黄芪甲苷对大鼠骨骺干细胞增殖及生物学特性的影响

Effect of Astragalosideon Proliferation and Biological Characteristics of Precartilaginous Stem Cells of Rats

目的探究黄芪甲苷对大鼠骨骺干细胞(PSCs)增殖及生物学特性的影响。方法将新生SD大鼠离体培养获取大鼠骨骺干细胞,选择黄芪甲苷对第3代骨骺干细胞进行干预诱导,体外培养,在倒置显微镜下观察细胞生长情况、记录细胞贴壁情况及时间、细胞形态变化及增殖情况。采用免疫组化Ⅱ型胶原单克隆抗体染色方法,检测诱导后细胞的生物学特性,MTT法检测细胞增殖活性。结果经黄芪甲苷干预诱导后,与对照组比较,实验组细胞增殖倍增时间加快(23.20h比30.17h,P<0.05)。第4天、第6天实验组PSCs细胞增殖活性较对照组显著提高,差异有统计学意义[第4天:(0.493±0.138)比(0.420±0.051),第6天:(0.612±0.151)比(0.517±0.153),P<0.05)],实验组Ⅱ型胶原单克隆抗体染色表达阳性优于对照组。结论黄芪甲苷具有促进骨骺干细胞增殖的作用。经黄芪甲苷诱导培养的骨骺干细胞可以正常地分泌特征性基质,并保持骨骺干细胞的生物学特性。

Objective To investigate the effect of astragaloside on proliferation and biological characteristics of precartilaginous stem cellsin Rats.Methods Precartilaginous stem cellswere obtained fromnewborn SD rats and cultured in vitro,then the third generation precartilaginous stem cells were chosen to co-cultured with astragaloside.The growth of cells was observed by inverted microscope and the adhere time,circumstances,morphologic changes and proliferationof cells were recorded.Cell biological characteristics induced by astragaloside was tested with immunohistochemistry Collagen type Ⅱ monoclonal antibody staining,the activity of cells was detected by MMT assay.Results The speed of cell proliferation accelerated in experiment group,with a population doubling time of 23.20 h compared to 30.17 h of control group（P〈0.05）.On Day 4 and Day 6,the cells in experiment group showed higher activity than those in control group,with a significant difference（Day 4：0.493±0.138 vs 0.420±0.051;Day 6：0.612±0.151 vs 0.517±0.153;all P〈0.05）.Collagen typeⅡmonoclonal antibody staining in experiment group and control group were both positive,but the staining was more obvious in experiment group.Conclusion Astragaloside can promote the growth of precartilaginous stem cells.Precartilaginous stem cells which was induced with astragaloside can normallysecrete characteristic matrix and keep the well biological characteristics of precartilaginous stem cells.