苦参碱对体外诱导人结肠HT29细胞凋亡作用的实验研究

常城; 张海; 王珍; 刘少平

doi:10.3969/j.issn.1671-038X.2013.12.004

苦参碱对体外诱导人结肠HT29细胞凋亡作用的实验研究

- 1. 湖北理工学院附属黄石中心医院消化内科, 湖北黄石 435000;
- 2. 湖北理工学院附属黄石中心医院放射科, 湖北黄石 435000;
- 3. 湖北理工学院附属黄石中心医院湖北省肾脏疾病发生与干预重点实验室, 湖北黄石 435003
基金项目:
湖北省自然科学基金资助项目(No:2010CDB07401)

详细信息

作者简介:
常城,男,医学博士,主治医师,从事消化系统肿瘤的防治研究;

通讯作者:张海 E-mail:hubeitcm@sohu.com

通讯作者: 张海E-mail:hubeitcm@sohu.com

中图分类号: R284.1;R329.2

收稿日期: 2013-08-11

Matrine induces apoptosis of human colon cancer HT29 cells through mitochondrial pathway

- 1. Department of Gastroenterology, Huangshi Central Hospital of Hubei;Polytechnic University, Huangshi 435000, China;
- 2. Department of Radiology, Huangshi Central Hospital of Hubei Polytechnic University, Huangshi 435000, China;
- 3. Hubei Key Laboratory of Kidney Disease Pathogenesis and Intervention, Huangshi 435003, China

More Information

Corresponding author: ZHANG Hai, E-mail: hubeitcm@sohu.com

Received Date: 11 August 2013

摘要

摘要: [目的]研究苦参碱(matrine,MA)对体外诱导人结肠癌HT29细胞的凋亡作用及其机制。[方法] 不同浓度MA(0~32 mg/ml)作用HT29细胞24 h后,流式细胞仪检测细胞凋亡,线粒体跨膜电位检测试剂盒(JC-1)检测线粒体膜电位(Δφm)变化,分光光度法检测caspase-3,-9酶活性,Western blot法检测线粒体凋亡途径相关蛋白Bax、Bcl-2表达。[结果]MA显著诱导HT29细胞凋亡;16 mg/ml MA作用HT29细胞24 h后,caspase-9,-3酶活性明显升高;Western blot结果显示:MA处理组促凋亡蛋白Bax表达明显升高,抗凋亡蛋白Bcl-2表达明显降低。[结论] MA具有促进结肠癌细胞凋亡的作用,且具有剂量依赖性,其机制与线粒体凋亡途径有关。
- 苦参碱 /
- 细胞凋亡 /
- 实验研究
Abstract: [Objective][WT5BZ] To investigate the apoptosis-inducing effect of Matrine (MA) on HT29 cells and the underlying mechanism.[WT5HZ][Methods][WT5BZ] HT29 cells were treated with MA of 0–16mg/ml concentration.The apoptosis was determined by annexin V-FITC/propidium iodide assay.The mitochondrial membrane potential(Δφm) was evaluated by flow cytometry with JC-1 as probe.Protein levels were determined by western blot analysis and the role of caspase activities were assayed using caspase apoptosis detection kit.[WT5HZ][Results][WT5BZ] After treatment with MA,a decrease in the viability of HT29 cells and increase in apoptosis were observed.MA-induced apoptosis was accompanied by loss of mitochondrial membrane potential.Meanwhile,the anti-apoptotic protein Bcl-2 level was down-regulated,whereas,the pro-apoptotic protein Bax level up-regulated.MA increased the activity of caspase-9 and caspase-3.[WT5HZ][Conclusion][WT5BZ] These results indicate that MA can induce the apoptosis of HT29 cell lines in a dose-dependent manner,which is probably mediated by mitochondrial apoptosis pathway.
[WT5HZ] Key words:[WT5BZ]Matrine; apoptosis; experimental research
[HK][HT]

[FL(K2]
- [Objective] To investigate the apoptosis-inducing effect of Matrine (MA) on HT29 cells and the underlying mechanism.[Methods] HT29 cells were treated with MA of 0-16mg/ml concentration.The apoptosis was determined by annexin V-FITC/propidium iodide assay.The mitochondrial membrane potential(Δφm) was evaluated by flow cytometry with JC-1 as probe.Protein levels were determined by western blot analysis and the role of caspase activities were assayed using caspase apoptosis detection kit.[Results] After treatment with MA,a decrease in the viability of HT29 cells and increase in apoptosis were observed.MA-induced apoptosis was accompanied by loss of mitochondrial membrane potential.Meanwhile,the anti-apoptotic protein Bcl-2 level was down-regulated,whereas,the pro-apoptotic protein Bax level up-regulated.MA increased the activity of caspase-9 and caspase-3.[Conclusion] These results indicate that MA can induce the apoptosis of HT29 cell lines in a dose-dependent manner,which is probably mediated by mitochondrial apoptosis pathway. /

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