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目的探讨当归红芪超滤物对H2O2致乳鼠心肌细胞氧化损伤的保护作用及其可能机制。方法用高浓度的H2O2(400μmol/L)在Wistar乳鼠原代培养的心肌细胞上建立氧化损伤模型,用不同质量浓度的当归红芪超滤物(3.75、7.5、15mg/mL)进行干预。在倒置显微镜下观察各实验组心肌细胞的搏动频率,用MTT法检测心肌细胞的存活率,用试剂盒检测细胞培养上清液中乳酸脱氢酶(LDH)、肌酸激酶(CK)的活性及细胞内超氧化物歧化酶(SOD)的活性、丙二醛(MDA)、髓过氧化物酶(MPO)的量。RT-PCR技术检测caspase-3、hsp70基因在心肌细胞中的表达情况。结果当归红芪超滤物显著提高氧化损伤心肌细胞的细胞活力;与损伤组比较,当归红芪超滤物各干预组心肌细胞LDH、CK、MPO、MDA量显著降低(P<0.01),SOD活性显著提高(P<0.01),caspase-3基因表达显著降低(P<0.05),hsp70基因表达显著提高(P<0.05),差异具有统计学意义;并且当归红芪超滤物的抗氧化损伤作用呈剂量依赖性。结论当归红芪超滤物具有良好的抗氧化损伤作用,其机制可能与清除自由基、上调hsp70表达、抑制caspase-3活性有关。
Objective To investigate the protective effect of angelica Hedysari ultrafiltration on oxidative damage of neonatal rat cardiomyocytes induced by H2O2 and its possible mechanism. Methods Oxidative damage was induced by high concentration of H2O2 (400μmol / L) on primary cultured cardiomyocytes of Wistar suckling mice. The effects of different concentrations of Radix Angelicae Sinensis ultrafiltration (3.75,7.5,15 mg / mL) were also investigated. The beating frequency of cardiomyocytes in each experimental group was observed under an inverted microscope. The survival rate of cardiomyocytes was measured by MTT assay. The activity of lactate dehydrogenase (LDH) and creatine kinase (CK) in the cell culture supernatant was detected by kit And intracellular superoxide dismutase (SOD) activity, the amount of malondialdehyde (MDA), myeloperoxidase (MPO). RT-PCR was used to detect the expression of caspase-3 and hsp70 in cardiomyocytes. Results Angelica sinensis ultrafiltration significantly increased the cell viability of oxidative injured cardiomyocytes. Compared with the injury group, the LDH, CK, MPO and MDA of the myocardial cells decreased significantly (P <0.01) (P <0.01), the expression of caspase-3 gene was significantly decreased (P <0.05), the expression of hsp70 gene was significantly increased (P <0.05), and the difference was statistically significant; and the anti-oxidative damage The effect was dose-dependent. Conclusion Angelica sinensis ultrafiltration has a good anti-oxidative damage effect, its mechanism may be related to scavenging free radicals, up-regulating hsp70 expression and inhibiting caspase-3 activity.