消栓肠溶胶囊对缺氧缺糖再灌注损伤PC12细胞的保护作用

Protective Effect of Xiaoshuan Enteric Capsule( 消栓肠溶胶囊) on Adrenal Pheochromocytoma Cells( PC12 ) Injury Induced by Hypoxia and Glucose Deprivation

目的:观察消栓肠溶胶囊对缺氧缺糖再灌注损伤肾上腺嗜铬细胞瘤细胞(PC12细胞)的作用。方法:以PC12细胞缺氧缺糖再灌注损伤模拟脑缺血再灌注损伤,研究消栓肠溶胶囊在0. 01～10. 00 mg/L浓度范围内对其的作用,采用噻唑蓝法(MTT法)检测PC12细胞的存活率;乳酸脱氢酶(LDH)法检测PC12细胞损伤程度;酶联免疫(ELISA)法检测基质金属蛋白酶-9(MMP-9)和凋亡相关因子P53、B细胞淋巴瘤/白血病-2(B-cell lymphoma/leukemia-2,BCL-2)、BCL-2关联X蛋白(bcl2-associated X protein,Bax)和半胱氨酸蛋白水解酶(Caspase-3)的水平;逆转录-聚合酶链反应(RT-PCR)法检测p53 mRNA、Bax mRNA和caspase-3 mRNA的表达;分光光度法检测超氧化歧化酶(SOD)和丙二醛(MDA)的水平,以及采用荧光酶标仪检测活性氧(ROS)的表达。结果:与模型组比较,消栓肠溶胶囊各剂量组干预后,缺氧缺糖再灌注诱导的PC12细胞的存活率显著增加(P <0. 05),细胞外LDH比例显著减少(P <0. 05),MMP-9、P53、Bax、Caspase-3、MDA和ROS表达都显著减少(P <0. 05),而BCL-2表达和SOD活性都显著增加(P <0. 05)。结论:消栓肠溶胶囊对缺氧缺糖再灌注诱导的PC12细胞损伤具有明显保护作用,能减少细胞凋亡相关因子表达,其机制可能与抑制MMP-9表达和减轻氧化应激损伤有关。

Objective:To observe the effects of Xiaoshuan Entric Capsule on adrenal pheochromocytoma cells (PC12cells)injury induced by hypoxia and glucose deprivation.Methods:The effects of Xiaoshuan Entric Capsule in the concentration range of 0.01~10.00mg /L on cerebral ischemia -reperfusion injury induced by hypoxia /hypoglycemia injury in PC12cells,the survival rate of PC12cells was detected by MTT assay,the in- jury degree of PC12 cells was detected by lactate dehydrogenase (LDH),the expressions of matrix metalloprotei- nase -9(MMP -9)and apoptosis factors (P53,BCL -2,Bax and Caspase -3)were detected by ELISA,the expressions of p53,Bax ,caspase-3mRNA were detected by RT-PCR,the superoxide dismutase (SOD)and malondialdehyde (MDA),and the expression of reactive oxygen species (ROS)were deteeted by microplate reader.Results:Compared with the model group,in the different doses groups of Xiaoshuan Enterie Capsule, the survival rate of PC12cells induced by hypoxia /glucose deprivation was significantly increased (P <0.05), the ratio of extracellular LDH was significantly decreased (P <0.05),while the expressions of MMP-9,P53,Bax,Caspase-3,MDA and ROS were significantly decreased (P<0.05),while the expressions of BCL -2 and SOD were significantly increased (P <0.05).Con- clusion:Xiaoshuan Enteric Capsule has obvious protective effect on PC12 cell injury induced by hypoxia and glucose deprivation,which can reduce the expression of apoptosis-related factors,which,may be related to the inhibition of MMP-9 expression and the reduction of oxida-tive stress.