1. 河南中医药大学第一附属医院
2. 河南中医药大学第一临床医学院
纸质出版:2019
移动端阅览
[1]崔琳,王幼平,王新陆,李彬,郝轩轩,谢世阳,高原,王小晓,王贺,朱明军.参附益心颗粒对缺氧诱导的大鼠H_9C_2心肌细胞能量代谢的影响[J].中医杂志,2019,60(10):862-868.
CUI Lin, WANG Youping, WANG Xinlu, et al. Effects of Shenfu Yixin Granules(参附益心颗粒) on Energy Metabolism of Hypoxia-Induced Rats H9C2 Cardiomyocytes[J]. Journal of Traditional Chinese Medicine2019, 60(10): 862-868. DOI: 10.13288/j.11-2166/r.2019.10.013.
[1]崔琳,王幼平,王新陆,李彬,郝轩轩,谢世阳,高原,王小晓,王贺,朱明军.参附益心颗粒对缺氧诱导的大鼠H_9C_2心肌细胞能量代谢的影响[J].中医杂志,2019,60(10):862-868. DOI: 10.13288/j.11-2166/r.2019.10.013.
CUI Lin, WANG Youping, WANG Xinlu, et al. Effects of Shenfu Yixin Granules(参附益心颗粒) on Energy Metabolism of Hypoxia-Induced Rats H9C2 Cardiomyocytes[J]. Journal of Traditional Chinese Medicine2019, 60(10): 862-868. DOI: 10.13288/j.11-2166/r.2019.10.013. DOI:
目的探讨参附益心颗粒治疗慢性心力衰竭的可能作用机制。方法以不同浓度(0. 1、0. 2、0. 3、0. 4、0. 5、0. 6、0. 9、1. 2 mg/ml)的参附益心颗粒处理H
9
C
2
细胞24 h
MTT法检测细胞活性
筛选后续实验的药物浓度。将H
9
C
2
细胞分为正常组(正常培养)、模型组(缺氧6 h)、曲美他嗪组(1×10
-4
mol/L曲美他嗪+缺氧6 h)、辅酶Q10组(1×10
-4
mol/L辅酶Q10+缺氧6 h)及中药低剂量组(0. 25 mg/ml参附益心颗粒+缺氧6h)、中药高剂量组(0. 5 mg/ml参附益心颗粒+缺氧6h)
检测各组心肌细胞ATP含量、底物代谢酶脂肪酸转运酶(FAT)、肉碱脂酰转移酶1 (MCPT-1)、中链脂酰辅酶A脱氢酶(MCAD)、磷酸腺苷活化蛋白激酶(AMPK)、丙酮酸脱氢酶(PDH)、葡萄糖转运子4 (GLUT-4)、腺苷酸转运体(ANT)、肌酸激酶(CK)蛋白表达水平;观察各组H
9
C
2
心肌细胞超微结构的变化。将H
9
C
2
细胞分为正常组、模型组、中药组(0. 5 mg/ml参附益心颗粒+缺氧6 h)
JC-1染色检测线粒体膜电位变化。结果 0. 5 mg/ml参附益心组及低于此浓度组无细胞毒性
以0. 5 mg/ml参附益心颗粒用于中药组。与正常组比较
模型组H
9
C
2
细胞ATP含量及FAT、MCPT-1、MCAD、GLUT-4、ANT、CK蛋白含量明显降低
AMPK、PDH蛋白含量明显升高(P
<
0. 05)。与模型组比较
中药组和辅酶Q10组心肌细胞中ATP含量及FAT、MCPT-1、MCAD、GLUT-4、ANT、CK蛋白含量均明显升高(P
<
0. 05)。正常组细胞线粒体结构正常
模型组细胞膜破裂
线粒体肿胀
边界模糊不清。与模型组对比
中药组细胞损伤明显减轻
细胞膜完整
线粒体结构清晰。JC-1染色显示中药组可以改善线粒体膜电位的下降。结论参附益心颗粒可能通过升高心肌H
9
C
2
细胞ATP、GLUT-4、ANT、FAT、MCPT-1、MCAD、CK蛋白表达
降低AMPK、PDH蛋白表达
改善缺氧诱导H
9
C
2
心肌细胞线粒体膜电位
从而发挥治疗慢性心力衰竭的作用。
Objective To investigate the possible mechanism of Shenfu Yixin Granules( 参附益心颗粒) in treating chronic cardiac failure. Methods H
9
C
2
cells were cultured for 24 h with Shenfu Yixin Granules at different concentrations( 0. 1
0. 2
0. 3
0. 4
0. 5
0. 6
0. 9
1. 2 mg/ml)
and the cell activity was determined by MTT colorimetric assay to screen for drug concentrations in subsequent experiments. The H
9
C
2
cells were divided into normal group( normal culture)
model group( hypoxia 6 h)
trimetazidine group( 1 × 10
-4
mol/L trimetazidine + hypoxia 6 h)
coenzyme Q10 group( 1 × 10
-4
mol/L coenzyme Q10 + hypoxia 6 h)
low dose Chinese medicine group( 0. 25 mg/ml Shenfu Yixin Granules + hypoxia 6 h)
and high dose Chinese medicine group( 0. 5 mg/ml Shenfu Yixin Granules + hypoxia 6 h). The protein expression levels of adenosine triphosphate( ATP)
fatty acid transporter( FAT)
carnitine lipid acetyltransferase 1( MCPT-1)
medium chain Lipoxycoenzyme A dehydrogenase( MCAD)
phosphate activated protein Kinase( AMPK)
pyruvate dehydrogenase( PDH)
glucose transporter 4( GLUT-4)
adenylate transporter( ANT)
creatine kinase( CK) in the cardiomyocytes of each group were detected. The ultrastructure changes of H
9
C
2
cardiomyocytes in each group were observed. H
9
C
2
cells were divided into normal group
model group and Chinese medicine group( 0. 5 mg/ml Shenfu Yixin Granules + hypoxia 6 h). Mitochondrial membrane potential was detected by JC-1 staining. Results There was no cytotoxicity in the 0. 5 mg/ml Shenfu Yixin Granules group and the lower concentration group
which was suitable for administration of 0. 5 mg/ml Shenfu Yixin Granules in Chinese medicine group. Compared with the normal group
the ATP content and FAT
MCPT-1
MCAD
GLUT-4
ANT
CK protein contents in H
9
C
2
cells in the model group were significantly decreased
and AMPK
PDH protein contents were significantly increased( P
<
0. 05). Compared with the model group
the ATP content and FAT
MCPT-1
MCAD
GLUT-4
ANT
CK protein contents in the cardiomyocytes of Chinese medicine group and coenzyme Q10 group were significantly increased( P
<
0. 05). In the normal group
the mitochondria structure was normal. The membrane of the model group was broken
the mitochondria were swollen
and the boundary was blurred. Compared with the model group
the cell damage in the Chinese medicine group was obviously alleviated
the cell membrane was intact
and the mitochondrial structure was clear. JC-1 staining showed that the Chinese medicine group could improve the decrease of mitochondrial membrane potential. Conclusion Shenfu Yixin Granules may increase the expression of ATP
GLUT-4
ANT
FAT
MCPT-1
MCAD
CK protein
decreased the expression of AMPK and PDH protein
improve mitochondrial membrane potential of H
9
C
2
cardiomyocytes induced by hypoxia
so as to play the role for treating chronic cardiac failure.
0
浏览量
506
下载量
7
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010102007558号