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目的:观察红花水提物对体外培养大鼠乳鼠心肌细胞H2O2损伤的保护作用及电子自旋共振(ESR)波谱的影响。方法:建立心肌细胞H2O2损伤模型,实验分5组,空白对照组,模型对照组:H2O2(1 mmol.L-1)孵育1 h;红花水提物500,100,20 mg.L-1剂量组:分别给予不同浓度的红花水提物,预处理24 h,然后换为H2O2(1 mmol.L-1)孵育1 h。实验结束后分别收集细胞上清液测定乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、一氧化氮(NO)、一氧化氮合酶(NOS)和黄嘌呤氧化酶(XOD)活性,用自旋捕集技术结合电子自旋共振波谱技术(Electron Spin Res-onance,简称ESR)检测各组细胞悬液中氧自由基信号的强弱。结果:红花水提物各剂量组能够明显提高H2O2损伤后的心肌细胞的搏动频率,使细胞存活率提高,减少心肌细胞LDH的释放,降低培养液中MDA和XOD的含量,同时提高SOD,NO和GSH-Px的活性,使细胞悬液中的ESR氧自由基信号明显减弱。结论:红花水提物对体外培养乳鼠心肌细胞具有保护作用,其机制与其减少活性氧产生、增强氧自由基的清除、增强内源性抗氧化酶的活性有关。
Objective: To observe the protective effect of safflower aqueous extract on the H2O2 injury of neonatal rat cardiomyocytes in vitro and the effect of electron spin resonance (ESR) spectroscopy. Methods: H2O2 injury model of cardiomyocytes was established. The experiment was divided into 5 groups: blank control group, model control group: H2O2 (1 mmol.L-1) for 1 h; safflower water extract 500, 100, 20 mg.L-1 : Different concentrations of safflower aqueous extract were pretreated for 24 h, then changed to H2O2 (1 mmol.L-1) for 1 h. After the experiment, the cell supernatants were collected to determine the content of LDH, MDA, GSH-Px, Nitrogen (NO), nitric oxide synthase (NOS) and xanthine oxidase (XOD) activity were detected by using spin trapping technique combined with Electron Spin Resonance (ESR) Oxygen radicals in the suspension signal strength. Results: The safflower water extract of each dose group can significantly increase the frequency of myocardial cell injury after H2O2 injury, so that cell survival rate, reduce the release of LDH cardiomyocytes, reduce the content of MDA and XOD in culture medium, while increasing the activity of SOD, NO and GSH-Px activity, so that the cell suspension of ESR oxygen free radicals significantly decreased signal. CONCLUSION: The aqueous extract of safflower can protect neonatal rat cardiomyocytes in vitro. The mechanism is related to reducing the production of reactive oxygen species, enhancing the clearance of oxygen free radicals and enhancing the activity of endogenous antioxidant enzymes.