摘要
目的:分析中药单体山萘酚和芒柄花黄素对缺氧条件下H9c2心肌细胞内活性氧水平及NADPH氧化酶亚基NOX4、p67^phox、p47^phox表达量的影响。方法:采用缺氧造模的方法对H9c2心肌细胞进行造模,通过倒置荧光显微镜观察DCFH-DA染色的H9c2心肌细胞的荧光强弱;Real timePCR和Westernblot检测NOX4、p67^phox、p47^phox表达量变化。结果:各组细胞内绿色荧光有显著变化,即对照组的细胞内荧光微弱,模型组荧光强度显著增强,各给药组荧光强度有所减弱,作用与阳性药组相似;山萘酚能够有效降低NOX4、p67^phox、p47^phoxmRNA的表达量,芒柄花黄素能够有效降低NOX4、p47^phoxmRNA的表达量;山萘酚能够有效降低NOX4、p67^phox、p47^phox蛋白的表达量,芒柄花黄素能够有效降低NOX4、p67^phox蛋白的表达量。结论:抑制NOX4、p67^phox、p47^phox的表达量可能是山萘酚和芒柄花黄素发挥抗氧化作用的重要环节,而对于NADPH氧化酶的影响反映了H9c2心肌细胞内活性氧的水平。
Objective:To investigate the effects of kaempferol and formononetin on the expressions of reactive oxygen species(ROS) and NADPH oxidase(NOX4,p67phox and p47phox) inhypoxia H9 c2 cardiomyocytes.Methods:H9 c2 cardiomyocytes were modeled under hypoxia.The fluorescence intensity of H9 c2 cardiomyocytes stained with DCFH-DA was observed by inverted fluorescence microscopy.The expressions of NOX4,p67phox and p47phox were observed by Real time PCR and Western blot.Results:The fluorescence intensity of each group was significantly changed.The fluorescence intensity of control group and each administration group were weak.However,the model group was the opposite.Kaempferol can effectively reduce the mRNA expressions of NOX4,p67phox,p47phox and the protein expressions of NOX4,p67phox and p47phox.Formononetin can effectively reduce the mRNA expressions of NOX4 and p47phox and the protein expressions of NOX4 and p67phox.Conclusion:Kaempferol and formononetin may play an antioxidant effect by inhibiting the expressions of NOX4,p67phox and p47phox.The effect on NADPH oxidase is related to the level of reactive oxygen species in H9 c2 cardiomyocytes.
作者
于雪
郭淑贞
石天娇
王淑艳
范盎然
王旭
王伟
YU Xue;GUO Shuzhen;SHI Tianjiao;WANG Shuyan;FAN Angran;WANG Xu;WANG Wei(College of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《辽宁中医杂志》
CAS
2018年第12期2465-2468,2701,共5页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金青年项目(81403319)
中央高校基本科研业务费专项资金资助项目(2017-JYB-JS-013)