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野罂粟提取物诱导人结肠癌细胞凋亡的作用

Apoptotic-Induced Effects of Willd. Extract in Human Colon Cancer Cells.

机构信息

Dipartimento di Scienze della Vita, Università di Siena, Via Aldo Moro 2, 53100 Siena, Italy.

Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum, Università di Bologna, Via Belmeloro 6, 40126 Bologna, Italy.

出版信息

Int J Mol Sci. 2020 Mar 19;21(6):2102. doi: 10.3390/ijms21062102.

Abstract

The research for innovative treatments against colon adenocarcinomas is still a great challenge. Willd. heartwood extract (AC) has health-promoting qualities, especially at the gastrointestinal level. This study characterized AC for its catechins content and investigated the apoptosis-enhancing effect in human colorectal adenocarcinoma HT-29 cells, along with its ability to spare healthy tissue. MTT assay was used to describe the time course, concentration dependence and reversibility of AC-mediated cytotoxicity. Cell cycle analysis and AV-PI and DAPI-staining were performed to evaluate apoptosis, together with ROS formation, mitochondrial membrane potential (MMP) changes and caspase activities. Rat ileum and colon rings were tested for their viability and functionality to explore AC effects on healthy tissue. Quantitative analysis highlighted that AC was rich in (±)-catechin (31.5 ± 0.82 mg/g) and (-)-epicatechin (12.5 ± 0.42 mg/g). AC irreversibly decreased cell viability in a concentration-dependent, but not time-dependent fashion. Cytotoxicity was accompanied by increases in apoptotic cells and ROS, a reduction in MMP and increases in caspase-9 and 3 activities. AC did not affect rat ileum and colon rings' viability and functionality, suggesting a safe profile toward healthy tissue. The present findings outline the potential of AC for colon cancer treatment.

摘要

针对结肠腺癌的创新治疗方法的研究仍然是一个巨大的挑战。Willd. 心材提取物 (AC) 具有促进健康的特性,特别是在胃肠道水平。本研究对 AC 的儿茶素含量进行了表征,并研究了其在人结直肠腺癌细胞 HT-29 中的促进细胞凋亡作用,以及对健康组织的保护作用。MTT 测定法用于描述 AC 介导的细胞毒性的时间过程、浓度依赖性和可逆转性。通过 AV-PI 和 DAPI 染色进行细胞周期分析,以评估细胞凋亡,同时评估 ROS 形成、线粒体膜电位 (MMP) 变化和半胱天冬酶活性。大鼠回肠和结肠环用于评估其活力和功能,以探索 AC 对健康组织的影响。定量分析突出表明,AC 富含(±)-儿茶素(31.5 ± 0.82mg/g)和(-)-表儿茶素(12.5 ± 0.42mg/g)。AC 以浓度依赖性而非时间依赖性的方式不可逆地降低细胞活力。细胞毒性伴随着凋亡细胞和 ROS 的增加、MMP 的减少以及 caspase-9 和 3 活性的增加。AC 不影响大鼠回肠和结肠环的活力和功能,这表明对健康组织具有安全的特性。本研究结果概述了 AC 治疗结肠癌的潜力。

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