Fu Shuping, Yang Li, Hong Hao, Zhang Ronghua
J Tradit Chin Med. 2016 Jun;36(3):360-8. doi: 10.1016/s0254-6272(16)30050-4.
To investigate the effect of icariin on proliferation of bone marrow mesenchymal stem cells (BMSCs) in Sprague-Dawley (SD) rats.
BMSCs were obtained from SD rat bone marrow with differential time adherent method. Its characteristic was identified through differentiation cell surface antigens and the multi-lineage (osteo/adipo/chondo) differentiation potential. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and 5-Bromo-2-Deoxyuridine (BrdU) incorporation were applied to detect the effect of icariin on BMSCs proliferation. Flow cytometry was used to detect proliferation index of BMSCs. The RNA level and the distribution of β-catenin were evaluated by Real-time Polymerase Chain Reaction (PCR) and Immunofluorescent staining respectively. Western blot was used to detect protein expression levels of β-catenin, glycogen synthase kinase-3 beta (GSK-3β), phospho-glycogen synthase kinase-3 beta (pGSK-3β) and cyclinD1.
Icariin promoted BMSCs proliferation at the concentration of 0.05-2.0 mg/L. The percentage of BrdU positive cells of BMSCs was increased from 40.98% to 70.42%, and the proliferation index value was increased from 8.9% to 17.5% with the treatment of 0.05 mg/L icariin, which significance values were both less than 0.05. Compared with the control group, total and nuclear β-catenin proteins, as well as β-catenin mRNA expression, were all increased with icariin treatment. Meanwhile, the phosphorylation level of GSK-3β and cyclinD1 protein expressions were also increased in BMSCs with icariin treatment.
The findings of the present study demonstrated that low dosage of icariin could promote BMSCs proliferation. The activation of Wnt/β-catenin pathways was involved in this process.
探讨淫羊藿苷对Sprague-Dawley（SD）大鼠骨髓间充质干细胞（BMSCs）增殖的影响。
采用差速贴壁法从SD大鼠骨髓中获取BMSCs。通过分化细胞表面抗原和多向（成骨/成脂/成软骨）分化潜能鉴定其特性。应用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法和5-溴-2-脱氧尿苷（BrdU）掺入法检测淫羊藿苷对BMSCs增殖的影响。流式细胞术检测BMSCs的增殖指数。分别采用实时聚合酶链反应（PCR）和免疫荧光染色评估β-连环蛋白的RNA水平和分布。蛋白质印迹法检测β-连环蛋白、糖原合酶激酶-3β（GSK-3β）、磷酸化糖原合酶激酶-3β（pGSK-3β）和细胞周期蛋白D1的蛋白表达水平。
淫羊藿苷在0.05 - 2.0 mg/L浓度下促进BMSCs增殖。用0.05 mg/L淫羊藿苷处理后，BMSCs的BrdU阳性细胞百分比从40.98%增加到70.42%，增殖指数值从8.9%增加到17.5%，二者的差异均有统计学意义（P均<0.05）。与对照组相比，淫羊藿苷处理后β-连环蛋白的总蛋白和核蛋白以及β-连环蛋白mRNA表达均增加。同时，淫羊藿苷处理的BMSCs中GSK-3β的磷酸化水平和细胞周期蛋白D1蛋白表达也增加。
本研究结果表明低剂量淫羊藿苷可促进BMSCs增殖。Wnt/β-连环蛋白信号通路的激活参与了这一过程。
