葡萄糖对人骨髓间充质干细胞增殖的影响及调控机制

摘要

Objective To investigate the effects of glucose on human mesenchymal stem cells(hMSCs)proliferation and the possible molecular mechanism. Methods hMSC TERT cells were grown either in 5. 6 or 11. 0 or 25 mmol/L glucose for 14 and 28 days. The proliferation rate was measured by the MTT assay. Cells were analyzed by using cDNA microarray and gene ex pression profiles were obtained. RT PCR was done to validate the expression of TXNIP mRNA in response to glucose. Results The proliferation rate of hMSC TERT cells was decreased with the increased concentration of glucose and prolonged incubation time. The TXNIP gene expression in hMSC TERT cells was up regulated with the increased levels of glucose and persisted through the duration of the conditions of hyperglycemia. The expression of TXNIP was significantly increased in 5. 6 mmol/L glucose group as compared with 11. 0 or 25. 0 mmol/L glucose group(P<0. 01). There was no significant difference in the ex pression of TXNIP between 11. 0 and 25. 0 mmol/L glucose group(PX). 05). In 5. 6 mmol/L glucose group,the expression of TXNIP mRNA was significantly increased after incubation for 28 days as compared with that after incubation for 14 days(P< 0. 01). There was no significant difference in the expression of TXNIP between 11. 0 and 25. 0 mmol/L glucose groups follow ing 14 or 28 days incubation(P>0. 05). Conclusion The proliferation capacity of hMSC TERT cells is reduced under a high glucose condition,which is associated with the up regulation of TXNIP mRNA.%目的 观察不同浓度葡萄糖对人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs)增殖的影响,并初步探讨其可能的基因调控机制.方法 hMSC-TERT细胞在不同浓度葡萄糖(5.6、11.0、25.0 mmol/L)培养液中,分别培养14、28 d后,用MTT法测定各组hMSC-TERT细胞的增殖率,基因芯片检测hMSC-TERT细胞在高浓度葡萄糖(25.0 mmol/L)与生理浓度葡萄糖(5.6 mmol/L)作用下基因谱的差异性表达,并用RT-PCR扩增技术进一步证实不同浓度葡萄糖作用下的hMSC-TERT细胞硫氧还蛋白相关蛋白(TXNIP)mRNA表达的差异.结果 细胞增殖测定结果显示随着葡萄糖浓度的增加以及培养时间的延长,hMSC-TERT细胞的增殖率下降.同时基因芯片测定筛选出表达差异的基因有87条,其中12条表达上调,75条表达下调.在表达上调的基因中,TXNIP基因表达增加最为显著.RT-PCR结果也进一步证实在相同培养时间下,与5.6 mmol/L葡萄糖浓度组相比,11.0 mmol/L和25.0 mmol/L 葡萄糖浓度组TXNIP mRNA表达均明显增加(均P＜0.01);25.0 mmol/L 组较11.0 mmol/L 组TXNIP mRNA表达增加,但差异无统计学意义(P＞0.05).在5.6 mmol/L葡萄糖浓度组,培养28 d时TXNIP mRNA水平较培养14 d时明显增加(P＜0.01);在11.0 mmol/L 和25.0 mmol/L 葡萄糖浓度组间,培养至14 d与28 d后,TXNIP mRNA水平的改变差异均无统计学意义(P＞0.05).结论 高糖环境中hMSC-TERT细胞的增殖能力下降,TXNIP mRNA的表达增加调节hMSC-TERT细胞增殖能力的改变.