【摘要】 目的探讨核因子(NF)-κB在人类骨髓间充质干细胞增殖调节中的作用。方法体外分离培养人骨髓间充质干细胞后:①蛋白免疫印迹法(Western-blot)观察脂多糖(LPS)作用下NF-κB活性亚型p65及其活性形式磷酸化p65的表达水平随时间变化的情况;②Western-blot法观察NF-κB活性抑制剂PDTC对磷酸化p65表达水平影响;③Western-blot法检测PDTC与LPS单独及共同作用下,细胞增殖标志物增殖细胞核抗原(PCNA)表达水平的变化;④MTT法检测PDTC与LPS作用下细胞增殖水平的变化。结果 LPS可显著上调p65及其磷酸化状态的表达水平,上调作用可被PDTC部分地抑制;NF-κB上调后可提高细胞增殖蛋白PCNA的表达水平,并促进细胞增殖。结论初步证实了NF-κB参与人骨髓间充质干细胞增殖的调节。更多还原

【Abstract】 Objective To investigate the role of NF-κB in regulating proliferation of human mesenchymal stem cells(hMSCs).Methods hMSCs were successfully separated and cultured in vitro.Western blotting(WB) was performed to detect the changes in the expression levels of p65,the active sub-unit of NF-κB,along with the time,as well as its phosphated form(p-p65) in hMSCs treated with LPS.The p-p65 expression level was detected by WB in hMSCs treated with PDTC and LPS alone or in combination.The expression of PCNA was detected by using WB in hMSCs treated with PDTC and LPS alone or in combination.The proliferation level was measured by using MTT assay in hMSCs treated with PDTC and LPS alone or in combination.Results The expression levels of both p65 and p-p65 were up-regulated significantly by LPS,and the augmentation was pulled back partially by PDTC;The expression of PCNA could be up-regulated by activated NF-κB,followed by the promoted proliferation of hMSCs.Conclusion It has been proved largely that NF-κB is involved in the regulation of hMSCs proliferation.更多还原