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Aim: The research aimed to investigate the effect of endurance running and T. Brumale extract on α-Syn and HSF1 in the brain and serum of healthy and diabetic rats. Methods: A total of 40 Wistar rats were randomly divided into eight groups: Control (C), Exercise (E), Control-Tuber (T), Exercise-Tuber (ET), Control-Diabetes (D), Exercise-Diabetes (ED), Control-Diabetes-Tuber (CDT), and Exercise-Diabetes-Tuber (EDT). The endurance running was carried out five times per week for five weeks. The hippocampus and the serum α-Syn and HSF1 were measured using an enzyme-linked immunosorbent assay method. Results: The brain α-Syn levels were higher in diabetic groups than in the healthy groups, but insignificantly (P ≤ 0.05). The brain α-Syn level significantly increased in the EDT group compared to the T group (P ≤ 0.05). The serum level of α-Syn in the ED group was significantly higher than in the E and D groups (P ≤ 0.05). The brain HSF1 level was significantly higher in the ED group compared to the D group (P ≤ 0.05). The gene expression of hsf1 was significantly reduced in the E group compared to the other groups and the EDT group compared to ED and CDT groups (P ≤ 0.05). Furthermore, the serum HSF1 level significantly increased in the ED group compared to the D group (P ≤ 0.05). Conclusion: The results of this study suggest that progressive endurance running may improve neuroprotective conditions in diabetic patients by increasing HSF1 in the brain.