从细胞水平研究龟鹿二仙含药血清在治疗骨质疏松中的作用

从细胞水平研究龟鹿二仙含药血清在治疗骨质疏松中的作用.(1)用酶消化法获得新生大鼠的成骨细胞.碱性磷酸酶(ALP)染色进行鉴定,通过对成骨细胞增殖、胰岛素样生长因子(IGF-1)分泌的检测来考察成骨细胞功能.(2)用1,25(OH)2D3诱导骨髓单核细胞获得破骨细胞,抗酒石酸酸性磷酸酶(TRAP)染色鉴定破骨细胞.电镜扫描观察破骨细胞形成的骨凹陷情况;同时检测TRAP( )细胞数和破骨细胞TRAP活性.(3)用血清药理学方法制备龟鹿二仙含药血清,分别加入不同剂量组10%含药血清进行干预,观察该中药血清对上述成骨细胞和破骨细胞功能指标的影响.龟鹿二仙高剂量组血清可促进成骨细胞增殖,增加IGF-1的分泌量.而中剂量组血清可明显抑制骨髓细胞向破骨细胞的转化,抑制骨凹陷形成,降低细胞内TRAP的活性.本研究表明龟鹿二仙具有显著的抗骨质疏松作用.

Effects of Serum Containing Oral Liquid of Guilu-Erxian on the Therapy of Osteoporosis at the Cellular Level

This study sought to assess the effects of serum containing Guilu-Erxian (GLEX) on treating osteoporosis at the cellular level. First, Enzyme-digestion was used to obtain osteoblasts from newborn SD rats. Alkaline phosphatase staining was used to identify osteoblasts. The proliferation and the secretion of Insulin Like Growth Factor-1 (IGF-1) of osteoblasts were examined. Second, osteoclasts were generated by inducing bone marrow cells of SD male rat (6-9 weeks) with 1,25(OH)2D3. The osteoclasts were identified through tartrate-resistant acid phosphatase (TRAP) staining. Bone pits of osteoclasts were observed through Electron Scanning Microscope. At the same time, TRAP(+) cells with more than two nuclei and TRAP activity of osteoclasts were examined. Last, serum containing Guilu-Erxian was made through serum pharmacology. And we observed the effects of serum containing Guilu-Erxian on the function of osteoblasts and osteoclasts. We found that high doses of serum containing Guilu-Erxian had obvious effects on stimulating the multiplication and capability of secreting IGF-1 of osteoblasts cultured in vitro. Middle doses obviously inhibited the formation of osteoclasts and bone pits; TRAP activity of osteoclasts was also lowered clearly. This study indicated that Guilu-Erxian has positive effects on treating osteoporosis.