股密葆含药血清对高浓度地塞米松干预后成骨细胞增殖分化作用的影响

Effects of serum containing Gumibao(股密葆) on the proliferation and differentiation of osteoblast induced by dex-amethasone

目的：观察股密葆含药血清对高浓度地塞米松干预后成骨细胞增殖分化的影响.方法：用多次酶消化法分离出生24 h内新生SD大鼠颅盖骨成骨细胞,P1代细胞进行实验.将不同浓度地塞米松（0、10-8、10-7、10-6、10-5、10-4 mol/L）干预成骨细胞,1周后进行碱性磷酸酶（alkaline phosphatase,ALP）染色,3周后进行矿化结节染色.根据上述实验结果,进一步选择10-5 mol/L地塞米松干预成骨细胞1周,继而改换空白鼠血清及高、中、低浓度股密葆含药血清培养基,培养1周后进行ALP染色,Western-blot检测Ⅰ型胶原及增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）表达,培养2周后进行矿化结节染色.结果：生理浓度地塞米松（10-8 moL/L）可促进成骨细胞ALP的表达及矿化结节产生,而高浓度地塞米松（10-7～10-4 mol/L）可抑制上述结果,股密葆含药血清可逆转高浓度地塞米松对成骨细胞ALP、矿化结节、Ⅰ型胶原及PCNA的抑制作用.结论：高浓度地塞米松可抑制成骨细胞增殖分化,而这种抑制作用可被股密葆含药血清逆转.

Objective：To investigate the effects of serum containing Gumibao （股密葆） on the proliferation and differentiation of osteoblast induced by dexamethasone.Methods：Osteoblasts were extracted from skulls in newly born （within 24 hours） SD rats,and digested with collagenase.The first passage of cells were used for experiments.Cells were cultured in the medium containing different concentrations of dexamethasone （0,10-8,10-7,10-6,10-5,10-4 mol/L）.Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks.Accordingly,following the treatment of 10-5 mol/L dexamethasone for 1 week,cells were cultured in the medium with serum containing G umibao （股密葆）.One week after Cumibao （股密葆） treatment,cells were stained with Alkaline phosphatase and collagen Ⅰ and PCNA were examined by Western-blot.However,the observation of numbers of mineralized nodes with alizarin red stain required one more week.Results：High concentration of dexamethasone could inhibit the expression of PCNA,collagen Ⅰ,alkaline phosphatase and reduce the number of mineralized nodes of osteoblast,while serum containing Gumibao （股密葆） could reverse the inhibition.Conclusion：High concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells,while serum containing Gumibao （股密葆） could reverse the inhibition.