Glucocorticoid-induced osteoporosis (GIO) is one of the serious side effects which have become the most common secondary osteoporosis. The study aimed to evaluate the effect of different levels of Lepidium sativum seeds and buttermilk on glucocorticoid-induced osteoporosis in rats. Methods: The present study was carried out in the National Nutrition Institute NNI Cairo, on forty-eight female rats. The rats were divided into eight groups (six rats each). The first group was fed on a basal diet and represents the negative control, while the other seven groups were injected subcutaneously with betamethasone at a dose of 4 mg/ kg BW three times a week. One group of them represents the positive control. The other six groups were fed on a basal diet containing 10%, 20% Lepidium sativum seeds (LSs), 10%, 20% buttermilk (BM), 10% and 20% mixture of LSs and BM for a period of eight weeks. Results: The positive control group showed a significant decrease in serum Estradiol (E2), Calcium (Ca), Phosphorus (P) levels and Ca and P in femur bone and a significant decrease in total Bone Mineral Density (BMD), and a significant increase in serum Alkaline phosphatase (ALP).  On the other hand, all osteoporosis groups administrated with different levels of LSs and BM (10 and 20%) had a significant decrease in (ALP) and a significant increase in Ca, P in serum and Ca, P in bone and BMD, compared with the positive control group. The pathological examination of bone confirmed these results. Conclusion: Lepidium sativum seeds and buttermilk showed bone protection against glucocorticoid-induced osteoporosis in rats. Lepidium sativum seeds had a potent protective effect more than Buttermilk due to its content of isoflavones.