Lou Zheng, Wei Qing-Qing, Wang Da-Wei, Gu Huan-Peng
The 2nd Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou 310053, China.
School of Stomatology, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Zhongguo Zhong Yao Za Zhi. 2018 Jun;43(12):2612-2617. doi: 10.19540/j.cnki.cjcmm.20180320.002.
Allicin is one of the main bioactive substances in garlic, with antibacterial, hypolipidemic and other pharmacological effects. In this study, apoptosis-related indicators were detected to explore the molecular mechanism of allicin on KG-1 cell proliferation inhibition. The apoptosis rate of KG-1 cells induced by allicin was detected by flow cytometry. The effect of allicin on the expressions of Bax, Bcl-2, survivin and ERK mRNA in KG-1 cells was detected by RT-qPCR. Western blot was used to detect the expressions of caspase 3, cleaved caspase 3, ERK1/2, p-ERK1/2 and survivin protein in KG-1 cells. According to the findings, compared with the control group, allicin could significantly inhibit the proliferation activity of KG-1 cells in a concentration-dependent and time-dependent manner. Flow cytometry showed that allicin could induce the apoptosis of KG-1 cells, which was mainly late apoptosis. The results of RT-qPCR showed that the expressions of Bax mRNA, Bcl-2, survivin and ERK mRNA in KG-1 cells increased after treatment with allicin. The results of Western-blot showed that after KG-1 cells were treated with allicin, the expressions of caspase 3 and its active form cleaved caspase 3 increased, the expressions of survivin, ERK1/2 and its active form p-ERK1/2 were decreased, of which p-ERK1/2 was down-regulated in a dose-dependent manner. The above results suggest that allicin inhibited the proliferation of KG-1 cells primarily by inducing late apoptosis; the execution of apoptosis involved cleaved caspase 3; the induction of apoptosis involved the protein expression, the decrease of ERK1/2andexpression of survivin and the dose-dependent decrease of p-ERK1/2; the mRNA expression involved the increase of Bax, and the down-regulation of survivin, Bcl-2 and ERK1/2.
大蒜素是大蒜中的主要生物活性物质之一，具有抗菌、降血脂等药理作用。本研究通过检测凋亡相关指标，探讨大蒜素抑制KG-1细胞增殖的分子机制。采用流式细胞术检测大蒜素诱导KG-1细胞的凋亡率。运用RT-qPCR检测大蒜素对KG-1细胞中Bax、Bcl-2、survivin和ERK mRNA表达的影响。采用蛋白质免疫印迹法检测KG-1细胞中caspase 3、裂解型caspase 3、ERK1/2、磷酸化ERK1/2（p-ERK1/2）和survivin蛋白的表达。结果显示，与对照组相比，大蒜素能以浓度和时间依赖性方式显著抑制KG-1细胞的增殖活性。流式细胞术表明，大蒜素可诱导KG-1细胞凋亡，且主要为晚期凋亡。RT-qPCR结果显示，大蒜素处理后，KG-1细胞中Bax mRNA、Bcl-2、survivin和ERK mRNA的表达增加。蛋白质免疫印迹结果表明，大蒜素处理KG-1细胞后，caspase 3及其活性形式裂解型caspase 3的表达增加，survivin、ERK1/2及其活性形式p-ERK1/2的表达降低，其中p-ERK1/2呈剂量依赖性下调。上述结果表明，大蒜素主要通过诱导晚期凋亡抑制KG-1细胞增殖；凋亡的执行涉及裂解型caspase 3；凋亡的诱导涉及蛋白表达、ERK1/2表达降低、survivin表达以及p-ERK1/2的剂量依赖性降低；mRNA表达涉及Bax增加以及survivin、Bcl-2和ERK1/2下调。
