【作者】 韩凤昭；

【导师】 邹莉波；

【作者基本信息】 沈阳药科大学 ， 药理学， 2008， 硕士

【摘要】 目的:对以红景天苷为先导化合物合成的一系列化合物进行抗缺血缺氧活性筛选,期望从大量的化合物中筛选出具有抗缺血缺氧活性的物质并为新药的开发提供科学的实验依据。方法:（1）不同浓度药物预处理细胞后制备原代心肌细胞缺氧—复氧模型和谷氨酸诱导的SH-SY5Y细胞损伤模型,全自动生化分析仪检测乳酸脱氢酶（LDH）释放抑制率的变化。（2）选出其中活性较好、合成方法简单的S01为代表化合物,初步研究其药效及抗缺血缺氧作用机制。以不同浓度S01药物(2,10 and 50μg·ml^-1)预处理心肌细胞或者神经细胞后,建立心肌细胞的缺氧—复氧损伤或者谷氨酸诱导的SH-SY5Y细胞损伤模型,检测心肌细胞上清液中的乳酸脱氢酶（LDH）、天冬氨酸转氨酶（AST）、肌酸激酶（CK）的含量和细胞凋亡程度及SH-SY5Y细胞上清中乳酸脱氢酶（LDH）含量和细胞存活率,同时检测细胞内Ca²⁺浓度、活性氧（ROS）水平、ATP水平,及细胞上清中NO水平、超氧化物歧化酶（SOD）活性和丙二醛含量及S01对正常小鼠游泳耐疲劳能力的影响和常压耐缺氧能力的影响。结果（1）利用细胞模型的筛选,发现有4种化合物的体外抗缺血缺氧活性优于先导化合物。（2）预先给予S01能够降低损伤细胞中乳酸脱氢酶、天冬氨酸转氨酶（AST）、肌酸激酶（CK）的含量和细胞凋亡程度及SH-SY5Y细胞上清中乳酸脱氢酶（LDH）含量,升高细胞存活率,改善细胞功能。另外结果表明,S01能够升高SOD活性,且可减小胞内ATP水平降低程度,且能降低损伤细胞中ROS升高水平、MDA含量、NO水平、细胞内钙超载和细胞凋亡,从而保护细胞,同时S01能够显著延长小鼠的存活时间,增强小鼠耐常压缺氧的能力。结论:这些化合物通过多种机制来对缺血缺氧的细胞进行保护作用,可能对脑缺血或者心肌缺血具有治疗作用。更多还原

【Abstract】 AIM（1）To find compounds possessing anti-ischemia and anti-anoxia activity from compounds derived from Salidroside and provides the scientific experimental data for newdrug -research.METHODS（1）SH-SY5Y cells and cultured neonatal rat myocardial cells were pretreated with each compound(40,200 and 1000 ug·ml^-1,respectively)and then induced damage by glutamate or hypoxia/reoxygenation,the inhibition rate of Lactate dehydrogenase（LDH） release was detected by colorimetry.（2）S01 which has better acticity and simpler synthesis method was selected as the representative to research the drug action and anti-ischemia/anti-anoxia mechanism.SH-SY5Y cells and cultured neonatal rat myocardial cells were pretreated with S01(2,10 and 50 ug·ml^-1,respectively)and then induced damage by glutamate or hypoxia/reoxygenation,then the content of LDH,AST,CK enzymes and the cell apoptosis degree in cultured neonatal rat myocardia cells,the content of LDH enzyme and cell viability in SH-SY5Y cells,cell the free Ca²⁺concentration([Ca²⁺]_i),intracellular reactive oxygen species（ROS）level,adenosine triphosphate（ATP）level,nitric oxide（NO）level, oxide dismutase（SOD）activity and malondialdehyde（MDA）level were detected.The anti-fatigue and anti-hypoxia ability of mice were also detected.RESULTS（1）4 compounds were found to have better anti-ischemia and anti-anoxia activity than lead compounds.（2）Compared with model groups,in cultured neonatal rat myocardial cells,S01(2,10 and 50 ug·ml^-1)can reduce LDH,AST,CK release induced by hypoxia/reoxygenation,in SH-SY5Y cells,S01 can reduce LDH release and levated the cell survival rate induced by glutamate.Moreover,results indicate that S01 can increase SOD activity and ATP level,and inhibited the increase of MDA,ROS,Ca²⁺overload,apoptosis and NO levels induced by glutamate or hypoxia/reoxygenation.Furthermore,S01 can increase the anti-fatigue and anti-hypoxia ability of mice. CONCLUSION The compounds can protect cells from the damage induced by glutamate and oxygen-glucose deprivation through some mechanism,which indicates that they may have the potential to treat cerebral or myocardium ischemia.更多还原