摘要
目的研究小檗碱对缺氧复氧心肌细胞损伤的影响。方法构建缺氧复氧心肌细胞损伤模型。将心肌细胞分成正常组(正常培养)、模型组(缺氧复氧处理)、小檗碱组(缺氧复氧处理前1h,添加50μmol/L的小檗碱预处理)、小檗碱+PMA组(缺氧复氧处理前1h,添加50μmol/L的小檗碱和1μmol/L的NF-κB信号激活剂PMA预处理),免疫印迹检测各组细胞中核因子-κBp65(NF-κBp65)蛋白表达变化。用小檗碱及联合NF-κB信号激活剂处理缺氧复氧心肌细胞,MTT检测细胞增殖,二硝基苯肼显色法检测LDH漏出率,Annexin V-FITC/PI双染法测定细胞凋亡,免疫印迹检测Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平,黄嘌呤氧化法检测SOD活性,硫代巴比妥酸法检测MDA含量。结果缺氧复氧处理后的心肌细胞中NF-κBp65蛋白水平升高,细胞增殖活性降低,LDH漏出率升高,细胞凋亡率升高,细胞中Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平升高,SOD活性降低,MDA含量升高。小檗碱可以降低缺氧复氧条件下心肌细胞中NF-κBp65蛋白表达水平,提高细胞增殖活性,降低LDH漏出率,减少细胞凋亡和细胞中Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平,提高SOD活性并减少MDA含量。NF-κB信号激活剂处理可以逆转小檗碱对缺氧复氧心肌细胞增殖、LDH漏出率、凋亡及SOD、MDA水平的影响。结论小檗碱通过下调NF-κB信号激活水平抑制缺氧复氧心肌细胞损伤。
Objective To investigate the effect of Berberine on hypoxia-reoxygenation induced injury of cardiomyocytes. Methods The models of cardiomyocytes with hypoxia-reoxygenation induced injury were established. The cardiomyocytes were divided into the normal group (normal culture), model group (hypoxia-reoxygenation treatment), Berberine group (1 h before hypoxia-reoxygenation treatment, pretreatment with 50 μmol/L Berberine), Berberine+PMA group (pretreatment with 50 μmol/L Berberine and 1 μmol/L nuclear factor-κB (NF-κB) signal activator PMA at 1 h before hypoxia-reoxygenation treatment). Western blotting was used to determine the change of protein expression level of NF-κ Bp65 in each group. The Berberine and NF-κB signal activator were used to treat cardiomyocytes with hypoxia-reoxygenation induced injury. MTT was used to examine cell proliferation. Dinitrophenyl hydrazine colorimetry was used to examine the leakage rate of LDH. Annexin V-FITC/PI double staining was used to determine cell apoptosis. The protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were determined by Western blotting. Xanthine oxidation was used to determine SOD activity. Thiobarbituric acid assay was used to examine the MDA level. Results The protein level of NF-κB p65 in cardiomyocytes treated with hypoxia-reoxygenation increased, the activity of cell proliferation decreased, the LDH leakage rate increased, the apoptosis rate increased, the protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 increased, the SOD activity decreased, and the MDA level increased. Berberine could reduce the protein expression level of NF-κB p65 in cardiomyocytes under hypoxia-reoxygenation, increase activity of cell proliferation, decrease LDH leakage rate, reduce cell apoptosis and protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9, increase SOD activity, and reduce MDA level. The treatment with NF-κB signal activator could reverse the effect of Berberine on the proliferation of cardiomyocytes with hy
作者
张丽平
毛蕾芳
孙玉梅
Zhang Liping;Mao Leifang;Sun Yumei(Department of Cardiac Surgery,Henan Chest Hospital,Zhengzhou,Henan 450000,China;Pain Department,Henan Provincial Chest Hospital,Zhengzhou,Henan 450000,China;Department of Cardiology,Henan Chest Hospital,Zhengzhou,Henan 450000,China)
出处
《中华生物医学工程杂志》
CAS
2019年第1期57-62,共6页
Chinese Journal of Biomedical Engineering
基金
2017年河南省医学科技攻关项目(201702293).
