Method for evaluating the ability of a composition to prevent muscle damage and fatigue; food supplement and drug

Abstract

Disclosed is a method for evaluating the ability of a chemical substance or a chemical composition to prevent muscle fatigue and damage induced by physical exertion in humans by comparing the effect the chemical substance or a chemical composition has on a combination of three biological markers in cultured human primary skeletal muscle cells.

1 . A method for evaluating the ability of a chemical substance (5) or of a chemical composition (C) to prevent muscle fatigue and muscle damage induced by physical exertion in human beings, said method comprising:
 a step a) of culturing human primary skeletal muscle cells;   a step b) of differentiating the cells obtained in step a) in order to obtain myotubes;   a step c) of bringing a chemical substance (5) or a chemical composition (C) into contact with the cell medium obtained in step b);   a step d) of bringing at least one calcium ionophore agent (AI) into contact with the cell medium obtained in step c);   a step e) of measuring the level of expression of a combination of biological markers comprising:
 at least one biological marker (M 1 ) of glycemic homeostasis, chosen from the elements of the group consisting of the myokines or cytokines produced by muscles, 
 at least one biological marker (M 2 ) of muscle lesions, chosen from creatine kinase and lactate dehydrogenase, 
 at least one biological marker (M 3 ) of ATP metabolism, chosen from the elements of the group consisting of lactate, aqueous ammonia and oxypurines, in the culture medium obtained in step d); and 
   a step f) of comparing the levels of expression of each of said three biological markers (M 1 ), (M 2 ) and (M 3 ) measured in step e) with a reference expression level for each of these three biological markers.   
     
     
         2 . The method of  claim 1  wherein, in step d), the calcium ionophore agent (AI) is selected from the elements of the group consisting of beauvericin, ionomycin and calcimycin (or A23187). 
     
     
         3 . The method of  claim 2  wherein the calcium ionophore agent (AI) is calcimycin. 
     
     
         4 . The method of  claim 1  wherein the biological marker (M 1 ) is interleukin-6, the biological marker (M 2 ) is creatine kinase, and the biological marker (M 3 ) is lactate. 
     
     
         5 . The method of  claim 2  wherein the biological marker (M 1 ) is interleukin-6, the biological marker (M 2 ) is creatine kinase, and the biological marker (M 3 ) is lactate. 
     
     
         6 . The method of  claim 3  wherein the biological marker (M 1 ) is interleukin-6, the biological marker (M 2 ) is creatine kinase, and the biological marker (M 3 ) is lactate.