干姜水煎液对心肌细胞线粒体功能损伤的改善作用研究
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篇名: 干姜水煎液对心肌细胞线粒体功能损伤的改善作用研究
TITLE:
摘要: 目的:探讨干姜水煎液对阿霉素致H9c2心肌细胞线粒体功能损伤的改善作用。方法:以大鼠H9c2心肌细胞为研究对象,采用CCK-8法检测不同浓度干姜水煎液(0.125、0.25、0.5、1、2、4、8mg/mL,以生药量计,下同)对其存活率的影响;采用高内涵活细胞成像系统检测低、中、高浓度干姜水煎液(0.125、0.25、0.5mg/mL)对阿霉素(5μmol/L)致H9c2心肌细胞线粒体功能损伤后细胞形态学的影响,并对其相对细胞总数、相对活细胞荧光强度、相对死细胞荧光强度进行定量分析;采用生物能量分析仪检测干姜水煎液(0.5mg/mL)对H9c2心肌细胞线粒体功能损伤后线粒体呼吸功能相关指标(耗氧率、细胞外酸化率、基线耗氧率、基线细胞外酸化率、应激耗氧率和应激细胞外酸化率)和能量代谢相关指标(基础呼吸水平、最大呼吸水平、ATP产生水平、H+质子渗漏水平、备用呼吸能力和非线粒体呼吸水平)的影响。结果:经0.125、0.25、0.5mg/mL的干姜水煎液作用后,H9c2心肌细胞存活率显著升高(P<0.01)或差异无统计学意义(P>0.05)。阿霉素致H9c2心肌细胞线粒体功能损伤后,经0.125、0.25、0.5mg/mL(或0.5mg/mL)的干姜水煎液干预,细胞形态恢复正常,呈规则纤维状贴壁分布;相对细胞总数、活细胞荧光强度、耗氧率、细胞外酸化率、基线耗氧率、基线细胞外酸化率、应激耗氧率、应激细胞外酸化率、基础呼吸水平、最大呼吸水平、ATP产生水平、备用呼吸能力和非线粒体呼吸水平均显著升高(P<0.05或P<0.01),相对死细胞荧光强度、H+质子渗漏水平均显著降低(P<0.01)。结论:干姜水煎液可通过提高H9c2心肌细胞线线粒体呼吸功能和能量代谢,进而改善其粒体功能损伤。
ABSTRACT: OBJECTIVE:To investigate the improvement e ffects of Zingiber officinale decoction (ZOD) on doxorubicin (DOX)-induced mitochondrial function injury of H 9c2 cardiomyocytes. METHODS :Taking H 9c2 cardiomyocytes as research object,the effects of different concentrations of ZOD (0.125,0.25,0.5,1,2,4,8 mg/mL,by crude drug ,the same below )on its survival rate were investigated by CCK- 8 assay. The effects of low ,medium and high concentrations of ZOD (0.125,0.25,0.5 mg/mL)on the morphology of H 9c2 cardiomyocytes after DOX (5 μmol/L)induced mitochondrial dysfunction were detected by high content living cell imaging system. The relative number of cells ,the relative fluorescence intensity of living cells and the relative fluorescence intensity of dead cells were analyzed quantitatively. The effects of ZOD (0.5 mg/mL)on related indexes of mitochondrial respiratory function (oxygen consumption rate ,extracellular acidification rate ,baseline oxygen consumption rate , baseline extracellular acidification rate ,stress oxygen consumption rate and stress extracellular acidification rate ) and energy metabolism(basic respiration level ,maximum respiration level ,ATP production level ,H+ proton leakage level ,spare respiration level and non-mitochondrial respiration level )were detected by bioenergy analyzer. RESULTS :After treated with 0.125,0.25,0.5 mg/mL ZOD ,the survival rate of H 9c2 cardiomyocytes were increased significantly (P<0.01)or had no statistical significance (P>0.05). After DOX induced mitochondrial dysfunction of H 9c2 cardiomyocytes,pretreated with 0.125,0.25,0.5 mg/mL(or 0.5 mg/mL)ZOD,the morphology of H 9c2 cardiomyocytes returned to normal and showed regular fibrous adherent distribution. The relative cell number ,fluorescence intensity of living cells ,oxygen consumption rate ,extracellular acidification rate ,baseline oxygen consumption rate ,baseline extracellular acidification rate ,stress oxygen consumption rate ,stress extracellular acidification rate,basic respiration level ,maximal respiration level ,ATP production level ,spare respiration level and non-mitochondrial respiration level were all significantly increased (P<0.05 or P<0.01),while relative dead cell fluorescence intensity and H + proton leakage level were significantly decreased (P<0.01). CONCLUSIONS : ZOD can improve the respiratory function and mitochondrial energy metabolism of H 9c2 cardiomyocytes,so as to improve mitochon drial function injury.
期刊: 2021年第32卷第09期
作者: 文建霞,赵艳玲
AUTHORS:
关键字: 干姜水煎液;H9c2心肌细胞;线粒体功能损伤;呼吸功能;能量代谢
KEYWORDS: Zingiber officinale decoction; H9c2
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