含抗骨增生胶囊血清预处理骨髓间充质干细胞的增殖和成骨分化  

Effects of Kangguzengsheng Capsules-containing serum on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells

作　　者：宋永周[1] 童九辉 刘会玲[1] 马维[1] 李明[1] 

机构地区：[1]河北医科大学第二医院骨科,河北省石家庄市050000

出　　处：《中国组织工程研究》2017年第9期1439-1443,共5页Chinese Journal of Tissue Engineering Research

基　　金：河北省医学科学研究重点课题(20150666)~~

摘　　要：背景:课题组前期研究发现抗骨增生胶囊可促进骨折愈合。目的:观察含抗骨增生胶囊血清对骨髓间充质干细胞增殖和成骨分化的影响。方法:将40只SD大鼠随机分4组,低、中、高剂量组分别灌胃给予抗骨增生胶囊药粉11.6,34.8,104.4 g/kg,空白组灌胃给予等量生理盐水,1次/d,连续灌胃12 d后,腹主动脉采血,分离血清备用。应用全骨髓贴壁培养法分离SD大鼠骨髓间充质干细胞,分4组培养,分别加入含上述4种血清的成骨培养基中诱导培养,培养24,48,72 h,检测细胞增殖;培养7,14 d,检测细胞碱性磷酸酶活性;培养7 d,进行碱性磷酸酶染色;培养14 d,进行茜素红染色。结果与结论:(1)与空白组比较,低剂量组细胞增殖无明显变化,中、高剂量组培养48,72 h的细胞增殖显著升高(P<0.05);(2)与空白组比较,低剂量组细胞碱性磷酸酶活性无明显变化,中、高剂量组培养7,14 d的细胞碱性磷酸酶活性显著升高(P<0.05);(3)低、中、高剂量组均可见碱性磷酸酶及茜素红染色阳性,以高剂量组效果最明显;(4)结果表明,含抗骨增生胶囊血清可促进骨髓间充质干细胞的增殖及成骨分化。BACKGROUND： Our previous studies have found that the Kangguzengsheng Capsules can promote fracture healing. OBJECTIVE： To explore the influence of Kangguzengsheng Capsules-containing serum on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.METHODS：Forty Sprague-Dawley rats were randomly divided into four groups.The capsule powder was resolved into1 mL natural saline and intragastrically administered into rats according to 1.16 g/100 g,3.48 g/100 g,10.44 g/100 g in low,medium and high dose groups.Rats in control group were given equal volume of natural saline.After consecutive administration for 12 days,blood samples from the abdominal aorta were collected and serum samples were isolated and preserved until use.Bone marrow mesenchymal stem cells were isolated and purified by the whole marrow adhesion method,and identified by flow cytometry.Harvested cells were divided into four groups and cultured in the osteogenic culture medium containing different kinds of serum samples as described above.MTT method was adopted to test the cell proliferation at 24,48 and 72 hours of culture.Alkaline phosphatase activity in cells was detected at 7 and 14 days of culture.Alkaline phosphatase staining was performed at 7 days of culture,and alizarin red staining performed at 14 days of culture.RESULTS AND CONCLUSION：Compared with the blank control group,bone marrow mesenchymal stem cells showed no significant changes in the proliferative ability and alkaline phosphatase activity in the low dose group,but these two indices were significantly increased in the high and medium dose groups at 48 and 72 hours of culture or at 7 and 14 days of culture,respectively（P〈0.05）.Cells positive for alkaline phosphatase and alizarin red staining were observed in the low,medium and high dose groups,and the cell staining was most remarkable in the high dose group.To conclude,Kangguzengsheng Capsules-containing serum can promote the proliferation and osteogenic differentiation of bone marrow mesenc

关 键 词：干细胞 间质干细胞 细胞增殖 组织工程 骨髓干细胞 骨髓间充质干细胞 增殖 碱性磷酸酶 含药血清 骨向分化 

分 类 号：R394.2[医药卫生—医学遗传学]