Department of Neurosurgery, Xi'an Children's Hospital, Xi'an, Shaanxi, China.
Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia.
J Biochem Mol Toxicol. 2021 Sep;35(9):e22857. doi: 10.1002/jbt.22857. Epub 2021 Aug 2.
Gliomas are a type of brain cancer that occurs in the supporting glial cells of the brain. It is highly malignant and accounts for 80% of brain tumors with high mortality and morbidity. Phytomedicines are potent alternatives for allopathic drugs which cause side effects. They have been used from ancient times by traditional Chinese, Ayurveda, and Siddha medicine. Arubtin is a glycoside phytochemical extracted from plants and belongs to the family of Ericaceae. Arbutin possesses various pharmacological properties such as anti-inflammatory, antioxidant, antitumor, and so on. Hence in the present study, we analyzed the anticancer potency of arbutin against rat C6 glioma cells. Rat C6 glioma cells were procured from American Type Culture Collection and the cells were cultured in Roswell Park Memorial Institute-1640 medium. To assess the cytotoxicity effect of the arbutin against C6 glioma cells, an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide test was performed with different doses from 10 to 60 µM. Arbutin effectively induced apoptosis in the cells and the IC dose was obtained at 30 µM. For further studies, we selected the 30 µM IC dose and a higher dose of 40 µM. Reactive oxygen species (ROS) generated were analyzed with DCFDA/H2DCFDA stain and the destruction of mitochondrial membrane permeability which is the initiator of apoptosis was analyzed with a cationic stain Rhodamine 123. Dual staining with acridine orange and ethidium bromide was performed to assess the viable and dead cells. Cell adhesion properties of glioma cells were analyzed with Matrigel assay. The apoptotic, inflammatory, and phosphoinositide 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) signaling molecules were analyzed with quantitative polymerase chain reaction (qPCR) analysis to confirm the anticancer effect of arbutin. Arbutin generated excessive ROS and disrupted the mitochondrial membrane, which induced apoptosis in cells, it also inhibited the cell adhesion property of C6 glioma cells. qPCR analysis clearly indicates arbutin increases the apoptotic genes and decreased the inflammatory and PI3K/mTOR signaling molecules. Overall, our results authentically confirm that arbutin can be a potent alternative for treating glioma.
神经胶质瘤是一种发生在大脑神经胶质细胞中的脑癌。它具有高度恶性，占高死亡率和发病率的脑瘤的 80%。植物药是对抗有副作用的西药的有效替代品。它们自古以来就被中医、阿育吠陀和悉达医学所使用。熊果苷是一种从植物中提取的糖苷类植物化学物质，属于杜鹃花科。熊果苷具有多种药理作用，如抗炎、抗氧化、抗肿瘤等。因此，在本研究中，我们分析了熊果苷对大鼠 C6 神经胶质瘤细胞的抗癌作用。大鼠 C6 神经胶质瘤细胞购自美国典型培养物保藏中心，细胞在罗格斯大学纪念研究所-1640 培养基中培养。为了评估熊果苷对 C6 神经胶质瘤细胞的细胞毒性作用，我们用不同剂量（10-60 μM）进行了 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物试验。熊果苷能有效诱导细胞凋亡，IC 剂量为 30 μM。进一步研究，我们选择了 30 μM 的 IC 剂量和 40 μM 的较高剂量。用 DCFDA/H2DCFDA 染色分析产生的活性氧（ROS），用阳离子染色罗丹明 123 分析凋亡起始的线粒体膜通透性破坏。用吖啶橙和溴化乙锭双重染色评估活细胞和死细胞。用 Matrigel 测定分析神经胶质瘤细胞的黏附特性。用定量聚合酶链反应（qPCR）分析分析凋亡、炎症和磷酸肌醇 3-激酶（PI3K）/哺乳动物雷帕霉素靶蛋白（mTOR）信号分子，以确认熊果苷的抗癌作用。熊果苷产生过多的 ROS 并破坏线粒体膜，从而诱导细胞凋亡，还抑制 C6 神经胶质瘤细胞的黏附特性。qPCR 分析清楚地表明，熊果苷增加了凋亡基因，减少了炎症和 PI3K/mTOR 信号分子。总的来说，我们的结果确凿地证实，熊果苷可以成为治疗神经胶质瘤的有效替代品。
