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大蒜素对KG-1细胞增殖和凋亡的影响及其分子机制

[Effect of allicin on proliferation and apoptosis of KG-1 cells and its molecular mechanism].

作者信息

Lou Zheng, Wei Qing-Qing, Wang Da-Wei, Gu Huan-Peng

机构信息

The 2nd Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou 310053, China.

School of Stomatology, Zhejiang Chinese Medical University, Hangzhou 310053, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2018 Jun;43(12):2612-2617. doi: 10.19540/j.cnki.cjcmm.20180320.002.

Abstract

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.

摘要

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。

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