Page 52 - 《中国药房》2021年第9期

干姜水煎液对心肌细胞线粒体功能损伤的改善作用研究 Δ

文建霞 ，赵艳玲（中国人民解放军总医院第五医学中心药学部，北京 100039）
#
＊
中图分类号 R285.5 文献标志码 A 文章编号 1001-0408（2021）09-1070-07
DOI 10.6039/j.issn.1001-0408.2021.09.08

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、8 mg/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、0.5 mg/mL）对阿霉素（5 μmol/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（0.5 mg/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、0.5 mg/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、0.5 mg/mL（或
0.5 mg/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。
关键词 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

Study on Improvement Effects of Zingiber officinale Decoction on Mitochondrial Function Injury of
Cardiomyocytes
WEN Jianxia，ZHAO Yanling（Dept. of Pharmacy，Fifth Medical Center of PLA General Hospital，Beijing
100039，China）

ABSTRACT OBJECTIVE：To investigate the improvement effects of Zingiber officinale decoction （ZOD） on doxorubicin
（DOX）-induced mitochondrial function injury of H9c2 cardiomyocytes. METHODS：Taking H9c2 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 H9c2 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 H9c2 cardiomyocytes were increased significantly（P＜0.01）or had no statistical significance
（P＞0.05）. After DOX induced mitochondrial dysfunction of H9c2 cardiomyocytes，pretreated with 0.125，0.25，0.5 mg/mL（or
0.5 mg/mL）ZOD，the morphology of H9c2 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
Δ 基金项目：国家自然科学基金资助项目（No.81573631）；北京市 mitochondrial energy metabolism of H9c2 cardiomyocytes，so
科技计划项目（No.Z171100001717013） as to improve mitochondrial function injury.
＊博士研究生。研究方向：中药基础理论。电话：010-66933224。 KEYWORDS Zingiber officinale decoction； H9c2
E-mail：wenjx32@163.com cardiomyocytes； Mitochondrial function injury； Respiratory
# 通信作者：研究员，博士生导师，博士。研究方向：中药药理与 function；Energy metabolism
新药研发。电话：010-66933320。E-mail：zhaoyl2855@126.com

·1070 · China Pharmacy 2021 Vol. 32 No. 9 中国药房 2021年第32卷第9期