Meng Xian-Fang, Zou Xiao-Jing, Peng Bin, Shi Jing, Guan Xin-Min, Zhang Chun
Department of Neurobiology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Acta Pharmacol Sin. 2006 Jun;27(6):659-64. doi: 10.1111/j.1745-7254.2006.00324.x.
To observe the effects of tanshinone IIA (Tan IIA) on the neurotoxicity induced by ethanol in PC12 cells and to explore its protective role.
PC12 cell survival was measured by MTT assay. The formation of reactive oxygen species (ROS) and lactate dehydrogenase (LDH) release were detected by 2',7'-dichlorofluorescin (DCF) fluorescence and calorimetric method, respectively. The percentage of cell apoptosis was monitored by flow cytometry. The expression of p53 was detected by immuno-fluorescence and flow cytometry.
Ethanol significantly impaired the survival of PC12 cells as demonstrated by MTT assay. Ethanol also induced significant ROS formation and increased LDH release. Pre-incubation with Tan IIA in the culture medium significantly reversed these changes. Ethanol caused cell apoptosis and the upregulation of p53 protein. The anti-apoptosis effects of Tan IIA on ethanol-induced toxicity were accompanied by the downregulation of pro-apoptotic p53 protein expression.
Tan IIA can protect neurons from apoptosis and might serve as a potential therapeutic drug for neurological disorders induced by ethanol.
观察丹参酮IIA（Tan IIA）对乙醇诱导的PC12细胞神经毒性的影响，并探讨其保护作用。
采用MTT法检测PC12细胞存活率。分别用2',7'-二氯荧光素（DCF）荧光法和比色法检测活性氧（ROS）的生成及乳酸脱氢酶（LDH）的释放。通过流式细胞术监测细胞凋亡率。采用免疫荧光和流式细胞术检测p53的表达。
MTT法显示乙醇显著损害PC12细胞的存活率。乙醇还诱导大量ROS生成并增加LDH释放。在培养基中预孵育Tan IIA可显著逆转这些变化。乙醇导致细胞凋亡并使p53蛋白上调。Tan IIA对乙醇诱导毒性的抗凋亡作用伴随着促凋亡p53蛋白表达的下调。
Tan IIA可保护神经元免于凋亡，可能作为乙醇诱导的神经疾病的潜在治疗药物。
