摘要
OBJECTIVE: To investigate the effect of icariin on proliferation of bone marrow mesenchymal stem cells(BMSCs) in Sprague-Dawley(SD) rats.METHODS: 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(Brd U) incorporation were applied to detect the effect of icariin on BMSCs proliferation.Flow cytometry was used to detect proliferation in-dex of BMSCs. The m 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(p GSK-3β)and cyclin D1.RESULTS: Icariin promoted BMSCs proliferation at the concentration of 0.05-2.0 mg/L. The percentage of Brd U positive cells of BMSCs was increased from40.98% 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 m RNA expression, were all increased with icariin treatment. Meanwhile, the phosphorylation level of GSK-3β and cyclin D1 protein expressions were also increased in BMSCs with icariin treatment.CONCLUSION: 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.
OBJECTIVE： To investigate the effect of icariin on proliferation of bone marrow mesenchymal stem cells（BMSCs） in Sprague-Dawley（SD） rats.METHODS： 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（Brd U） incorporation were applied to detect the effect of icariin on BMSCs proliferation.Flow cytometry was used to detect proliferation in-dex of BMSCs. The m 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（p GSK-3β）and cyclin D1.RESULTS： Icariin promoted BMSCs proliferation at the concentration of 0.05-2.0 mg/L. The percentage of Brd U positive cells of BMSCs was increased from40.98% 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 m RNA expression, were all increased with icariin treatment. Meanwhile, the phosphorylation level of GSK-3β and cyclin D1 protein expressions were also increased in BMSCs with icariin treatment.CONCLUSION： 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.
基金
the the National Natural Science Foundation of China(Study on the Bushen Chinese Medicine on the BMSCs Differentiation,No.81173619,81403479)
Natural Science Foundation of the Jiangsu Province of China(Study of Acupuncture on Ischemic Stroke,No.BK20130956)
Specialized Research Fund for the Doctoral Program of Higher Education of China(Epigenetic Mechanism of Acupuncture on Brain Injury,No.20133237120002)
