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目的:观察中药心脉通颗粒含药血清对原代培养大鼠心肌细胞H2O2损伤的抗氧化作用。方法:原代培养的大鼠心肌细胞分为3组,正常对照组用正常大鼠血清培养,模型组用正常大鼠血清培养后,H2O2造损伤模型。心脉通组用心脉通含药血清干预12 h后,再加H2O2造损伤。各组分别在H2O2作用6h后检测实验中各指标:心肌细胞形态学的改变;MTT法检测细胞活性的改变;细胞免疫组织化学法检测蛋白激酶B(PKB/Akt)的表达;RT-PCR检测蛋白激酶B(PKB/Akt)和内皮细胞型一氧化氮合酶(e NOS)的基因表达变化。结果:各组心肌细胞在H2O2作用下活力下降,与正常对照组比较,模型组细胞活力显著下降,差异有统计学意义(P<0.05),表明H2O2损伤模型建立成功;心脉通组细胞活力高于模型组,差异有统计学意义(P<0.05)。在正常状态下PKB的转录水平较低,模型组有所升高,与正常对照组比较,差异有统计学意义(P<0.05)。且心脉通组的转录水平更高,与正常对照组和模型组比较,差异均有统计学意义(P<0.05)。正常情况下e NOS有表达,且表达水平是最高的。模型组中表达降低,与正常对照组比较,差异有统计学意义(P<0.05)。心脉通组转录水平较模型组水平增高(P<0.05)。结论:中药复方心脉通颗粒能够增强心肌细胞抗氧化损伤能力,保持细胞的形态和功能,抑制细胞调亡,其机制可能是通过PKB/AKT-e NOS信号传导途径。
OBJECTIVE: To observe the anti-oxidative effect of Xinmaitong granule-containing serum on H2O2 injury in primary cultured rat cardiac myocytes. Methods: Primary cultured rat cardiomyocytes were divided into three groups. The normal control group was cultured in normal rat serum. The model group was cultured in normal rat serum and then induced by H2O2. Xinthi Tongxinluo intravenous drug-containing serum intervention 12 h, plus H2O2 injury. The changes of cell morphology were detected by MTT assay, the expression of PKB / Akt was detected by immunohistochemistry and the expression of PKB / Akt was detected by RT-PCR Protein kinase B (PKB / Akt) and endothelial cell nitric oxide synthase (eNOS) gene expression changes. Results: Compared with normal control group, the viability of cardiomyocytes in each group decreased under the action of H2O2. Compared with normal control group, the viability of model group decreased significantly (P <0.05), indicating that H2O2 injury model was established successfully. Higher than the model group, the difference was statistically significant (P <0.05). Under normal conditions, the transcription level of PKB was lower in model group than in normal control group, with significant difference (P <0.05). The Xinthi tong group had a higher level of transcription, compared with the normal control group and the model group, the differences were statistically significant (P <0.05). Under normal circumstances eNOS expression, and the expression level is the highest. The expression in model group decreased, compared with the normal control group, the difference was statistically significant (P <0.05). Xinmaitong group had a higher level of transcription than the model group (P <0.05). Conclusion: The Chinese medicine compound Xinmaitong Granule can enhance the ability of cardiomyocytes against oxidative damage, maintain the morphology and function of cells, and inhibit the apoptosis of cells. The mechanism may be through the PKB / AKT-e NOS signal transduction pathway.