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解毒消癥饮通过线粒体介导的途径诱导细胞凋亡来抑制肝癌细胞增殖。

Jiedu Xiaozheng Yin decoction inhibits hepatoma cell proliferation by inducing apoptosis via the mitochondrial-mediated pathway.

作者信息

Cao Zhiyun, Chen Xuzheng, Lin Wei, Zhao Jinyan, Zheng Liangpu, Ye Hongzhi, Liao Lianming, Du Jian

机构信息

Fujian Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350108, P.R. China.

出版信息

Mol Med Rep. 2015 Aug;12(2):2800-6. doi: 10.3892/mmr.2015.3696. Epub 2015 Apr 28.

DOI:10.3892/mmr.2015.3696
PMID:25936398
Abstract

Jiedu Xiaozheng Yin decoction (JXY) is a type of Chinese traditional medicine, which has been used to treat various types of cancer. The present study explored the mechanisms underlying the anticancer activity of JXY. The effects of ethyl acetate extraction of JXY (EE-JXY) were evaluated on the HepG2 human hepatoma cell line in vitro and in vivo. Following treatment of the HepG2 cells with EE-JXY for 24 h, cell viability, apoptosis, mitochondrial membrane potential, caspase enzyme activity and the expression levels of apoptotic-associated proteins (Bcl-2 and Bax) were detected by MTT, flow cytometry, ELISA and western blotting respectively. In addition, HepG2 cells were subcutaneously transplanted into BALB/c nude mice, and the tumor bearing mice were treated with either EE-JXY (0.06 g/kg) or normal saline for 21 days. Tumor volume and weight were measured and recorded. The apoptotic index, and the expression levels of Bax and cytochrome c were determined with immunohistochemical staining. Treatment with EE-JXY inhibited the proliferation of HepG2 cells, and reduced cell viability in a dose-and time-dependent manner. Furthermore, EE-JXY induced HepG2 cell apoptosis, as demonstrated by a loss of plasma membrane asymmetry and externalization of phosphatidylserine, collapse of mitochondrial membrane potential, activation of caspase-9 and caspase-3, and an increased ratio of pro-apoptotic Bax to anti-apoptotic Bcl-2. Furthermore, EE-JXY inhibited tumor growth and increased the apoptotic index of tumors in tumor-bearing mice. In conclusion, the results of the present study suggest that JXY inhibits HepG2 cell proliferation through mitochondrion-mediated apoptosis, which may partially explain its anticancer activity.

摘要

解毒消症饮(JXY)是一种中药,已被用于治疗各种类型的癌症。本研究探讨了JXY抗癌活性的潜在机制。对JXY的乙酸乙酯提取物(EE-JXY)在体外和体内对HepG2人肝癌细胞系的作用进行了评估。用EE-JXY处理HepG2细胞24小时后,分别通过MTT、流式细胞术、ELISA和蛋白质印迹法检测细胞活力、凋亡、线粒体膜电位、半胱天冬酶活性以及凋亡相关蛋白(Bcl-2和Bax)的表达水平。此外,将HepG2细胞皮下移植到BALB/c裸鼠体内,对荷瘤小鼠用EE-JXY(0.06 g/kg)或生理盐水处理21天。测量并记录肿瘤体积和重量。用免疫组织化学染色法测定凋亡指数以及Bax和细胞色素c的表达水平。EE-JXY处理抑制了HepG2细胞的增殖,并以剂量和时间依赖性方式降低细胞活力。此外,EE-JXY诱导HepG2细胞凋亡,表现为质膜不对称性丧失和磷脂酰丝氨酸外化、线粒体膜电位崩溃、半胱天冬酶-9和半胱天冬酶-3激活以及促凋亡Bax与抗凋亡Bcl-2的比例增加。此外,EE-JXY抑制荷瘤小鼠肿瘤生长并增加肿瘤的凋亡指数。总之,本研究结果表明JXY通过线粒体介导的凋亡抑制HepG2细胞增殖,这可能部分解释了其抗癌活性。

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