湖南中医药大学学报2023，Vol.43Issue(12)：2163-2170,8.DOI:10.3969/j.issn.1674-070X.2023.12.004
桃红四物汤诱导间充质干细胞动员促进骨折修复的机制
Mechanism of Taohong Siwu Decoction inducing mesenchymal stem cell mobilization to promote fracture repair
摘要
Abstract
Objective To study the effect of Taohong Siwu Decoction(THSWD)on bone marrow mesenchymal stem cell mobilization in rats with early fracture.Methods A total of 24 SPF male SD rats were randomly assigned into normal,fracture,and drug groups respectively,with eight rats in each group.Except for the normal group,the other groups received the surgery of a right femoral shaft fracture.On the first day after the operation,the drug group was given 20 g/kg of THSWD for intragastric administration,while the normal group and the fracture group were given the same amount of distilled water.All rats were administered intragastrically in the morning and evening for five days.The number of CD45-CD90+CD29+ monocytes in peripheral blood was determined by flow cytometry.The surface markers were identified also by flow cytometry,cell morphology was observed under an inverted microscope,osteogenic and adipogenic differentiation were observed by alizarin red staining and oil red O staining,respectively,to identify the origin of peripheral blood MSCs.The expression of cytokines in the early stage of fracture was detected by protein chips.Results The number of CD45-CD90+CD29+ monocytes in peripheral blood in fracture rats was significantly higher than that of normal rats(P<0.05).The number of MSCs n peripheral blood was further elevated in the durg group(P<0.01).Compared with the fracture group,the number of MSCs in peripheral blood of drug group increased(P<0.05).Flow cytometry analysis revealed that the proportions of CD45-,CD90+,CD29+ cells and CD45-CD90+CD29+ cells in bone marrow-derived cells of THSWD-interfered rats were 90.94%,98.57%,92.27%,and 94.15%.The proportions of CD45-,CD90+,CD29+ cells,and CD45-CD90+CD29+ cells in peripheral blood-derived cells of THSWD-interfered rats were 90.59%,98.21%,92.03%,and 93.52%.By obs erving the cell morphology under the microscope,it was found that peripheral blood MSCs had a significantly longer growth cycle and a significantly reduced number of cell clones compared with bone marrow MSCs.There is no significant difference in cell morphology;after the induction of osteogenic and adipogenic differentiation,alizarin red and oil red O staining revealed that both peripheral blood MSCs and bone marrow MSCs could turn towards osteogenic and adipogenic differentiation.Compared with the normal group,the expressions of cytokine-induced neutrophil chemoattractant-1(CINC-1)and macrophage inflammatory protein-1α(MIP-1α)were higher in the peripheral blood in the fracture group(P<0.05).Compared with the fracture group,the expressions of CINC-1,fractalkine(FKN),L-selectin,vascular endothelial growth factor(VEGF)proteins increased(P<0.05),and the expression of interleukin-1α(IL-1α)protein decreased(P<0.05)in peripheral blood of drug group.Conclusion Early intervention of THSWD,the representative formula to promote blood circulation and remove blood stasis,can signific antly induce the mobilization of bone marrow MSCs in rats,and its molecular mechanism may be related to the up-regulation of CINC-1,FKN,L-selectin,and VEGF,and the down-regulation of IL-1 in peripheral blood.关键词
桃红四物汤/骨折/骨折修复/间充质干细胞/动员Key words
Taohong Siwu Decoction/fracture/fracture repair/mesenchymal stem cells/mobilization分类
医药卫生引用本文复制引用
曾朋,陆小龙,徐无忌,向黎黎,李汪洋,熊辉..桃红四物汤诱导间充质干细胞动员促进骨折修复的机制[J].湖南中医药大学学报,2023,43(12):2163-2170,8.基金项目
国家自然科学基金面上项目(81874478) （81874478）
湖南省中医药管理局一般项目(2022-04033025) （2022-04033025）
湖南省教育厅青年项目(22B0369). （22B0369）
