Osteoporosis is characterized by bone loss density and deterioration of bone tissues, with osteoclasts typically being the only cells capable of resorbing bone. In this experiment, we aimed to investigate the effects of physical exercise, specifically treadmill use, on osteoclast activity. We used 24 male rats, randomly divided into three groups: the control group(C), the osteoporotic rats given daily 200mg/kg BW for 8 weeks in the D-gal group, and the rats in the third group given daily 200mg/kg BW with a Treadmill 25m/h daily for 5 days weekly in 8 weeks (D-gal + treadmill) group. Serum blood was drawn for biochemical analysis of PTH, Vit D, calcitonin, and minerals (Na, Ca, K, P). The results showed a significant increase of serum PTH, Vit D, calcitonin & Ca in the osteoporotic rats compared with other groups, while serum Na and K showed a significant decline in the osteoporotic rats group as compared with other groups. The impact of the treadmill was observed in a significant decrease in the level of serum PTH, Vit D, calcitonin & Ca compared with the control group. This suggests that the treadmill may have a regulatory effect on these calcium homeostasis regulators. The study concludes that D-gal affects the bone of the experimental anime by causing an imbalance in the hormones that are related to the bone and causing an imbalance in the serum mineral, which plays a critical role in Osteoporosis.