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Objective:To systematically evaluate the protective effects of Humulus lupulus L.extract(HLE)on osteoporosis mice.Methods:In vivo experiment,a total of 35 12-week-old female ICR mice were equally divided into 5 groups:the sham control group(sham);the ovariectomy with vehicle group(OVX);the OVX with estradiol valerate[EV,0.2 mg/(kg-d)];the OVX with low-or high-dose HLE groups[HLE,1 g/(kg·d)and 3 g/(kg·d)],7 in each group.Treatment began 1 week after the ovariectomized surgery and lasted for 12 weeks.Bone mass and trabecular bone mircoarchitecture were evaluated by micro computed tomography,and bone turnover markers in serum were evaluated using enzyme-linked immunosorbent assay(ELISA)kits.In vitro experiment,osteoblasts and osteoclasts were treated with HLE at doses of 0,4,20 and 100 μg/mL.Biomarkers for bone formation in osteoblasts and bone resorption in osteoclasts were analyzed.Results:Compared with the OVX group,HLE exerted bone protective effects by the increase of estradiol(P＜0.05),the improvement of cancellous bone structure,bone mineral density(P＜0.01)and the reduction of serum alkaline phosphatase(ALP),tartrate resistant acid phosphatase(TRAP),bone gla-protein,c-terminal telopeptides of type I collagen(CTX-1)and deoxypyridinoline levels(P＜0.01 for all).In vitro experiment,compared with the control group,HLE at 20 μg/mL promoted the cell proliferation(P＜0.01),and increased the expression of bone morphogenetic protein-2 and osteopontin levels in osteoblasts(both P＜0.05).HLE at 100 μg/mL increased the osteoblastic ALP activities,and HLE at all dose enhanced the extracellular matrix mineralization(both P＜0.01).Furthermore,compared with the control group,HLE at 20 μg/mL and 100 pg/mL inhibited osteoclastic TRAP activity(P＜0.01),and reduced the expression of matrix metalloproteinase-9 and cathepsin K(both P＜0.05).Conclusion:HLE may protect against bone loss,and have potentials in the treatment of osteoporosis.