Abstract: Senescence promotes several metabolic disorders, including insulin resistance (IR). It is known that physical exercise is an important non-pharmacological agent in the prevention and treatment of IR and consequently reducing the incidence of T2DM. Rho-kinase (Rock) has been postulated as an important protein that acts directly on the phosphorylation of IRS-1, collaborating up to 50% of glucose uptake in skeletal muscle tissue. Therefore, the aim of this study was evaluate whether the increase of glucose uptake by muscle tissue from old rats after physical exercise, also occurs due to Rock metabolism. Wistar rats were distributed into 3 groups: Sedentary Young (SY): 5 months old and sedentary rats and two groups composed by aged rats (17 months old): Old Sedentary (Old) and Old Exercised (Old Exe). The Old Exe group was submitted to a single exercise session of swimming without overload, whose protocol was composed by two sessions of three hours of exercise with 45 min of rest between sessions. Four hours after exercise the animals were divided into 3 different lots of animals (each lot was composed by all groups with 7 animals per group). A lot was submitted to the insulin tolerance test, another submitted to the glucose tolerance test and a third lot was euthanized and used to collect samples from muscle tissue for further analysis of the insulin signaling pathway and Rock metabolism through immunoprecipitation and western blotting. As expected, aged rats developed insulin resistance. This finding was, in part, due to the increase of RhoE, molecule that inhibits Rock activity and decrease of RhoA, molecule that increases Rock activity, which culminated in a lower activity of Rock and consequently lower phosphorylation of IRS-1/Akt pathway and thus lower glucose uptake, which collaborated with insulin resistance. However, after exercise, old rats reversed the framework found on Rock metabolism, such as increased levels of RhoA and Rock and reduced levels of RhoE without alterations on adipose tissue volume. Physical exercise can contribute to glucose homeostasis through Rock metabolism on aged rats. Thus, these results reveal a new mechanism by which physical exercise collaborates on glucose uptake in skeletal muscle of aged animals without the use of insulin Ver menos