论文部分内容阅读
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。
Objective: To investigate the effects of macrophages carrying BMP-2 adenovirus on the proliferation and osteogenic differentiation of mouse C3H10 cells. Methods: Macrophages infected with recombinant adenovirus BMP-2 were co-cultured with mouse C3H10 cells. The proliferation of C3H10 cells in co-culture system was detected by MTT assay. ALP staining and activity detection of C3H10 cells were performed 7 days after co-culture. Cell culture was performed for 21 days. Alizarin red staining was used to detect mineralized nodules. The expression of adenovirus vector BMP-2 was detected and the expression of Runx2 was detected. The protein expression of phosphorylated Smad1 / 5/8 was detected by Western blot. Results: Compared with the control group, the co-culture of giant cell overexpression BMP-2 and C3H10 cells could promote the proliferation of C3H10 cells; the level and activity of ALP staining increased; the number of mineralized nodules increased; the quantitative PCR results showed that Ruxn2 RNA expression increased, Western blot results showed that Ruxn2 protein expression increased. Conclusion: Macrophages overexpressing BMP-2 can promote the proliferation of mouse C3H10 cells and promote the osteogenic differentiation of C3H10 cells through the classical pathway of BMPs.