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目的:研究柠檬醛对人白血病K562细胞株增殖和凋亡的影响,并对其作用机制进行初步探讨。方法:采用本实验改良的MTT比色法和Annexin-V/PI双染色流式细胞术测定柠檬醛对K562细胞的增殖抑制率和凋亡诱导率;采用基于分光比色原理的各种试剂盒定量分析柠檬醛胁迫下K562细胞内丙二醛(MDA)和还原型谷胱甘肽(GSH)含量及谷胱甘肽-S转移酶(GST)活性。结果:K562细胞经柠檬醛作用后,6.9~444 mg.L-1柠檬醛抑制K562细胞增殖呈时效性及量效性;111,222mg.L-1柠檬醛可诱导K562细胞凋亡,前者主要表现为诱导早期凋亡活性(4 h:4.3%,24 h:36.6%),后者则主要表现为诱导晚期凋亡活性(4 h:23.9%,24 h:52.4%);柠檬醛作用于K562细胞引起细胞氧化应激的改变,111 mg.L-1柠檬醛作用3 h使K562细胞MDA水平上升为对照组的3.29倍,而GST活性几乎完全抑制。不同浓度柠檬醛作用均引起细胞GSH水平下降。结论:柠檬醛胁迫可有效抑制K562细胞增殖并诱导其凋亡,细胞内GSH含量下降、柠檬醛高蓄积及ROS水平上升可能是其抑制作用的重要分子机制,而ROS途径可能是柠檬醛诱导细胞凋亡机制之一。
Objective: To study the effects of citral on the proliferation and apoptosis of human leukemia K562 cell line, and to explore its mechanism. Methods: MTT colorimetric assay and Annexin-V / PI double staining flow cytometry were used to determine the inhibition of cell growth and apoptosis induced by citral on K562 cells. Various kits based on spectrophotometric colorimetry The contents of malondialdehyde (MDA), reduced glutathione (GSH) and glutathione S-transferase (GST) in K562 cells under citral stress were quantitatively analyzed. Results: After citral, K562 cells inhibited the proliferation of K562 cells with the dose of 6.9 ~ 444 mg.L-1 citral for a time-dependent and dose-dependent manner. 111,222 mg.L-1 citral could induce K562 cell apoptosis, (4 h: 4.3%, 24 h: 36.6%), while the latter mainly induced late apoptotic activity (4 h: 23.9%, 24 h: 52.4%). Citral acted on K562 Cell oxidative stress caused by cell changes, 111 mg.L-1 citral 3 h K562 cell MDA levels increased 3.29 times the control group, while the GST activity almost completely inhibited. Different concentrations of citral caused cell GSH levels decreased. CONCLUSION: Citral can effectively inhibit the proliferation and induce the apoptosis of K562 cells, the decrease of intracellular GSH, the high accumulation of citral and the increase of ROS levels may be the important molecular mechanisms of its inhibition. ROS pathway may be citral-induced cells One of the mechanisms of apoptosis.