携带重组腺病毒 BMP-2的巨噬细胞对 C3H10细胞成骨分化的影响

目的：探讨携带 BMP-2（Bone Morphogenetic protine-2）腺病毒的巨噬细胞对小鼠 C3H10细胞的增殖和成骨分化的作用。方法：感染重组腺病毒 BMP-2的巨噬细胞与小鼠 C3H10细胞共培养。MTT 方法检测共培养体系中 C3H10细胞的增殖能力；共培养7 d 对 C3H10细胞进行 ALP 染色和活性检测；细胞培养至21 d，进行标茜素红染色检测矿化结节；定量 PCR 实验验证过表达腺病毒载体 BMP-2有效性并且检测 Runx2的表达；Western blot检测细胞中磷酸化 Smad1/5/8蛋白表达。结果：与对照组相比，巨细胞过表达 BMP-2与 C3H10细胞共培养后，可以促进 C3H10细胞增殖；ALP 染色程度和活性上升；矿化结节增多；定量 PCR 结果显示 Ruxn2 RNA 表达增加， Western blot 结果显示 Ruxn2蛋白表达上升。结论：过表达 BMP-2的巨噬细胞可以促进小鼠 C3H10细胞的增殖，并且可能通过 BMPs 经典途径促进 C3H10细胞的成骨分化。

Macrophage carrying BMP-2 promote C3H10 cell osteogenic differentiation

Objective To investigate effects of macrophage carrying BMP-2 on C3H10 cell osteogenic differentiation. Methods macrophage cell which was infected recombinant adenovirus BMP-2 co-culture with C3H10 cells. MTT was used to detect the proliferation capacity of C3H10 cell. C3H10 cell was performed ALP histochemical stain and activity detection at 5 and 7 days. mineralized nodules were exa mined by alizarin red staining, after 21 days. Real-time quantitative PCR was used to confirm the recombinant adenovirus BMP-2 availability and analyze Runx2 expression level. p-smad1/5/8 was analyzed by Western blot. Results Compared with control group, overexpression of BMP-2 in macrophage cell in co-culture system, the proliferation of C3H10 cells was promoted, ALP activity was significantly stimulated,and matrix mineralization increased.Re-al-time quantitative PCR results showed that the mRNA level of Runx2 was increased, and Western blot results show that the Rxun2 was increased in BMP-2 overexpression group. Conclusion Proliferation and osteogenic differentiation of C3H10 cell was promoted in co-culture system of over-expressing BMP-2 in macrophage cell, and the mechanism is potential achieved via stimulating typical BMP signal pathway.