蛇床子素对体外培养大鼠股骨组织吸收的抑制作用

Inhibition of osthole for resorption of rats femur tissue in vitro

目的 :研究蛇床子素(Osthole,OS)对体外培养股骨组织(骨干和骨骺端)吸收活性的影响。方法 :取(80±5)g雄性SD大鼠6只,体外分离培养大鼠股骨组织的骨干和骨骺端,随机分为对照组、雌二醇组(estradiol,E)和蛇床子素处理组。同时在股骨组织分离培养48 h后采用终浓度为1×10-5mol/L的蛇床子素和1×10-8mol/L雌二醇对体外培养骨干和骨骺端进行处理;分别在药物处理后的第3、6、9、12天后测定抗酒石酸酸性磷酸酶(Tartrate-resistant acid phosphatase,Str ACP)活性、培养基中葡萄糖(Glucose,Glu)、乳酸(Lactic acid,La)的含量、Real-Time RT-PCR Str ACP、集落刺激因子(Macrophage colony stimulating factor,MCSF)、组织激酶K(Cathepsin K,CTSK)m RNA的表达水平。结果:1×10-5mol/L蛇床子素组碱性磷酸酶(ALP)活性为2226,1×10-8 mol/L雌二醇组ALP活性为2498;与对照组比较1×10-5mol/L蛇床子素和1×10-8mol/L雌二醇组在加药处理后的第6、9和12天显著抑制Str ACP酶活性(P<0.05);1×10-5mol/L蛇床子素处理体外培养大鼠骨骺端和骨干组织的第3、6、9天后显著增加培养基中乳酸含量(P<0.05),并显著减少培养基中葡萄糖的含量(P<0.05)。与对照组比较,1×10-5mol/L的蛇床子素处理体外培养大鼠股骨组织后的第6和9天蛇床子素能显著抑制Str ACP,MCSF,CTSK m RNA的表达水平(P<0.05)。结论 :蛇床子素抑制体外培养大鼠股骨组织的骨骺端和骨干吸收活性同时可提高营养代谢水平。

Objective：To investigate osthole effect on femoral tissue resorption activity of rat in vitro. Methods：Six SD rats weighted（80±5） g were used to isolate and culture femoral tissue（diaphyses and metaphysis） in vitro. The cultured tissue were devided into control group,estradiol group and osthole group. The femoral tissue was treated with final concentration of 1×10-5mol / L osthole and 1 ×10-8mol / L estradiol culture in vitro at 48 hours after cultured. Tartrate resistant acid phosphatase（Str ACP） activity,glucose and Lactic acid content,Str ACP,MCSF（Macrophage colony stimulating factor） and CTSK（Cathepsin K） m RNA was detected by Real-Time RT-PCR were detected. Results：Concetration of Alkaline phosphatase activity were 2226 and 2498 in 1×10-5mol / L osthole and 1×10-8mol / L estradiol respectively. As compared with control group,the activity of Str ACP of 1×10-5mol / L osthole and 1×10-8mol / L estradiol were inhibited at 6,9,12 days（P〈0.05）;under treatment of in 1×10-5mol / L osthole,the content of Lactic acid were increased and the content of glucose were decreased at 3,6,9days（P〈0.05）;Str ACP,MCSF and CTSK m RNA expression level were inhibited at 6,9 days（P〈0.05）. Conclusion：Osthole can inhibit bone resorption and raise the level of nutrition metabolism of femurs tissue.