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目的:研究川芎嗪(tetramethylpyrazine,TMP)对白血病U937细胞增殖与凋亡的影响,并初步探讨其作用机制。方法:应用CCK-8法检测TMP对U937细胞的增殖抑制,流式细胞仪检测细胞凋亡及周期分布,采用Real-time PCR法检测bcl-2,P27 m RNA表达,Western blot检测bcl-2,caspase-3,cyclin E1,CDK2,P27的表达。结果:川芎嗪呈时间剂量依赖的方式抑制U937细胞的增殖,作用48 h的IC50为160 mg·L-1;并且诱导U937细胞凋亡,阻滞细胞周期于G0/G1期;Real-time PCR及Western blot结果显示川芎嗪使U937细胞中凋亡相关分子bcl-2表达下调,caspase-3上调,周期相关蛋白cyclin E1,CDK2表达下调,P27上调。结论:川芎嗪对白血病U937细胞呈抑制增殖及诱导凋亡的作用,其机制可能是通过影响细胞周期分布,下调bcl-2的表达,最终激活caspase-3,启动凋亡途径,诱导细胞凋亡。
Objective: To study the effect of tetramethylpyrazine (TMP) on the proliferation and apoptosis of leukemia U937 cells and to explore its mechanism. Methods: The inhibitory effect of TMP on the proliferation of U937 cells was detected by CCK-8 assay. The apoptosis and cell cycle distribution were detected by flow cytometry. The expressions of bcl-2 and P27 mRNA were detected by Real-time PCR and bcl-2 , caspase-3, cyclin E1, CDK2, P27 expression. Results: Ligustrazine could inhibit the proliferation of U937 cells in a time-and dose-dependent manner, with an IC50 of 160 mg · L-1 at 48 h and induce apoptosis of U937 cells, arresting cell cycle in G0 / G1 phase. Real-time PCR And Western blot results showed that ligustrazine down-regulated the expression of bcl-2, caspase-3, cyclin E1, CDK2 and P27 in U937 cells. CONCLUSION: Tetramethylpyrazine inhibits the proliferation and induces apoptosis of leukemia U937 cells by down-regulating the expression of bcl-2, affecting the cell cycle distribution, and ultimately activating caspase-3, activating the apoptotic pathway and inducing apoptosis .