protein and compared with that one of beef
protein, soy protein and cow whey protein and 2) its antioxidant properties in C2C12
muscle cell line.
Methodology. It was determined in vitro the ability of proteins to neutralize DPPH•,
ABTS•+, OH•, O2·- radicals as well as their reducing power. Also, we determined the
levels of glutathione (GSH) and free radicals (ROS) by flow cytometry, as well as the
levels of thiobarbituric reactive substances (TBARS) (marker of lipid peroxidation)
spectrophotometrically in muscle cell line C2C12 under the influence of the oxidative
agent t-BOOH.
Results. All proteins showed a dose-dependent free radical scavenging activity. The
IC50 value was determined, showing the concentration that has the ability to
scavenge radical by 50% while for reducing power assay, it was determined the
RP0.5AU, the concentration that produces an absorbance of 0.5 at 700 nm (Table 1).
Table1. IC50 (mg/ml) and RP0.5AU (mg/ml) for sheep whey protein, beef protein, soy protein and
cow whey protein. Values are presented as mean±sem.
DPPH•
ABTS•+
OH•
O2·-
ΑΔ
3,1±0,06
4,1±0,15
1,8±0,20
-
1,3±0,05
Beef protein
1,3±0,12
0,6±0,02
0,85±0,05
2,5±0,2
0,9±0,03
Soy protein
2,2±0,31
3,1±1,00
1,1±0,34
-
3,1±1,20
Cow
8,2±0,55
3,9±0,09
1,7±0,10
-
4,8±0,02
Proteins
Sheep
whey
protein
whey
protein
As regards the muscle cell line C2C12, sheep whey protein protected from the
oxidative agent t-BOOH. Specifically, sheep whey protein at concentrations 0.78,
1.56, 3.12 and 6.24 mg/ml, increased GSH levels by 25%, 112%, 118% και 138%
respectively, lowered TBARS levels by 21%, 15%, 25% και 24% respectively and
decreased ROS levels by 2.8%, 12.8%, 16.4% και 41.4% respectively, compared to
t-BOOH treatment alone.
References
1)Kerasioti E, Kiskini A, Veskoukis A, Jamurtas A, Tsitsimpikou C, Tsatsakis AM,
Koutedakis Y, Stagos D, Kouretas D, Vaios K. Effect of a special carbohydrateprotein cake on oxidative stress markers after exhaustive cycling in humans. Food
Chem Toxicol. 2012 50(8):2805-2810.
2) Kerasioti E, Stagos D, Jamurtas A, Kiskini A, Koutedakis Y, Goutzourelas N,
Pournaras S, Tsatsakis AM, Kouretas D. Anti-inflammatory effects of a special
carbohydrate-whey protein cake after exhaustive cycling in humans. Food Chem
Toxicol. 2013 13:61-66.
This work was co-financed by the European Union
(European Social Fund – ESF) and Greek national
funds through the Operational Program ‘‘Education and
Lifelong Learning’’ of the National Strategic Reference
Framework (NSRF) – Research Funding Program:
Heracleitus II. Investing in knowledge society through
the European Social Fund.