次野鸢尾黄素抗缺血性脑卒中活性研究

On the Anti-Ischemic Stroke Effect of Subnormal Irisflorentin

探讨次野鸢尾黄素对Na_(2)S_(2)O_(4)诱导的糖氧剥夺体外脑卒中模型损伤的保护作用机制。将PC12细胞分为空白组(不给药,更换正常培养液培养24 h)、模型组(加入10μg/mL Na_(2)S_(2)O_(4)缺氧诱导2 h后,更换正常培养液培养24 h)、阳性对照组(加入10μg/mL Na_(2)S_(2)O_(4)缺氧诱导2 h后,更换含10μg/mL依达拉奉培养24 h)及药物保护组(加入10μg/mL Na_(2)S_(2)O_(4)缺氧诱导2 h后,更换含3.13、6.25、12.5μg/mL次野鸢尾黄素培养24 h),观察细胞形态、检测细胞存活率,LDH释放量,细胞内Caspase-3活性、Ca 2+水平、细胞凋亡情况,免疫印迹法检测凋亡蛋白Caspase-3的表达水平。结果显示,射干中次野鸢尾黄素能显著降低氧糖剥夺/复氧对PC12细胞造成的损伤程度,提升细胞存活率,降低细胞内LDH释放量、Ca 2+含量、Caspase-3活性,降低Caspase-3蛋白表达量。结果表明,次野鸢尾黄素对氧糖剥夺/复氧损伤的PC12细胞有显著的保护作用(P<0.01),其机理可能与抑制脑内神经细胞的凋亡相关。

To investigate the protective mechanism of irisflorentin on Na_(2)S_(2)O_(4)-induced injury in an in vitro stroke model,the PC12 cells were divided into blank group(no drug administration,replaced with normal culture medium for 24 h),model group(cultured in normal culture medium for 24 h after adding 10μg/mL Na_(2)S_(2)O_(4) for 2 h),positive control group(cultured in 10μg/mL edaravone for 24 h after adding 10μg/mL Na_(2)S_(2)O_(4) for 2 h),and drug-protected group(after the induction of hypoxia by adding 10μg/mL Na_(2)S_(2)O_(4) for 2 h,and then replacing the incubation with 3.13,6.25 and 12.5μg/mL subnormal irisflorentin for 24 h).And the morphology of the cells was observed,cell viability was examined;and the amount of LDH released,intracellular Caspase-3 enzyme activity,Ca 2+level,and apoptosis were detected.The expression level of apoptotic protein Caspase-3 was detected by immunoblotting.The results showed that secondary wild irisflorentin in Belamcanda chinensis significantly reduced the degree of damage caused by oxygen glucose deprivation/reoxygenation to PC12 cells,enhanced cell survival,decreased intracellular LDH release,Ca 2+level,as well as Caspase-3 enzyme activity,and decreased the expression level of Caspase-3 protein.The results showed that Irisflorentin had a significant protective effect(P<0.01)on oxygen-glucose deprivation/reoxygenation-injured PC12 cells,and its mechanism might be related to the inhibition of apoptosis of neuronal cells in the brain.