世界中医药

目的：观察接骨丹对新西兰乳兔成骨细胞增殖的影响，探讨接骨丹促进骨折愈合的机制。方法：普通级新西兰大白乳兔，耳缘静脉空气栓塞处死，无菌条件下取颅盖骨，Ⅰ型胶原酶消化法获取成骨细胞并进行体外培养、鉴定及传代。将鉴定后的第二代成骨细胞随机分为正常对照组及实验组,正常对照组应用正常兔血清培养,实验组应用接骨丹含药血清培养,MTT法观测2组细胞的生长曲线；不同血清干预48 h后，倒置显微镜观测接骨丹对成骨细胞碱性磷酸酶及Ⅰ型胶原表达的影响，Elisa法观测接骨丹对成骨细胞培养上清液中骨钙素表达的影响。结果：Ⅰ型胶原酶消化法成功获取并建立成骨细胞体外培养体系，MTT观测显示实验组能够显著促进成骨细胞的增值，倒置显微镜观测实验组中碱性磷酸酶及Ⅰ型胶原的表达要优于正常对照组，并能够促进成骨细胞分泌骨钙素。结论：接骨丹能够促进骨折的愈合，其机制可能与促进成骨细胞中碱性磷酸酶、Ⅰ型胶原及骨钙素的表达进而促进成骨细胞增殖和分化有关。

English Summary:

To observe the effect of Bone-Joining Elixir on New Zealand newborn rabbit osteoblast proliferation, and explore the promoting fracture healing mechanism of Bone-Joining Elixir. Methods: Conventional grade New Zealand big and white newborn rabbits were put to death by ear venous air embolism, the craniums were taken down in aseptic conditions, and the osteoblasts were gained by collagenase type Ⅰ digestion method for in vitro culture, identification and passages. After identification, the second generation of osteoblasts was randomly divided into normal control group and experimental group. And the normal control group was cultured in normal rabbit serum, the experimental group was cultured in Bone-Joining Elixir medicated serum, MTT method was taken to observe the cell growth curves of the two groups. After the intervention for 48 h, the two serums were observed to achieve the influence of Bone-Joining Elixir on the expression of osteoblast alkaline phosphatase and collagen typeⅠ under the inverted microscope, and to achieve the influence of Bone-Joining Elixir on the expression of osteocalcin in the supernate in the osteoblast culture by Elisa method.

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