Thakur Rohit Singh, Devaraj Ezhilarasan
Department of Pharmacology, Malla Reddy Institute of Medical Sciences, Hyderabad, India.
Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Chennai, Tamil Nadu, India.
Environ Toxicol. 2020 Nov;35(11):1225-1233. doi: 10.1002/tox.22987. Epub 2020 Jul 22.
Hepatocellular carcinoma is the second leading cause of cancer-related mortality worldwide. Lagerstroemia speciosa Pers. (Lythraceae) commonly known as Banaba has been used in different forms in traditional medicinal systems for treating various diseases which include diabetes and obesity. In this study, we investigated the cytotoxic potential of ethanolic Banaba leaf extract (EBLE) in HepG2 cells. The phytochemical analysis of EBLE was performed by HPTLC. HepG2 cells were treated with EBLE at 25, 50, 100, and 150 μg/mL concentrations, and cytotoxicity was evaluated by MTT assay. Oxidative stress was assessed by the evaluation of lipid peroxidation, superoxide dismutase, and reduced glutathione. Apoptosis-related morphology was investigated by acridine orange and ethidium bromide (AO/EB) dual staining. Mitochondrial membrane potential (ΔΨm) was evaluated by JC-1 staining. Apoptosis-related marker genes were evaluated by qPCR. HPTLC analysis confirmed the presence of corosolic acid (12.87 μg/mg), berberine (3.19 μg/mg), and gallic acid (2.94 μg/mg) in EBLE. EBLE treatments caused significant and concentration-dependent cytotoxicity and oxidative stress in HepG2 cells. Dual staining with AO/EB confirmed membrane distortion and nuclear chromatin condensation upon EBLE treatments. JC-I staining revealed the loss of ΔΨm. Furthermore, at a molecular level, EBLE treatments interfere with Bax/Bcl-2 homeostasis and induced the pro-apoptotic marker genes such as cytochrome c, Apaf-1, and caspases 9 and 3. EBLE treatments caused cytotoxicity in HepG2 cells, and this could be due to the induction of oxidative stress and apoptosis via the intrinsic or mitochondrial pathway.
肝细胞癌是全球癌症相关死亡的第二大主要原因。紫薇（千屈菜科）俗称巴拿巴，在传统医学体系中以不同形式用于治疗包括糖尿病和肥胖症在内的各种疾病。在本研究中，我们研究了巴拿巴叶乙醇提取物（EBLE）对HepG2细胞的细胞毒性潜力。通过高效薄层层析法对EBLE进行了植物化学分析。用浓度为25、50、100和150μg/mL的EBLE处理HepG2细胞，并通过MTT法评估细胞毒性。通过评估脂质过氧化、超氧化物歧化酶和还原型谷胱甘肽来评估氧化应激。通过吖啶橙和溴化乙锭（AO/EB）双重染色研究凋亡相关形态。通过JC-1染色评估线粒体膜电位（ΔΨm）。通过定量聚合酶链反应评估凋亡相关标记基因。高效薄层层析分析证实EBLE中存在熊果酸（12.87μg/mg）、小檗碱（3.19μg/mg）和没食子酸（2.94μg/mg）。EBLE处理在HepG2细胞中引起显著的浓度依赖性细胞毒性和氧化应激。AO/EB双重染色证实EBLE处理后细胞膜变形和核染色质浓缩。JC-I染色显示ΔΨm丧失。此外，在分子水平上，EBLE处理干扰Bax/Bcl-2稳态并诱导促凋亡标记基因，如细胞色素c、凋亡蛋白酶激活因子-1以及半胱天冬酶9和3。EBLE处理在HepG2细胞中引起细胞毒性，这可能是由于通过内在或线粒体途径诱导氧化应激和凋亡所致。
