L型钙通道对大鼠骨髓间充质干细胞增殖及细胞周期影响的研究

目的 研究L型钙通道对骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)增殖及细胞周期的影响。方法 SD大鼠BMSCs原代培养、传代;RT-PCR检测BMSCs钙通道基因的表达;取第3代BMSCs,分为2组,实验组加入L型钙通道阻断剂硝苯地平(nifedipine,Nif),对照组不加药。MTT法测量其吸光度(A)值、流式细胞仪观察细胞周期的变化。结果 RT-PCR结果显示,L型钙通道alC、CCHL2a亚基的mRNA呈高表达;L型钙通道阻断后,从BMSCs增殖第3天开始,实验组与对照组A值比较,组间差异均具有统计学意义;经Nif作用后,实验组S、S+G2M期细胞比例减少,GO/G1期细胞比例较对照组高,组间差异有统计学意义。结论 阻断L型钙通道可抑制BMSCs的DNA合成进而抑制细胞增殖。

Effects after blocked L-type calcium channels on rat bone mesenchymal stem cells

Objective To investigate the effects of L-type calcium channels on rat bone mesenchymal stem cells. Methods Marrow mesenehymal stem cells were separated and cultured in vitro. RT-PCR was used to detect the mRNA expression of alC, CCHL2a . The L-type calcium channel blocker, nifedipin, was used to block the L-type calcium channels. Methyl thiazol tetrazolium assay and flow cy- tometry were applied to measure the cellular proliferation and cellular cycle. Results A high expression of mRNA in alC was detected by RT-PCR. The cells in treatment group proliferated more slowly than those in the control group after blocking L-type calcium channels. There was significant difference of two groups after continuous culture for 3 days. After 72 hours stimulation, the percentage of phase S, S ＋ G2M was decreased, compared with the control group, and the percentage of phase GO/G1 increased, compared with the control group . There were significant differences between the two groups. Conclutions L-type calcium channels may be involved in the proliferation of marrow mesenchymal stem cells.