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灵芝诱导 NB4 人白血病细胞凋亡:涉及 Akt 和 Erk。

Ganoderma lucidum induced apoptosis in NB4 human leukemia cells: involvement of Akt and Erk.

机构信息

Departamento de Bioquímica y Biología Molecular, Campus Universitario, Universidad de Alcalá, 28871 Alcalá de Henares, Madrid, Spain.

出版信息

J Ethnopharmacol. 2010 Mar 2;128(1):71-8. doi: 10.1016/j.jep.2009.12.027. Epub 2009 Dec 29.

Abstract

AIM OF THE STUDY

The final goal of this work was to study the toxic and apoptosis effects induced by fractions from Ganoderma lucidum [Ganoderma lucidum (Curtis) P. Karst.; Ganodermataceae Donk] on NB4 human leukemia cells.

MATERIALS AND METHODS

Two aqueous extracts and a methanol-extracted column-chromatography semipurified fraction were obtained from Ganoderma lucidum fruiting body. Flow cytometry analyses were used to measure cell viability, cell cycle and DNA fragmentation and to quantify apoptosis. Western-blot analyses were used to quantify changes in apoptosis proteins and intracellular kinases.

RESULTS

Aqueous extracts slightly reduce cell viability and induce DNA fragmentation in NB4 cells. Methanol-extracted semipurified fraction at dilutions down to 15% or 40% of the initial fraction concentration reduced significantly the viability of these leukemia cells (treated for 19h) with induction of DNA fragmentation and induction of apoptosis. Overmore, the dilution down to 15% of the initial E3 concentration induced a reduction of p53 levels, of the Bcl2/Bax relationship as well as reduced levels of both unphosphorylated and phosphorylated Akt (Protein kinase Akt, protein kinase B) and Erk (Erk1 and 2).

CONCLUSIONS

Induction of apoptosis and alterations in signal transduction kinases (Akt and Erk) are produced by active fractions from Ganoderma lucidum on human leukemia cells. These data could be of important relevance from the viewpoint of antitumor actions of compounds from Ganoderma lucidum. Eventual therapy applications in leukemia cells might be developed.

摘要

研究目的

本研究的最终目的是研究灵芝[灵芝(Curtis)P. Karst。;灵芝科]各部位提取物对 NB4 人白血病细胞的毒性和凋亡作用。

材料与方法

从灵芝子实体中获得两种水提物和一种甲醇提取的柱层析半纯化馏分。采用流式细胞术分析测定细胞活力、细胞周期和 DNA 片段化,并定量细胞凋亡。Western-blot 分析用于定量凋亡蛋白和细胞内激酶的变化。

结果

水提物轻微降低 NB4 细胞的活力并诱导 DNA 片段化。甲醇提取的半纯化馏分在初始馏分浓度的 15%或 40%稀释度下,显著降低这些白血病细胞(处理 19 小时)的活力,导致 DNA 片段化和凋亡诱导。此外,E3 初始浓度的 15%稀释度可降低 p53 水平、Bcl2/Bax 比值以及未磷酸化和磷酸化 Akt(蛋白激酶 Akt,蛋白激酶 B)和 Erk(Erk1 和 2)的水平。

结论

灵芝活性部位诱导人白血病细胞凋亡和信号转导激酶(Akt 和 Erk)改变。这些数据从灵芝化合物的抗肿瘤作用角度来看可能具有重要意义。最终可能会在白血病细胞中开发出治疗应用。

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