Wang Xiao-jing, Yang Gui-cun, Chen Hong-xia, Zhang Ping, Xu You-hua
Zhongguo Zhong Yao Za Zhi. 2015 Jun;40(11):2186-90.
To study the proliferation and apoptosis of tetramethylpyrazine (TMP) on leukemic U937 cells and its possible mechanism.
The inhibitory effect of TMP on the proliferation of U937 cells was detected by CCK-8 assay. The cell apoptosis and cycle distribution were examined by the flow cytometry. The mRNA expressions of bcl-2 and P27 were determined by the Real-time PCR. Western blot was carried out to detect bcl-2, caspase-3, cyclin E1, CDK2 and P27 expressions.
TMP inhibited the proliferation of U937 cells in a dose-and-time dependent manner, with IC50 value of 160 mg x L(-1) at 48 h. In addition, TMP could induce the apoptosis of U937 cells and block the cell cycle in G0/G1 phase. According to the results of Real-time PCR and Western blot, TMP could down-regulate the expression of apoptosis-related molecule bcl-2, cycle-related protein cyclin E1 and CDK2 and up-regulate caspase-3 and P27.
TMP shows the effects in inhibiting the proliferation of leukemic U937 cells and inducing the apoptosis. Its mechanism may be related to the impacts on the cell cycle distribution, down-regulation of the bcl-2 expression, which finally activates caspase-3, starts the apoptosis path and causes the cell apoptosis.
研究川芎嗪(TMP)对白血病U937细胞增殖和凋亡的影响及其可能机制。
采用CCK-8法检测TMP对U937细胞增殖的抑制作用。通过流式细胞术检测细胞凋亡和细胞周期分布。采用实时荧光定量PCR法检测bcl-2和P27的mRNA表达。采用蛋白质免疫印迹法检测bcl-2、caspase-3、细胞周期蛋白E1、细胞周期蛋白依赖性激酶2(CDK2)和P27的表达。
TMP对U937细胞增殖的抑制作用呈剂量和时间依赖性,48 h时IC50值为160 mg·L-1。此外,TMP可诱导U937细胞凋亡,并将细胞周期阻滞于G0/G1期。实时荧光定量PCR和蛋白质免疫印迹结果显示,TMP可下调凋亡相关分子bcl-2、细胞周期相关蛋白细胞周期蛋白E1和CDK2的表达,上调caspase-3和P27的表达。
TMP具有抑制白血病U937细胞增殖和诱导凋亡的作用。其机制可能与影响细胞周期分布、下调bcl-2表达,最终激活caspase-3、启动凋亡途径并导致细胞凋亡有关。