Effects And Mechanism Of α-zearanol On Culture Osteoblast

Objective:The objective of this research was to clarify the efficacy ofα-Zearalanol to proliferation of osteoblast, NO and expression of c-jun.Methods:Newborn SD rats were sacrificed and soaked in 75% ethanol for 10 min, and then cranium of rats were obtained. The cranium were stripped off all the soft tissue , rinsed and cut to trivial bone block under sterile condition. After digested with 0.25% trypsin for 20 min, the bone block was cultured in culture flask in incubator with 0.1% collagenase and cell culture fluid. Osteoblasts were inoculated in 96-well flat-bottomed plates at a concentration of 10⁵/ml, then the culture plates were randomly divided into 4 groups: control group, estrogen group, 10^-7 mol/Lα-ZAL group and 10^-10 mol/Lα-ZAL group and and treated with correspondent medicine. The follow index was measured:(1) Effects ofα-ZAL on osteoblast apoptotic were detected with flow cytometry.(2) NO density in osteoblast was measured using kit.(3)The expression of c-jun gene in osteoblast was detected by immunohistochemisty.Result:(1) The apoptosis in Estrogen group,α-ZAL10^-7 mol/L group andα-ZAL10^-10 mol/L group was inhibited compared control group.(2) NO density in osteoblast increased inα-ZAL group, and NO inα-ZAL10^-10 mol/L group was higher thanα-ZAL10^-7 mol/L group after 24h, while NO inα-ZAL10^-7 mol/L group was higher thanα-ZAL10^-10 mol/L group(3) The expression of c-jun gene is upregulated byα-ZAL10^-7 mol/L andα-ZAL10^-10. Conclusion:(1)α-ZAL can inhibit apoptosis of osteoblast and improve its proliferation.(2)α-ZAL can regulate NO synthesis of osteoblast, which may be one of the mechanism for the regulation effects ofα-ZAL to osteoblast.(3)α-ZAL can upregulate expression of c-jun gene of osteoblast.