不同浓度麝香酮对外源性骨髓间充质干细胞在体内迁移的影响

Effects of Musk Ketone at Different Concentrations on in vivo Migration of Exogenous Rat Bone Marrow Mesenchymal Stem Cells

目的研究不同浓度麝香酮对体外培养大鼠骨髓间充质干细胞(rat bone rnarrow mesenchymal stem cells,rBMSCs)在体内迁移的影响,以期筛选出最佳的治疗剂量。方法 SD大鼠40只,随机分为4组,每组10只,采用牙科钻营造颅骨缺损的大鼠模型;利用贴壁筛选法提取大鼠骨髓间充质干细胞,传至第3代,并加入含有终浓度为10μmol/LBrdU进行标记,待细胞融合至于80%时,利用胰酶消化,配置成细胞密度为1×109cells/L细胞混悬液,然后通过尾静脉注入颅骨缺损的大鼠体内,回植后分别采用高(16.8μL/100g)、中(8.4μL/100g)、低(4.2μL/100g)剂量的麝香酮灌服实验大鼠,空白组以灌服等体积生理盐水作为对照。14d后取出颅骨,固定,脱钙,做免疫组化后,荧光显微镜下,计数BrdU阳性细胞。结果利用免疫组化处理后,进行灰度分析,麝香酮低、中剂量组与空白组比较,差异有统计学意义(P<0.01),麝香酮高剂量组与空白组比较,差异无统计学意义(P>0.05)。结论麝香酮具有促进外源性干细胞在体内的迁移作用,以中、低浓度麝香酮效果较好。

Objective To study the effects of musk ketone at different concentrations on in vivo migration of exogenous rat bone marrow mesenchymal stem cells （rBMSCs）, thus screening out the optimal therapeutic dose. Methods Forty SD rats were randomly divided into 4 groups, 10 in each group. The rat model of skull de- fect was established using dental surgery. The primary rBMSCs were cultured by adherence screening method. The third passage cells were labeled by 10 μmol/L BrdU, and the labeled cells were injected into skull defect rats from the tail vein. Rats were administered with musk ketone at high, moderate and low concentration, respective- ly by gastrogavage, while equal volume of normal saline was administered to those in the blank control group by gastrogavage. Their skulls were taken out 14 days later, fixed, and decalcified. BrdU positive cells were counted under fluorescence microscope. Results After immunohistochemical processing, the gray scale analysis was preformed. There was statistical difference in the BrdU positive cell number between the blank control group and the low and moderate concentration musk ketone groups （ P 〈 0. 01 ）. There was no statistical difference in the BrdU positive cell number between the blank control group and the high concentration musk ketone group （P〉0.05）. Conclusion Musk ketone could accelerate the in vivo migration of exogenous stem cells, with the optimal effects obtained at moderate and low concentrations.