麝香对颅骨骨缺损模型大鼠单核细胞趋化蛋白1表达的影响

摘要

目的 观察不同剂量麝香对颅骨骨缺损模型大鼠骨缺损处单核细胞趋化蛋白1(MCP-1)表达的影响,探讨麝香促进外源性骨髓间充质干细胞(rBMSCs)向骨缺损处迁移的机制.方法 采用贴壁筛选法培养rBMSCs,用流式细胞仪鉴定.大鼠颅骨骨缺损模型建立后回植rBMSCs.将模型组大鼠完全随机分为中药高、中、低剂量组及空白组,中药高、中、低剂量组灌服不同剂量的麝香,空白组灌服生理盐水.14 d 后处死大鼠,取出缺损部位的颅骨,并对其脱钙、制作切片,免疫组化染色,光学显微镜下观察,每个切片随机选取3 个视野拍照,将所得照片用IPP6.0 处理计算IOD 值,再进行统计分析.结果 筛选纯化的rBMSCs 呈均一的成纤维细胞样,贴壁生长,以长梭形为主,漩涡状盘旋排列,表型鉴定结果为CD45 阴性表达,CD44、CD90 表达阳性,rBMSCs 经诱导后可向成脂、成骨分化,麝香能促进颅骨骨缺损模型大鼠骨缺损处MCP-1 表达,中药各组与空白组比较差异有统计学意义(P ＜0.01),以低浓度效果最佳(P ＜0.05).结论 麝香促进rBMSCs 在大鼠体内向损伤部位迁移的机制与其促进骨缺损处MCP-1 表达有关.%Objective To observe the effects of different doses of musk on MCP-1 expression of skull bone defect model rats and explore its mechanism of promoting exogenous rat bone mesenchymal stem cells (rBMSCs) migrate to the bone defect. Methods Adherence screening method was emplyed to cultivate BMSCs, flow cytometry to identify rBMSCs. The BMSCs were replanted after establishment of skull bone defect model in rats. Model rats were randomly divided into high-, medium-, low-dose group and blank control group, and treated with high-, medium- and low- dose of musk and normal saline respectively. After 14 days, rats were killed and removed the defect parts of skull, decalcified and sliced, and treated with immunohistochemical staining to observe under the microscope, randomly selected three horizons photographed in each slice. The resulting photos IOD was calculated with IPP6.0 processing. Results The purified rBMSCs was homogeneous fibroblast-like, adherent growth, long spindle, swirling circling arrangement. Phenotypic identification result was CD45 negative expression and CD44, CD90 positive expression. The rBMSCs differentiated to adipogenic and osteogenic after induced. Musk promoted the expression of MCP-1 at the bone defect of skull bone defect model rat, and there was significant difference between blank control group and different dose groups (P<0.01), in which the low dose was the best (P<0.05). Conclusion The mechanism of musk promoting exogenous rBMSCs migrating to injured parts is related to promoting the MCP-1 expression at the bone defect of skull bone defect model rat.