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1.辽宁师范大学 生命科学学院,辽宁 大连 116000
2.中国中医科学院 中药研究所,北京 100075
李硕,在读硕士,从事中药药理毒理研究,E-mail:1137229804@qq.com
叶祖光,博士生导师,研究员,从事中药药理毒理研究,Tel: 010-84252805,E-mail:zgye@icmm.ac.cn
收稿日期:2018-08-10,
网络出版日期:2018-10-19,
纸质出版日期:2019-01-05
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李硕, 苏萍, 张广平, 等. 人参四逆汤及其有效成分对戊巴比妥钠所致心肌细胞损伤模型的保护作用[J]. 中国实验方剂学杂志, 2019,25(1):90-95.
Shuo LI, Ping SU, Guang-ping ZHANG, et al. Protective Effect of Renshen Sinitang and Its Active Ingredients on Myocardial Cell Injury Induced by Pentobarbital Sodium[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 90-95.
李硕, 苏萍, 张广平, 等. 人参四逆汤及其有效成分对戊巴比妥钠所致心肌细胞损伤模型的保护作用[J]. 中国实验方剂学杂志, 2019,25(1):90-95. DOI: 10.13422/j.cnki.syfjx.20190124.
Shuo LI, Ping SU, Guang-ping ZHANG, et al. Protective Effect of Renshen Sinitang and Its Active Ingredients on Myocardial Cell Injury Induced by Pentobarbital Sodium[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 90-95. DOI: 10.13422/j.cnki.syfjx.20190124.
目的:
2
探讨人参四逆汤及其有效成分对戊巴比妥钠所致心肌细胞损伤的保护作用及其机制。
方法:
2
H9C2细胞经传代培养后,分别给予人参皂苷Rb
2
0.01,0.1,1 μmol·L
-1
,Re 0.01,0.1,1 μmol·L
-1
,异甘草素(isoliquiritigenin
ISL)20,40,80 μmol·L
-1
,甘草次酸(glycyrrhetinic acid
GA)10,20,40 μmol·L
-1
,人参四逆汤10
100
400 mg·L
-1
,作用4 h,经0.1%的戊巴比妥钠作用30 min后,检测细胞存活率,乳酸脱氢酶(lactate dehydrogenase
LDH),脂质过氧化物丙二醛(malondialdehyde
MDA),Na
+
-K
+
-三磷酸腺苷(ATP)酶,Ca
2+
-ATP酶活性,同时采用实时荧光定量聚合酶链式反应(Real-time PCR)检测过氧化物酶体增殖活化受体共激活因子-1
α
(peroxisome proliferative activated receptor
PGC-1
α
),B淋巴细胞瘤-2相关X蛋白(Bax),半胱氨酸蛋白酶-3(Caspase-3) mRNA的表达情况。
结果:
2
人参四逆汤及其有效成分对心衰细胞模型有保护作用,与正常组比较,模型组细胞存活率显著下降,LDH
MDA含量显著升高,Na
+
-K
+
-ATP酶活性升高,Ca
2+
-ATP酶活性显著降低,PGC-1
α
mRNA表达下调,Bax
Caspase-3 mRNA表达(
P
<
0.01),证明模型成立。与模型组比较,各给药组显著升高细胞存活率,降低LDH
MDA含量,抑制Na
+
-K
+
-ATP酶活性,提高Ca
2+
-ATP酶活性,上调PGC-1
α
mRNA表达,抑制Bax
Caspase-3 mRNA表达(
P
<
0.05
P
<
0.01)。
结论:
2
人参四逆汤及其有效成分对心衰细胞模型有显著保护作用,且其作用机制与抗氧化,改善线粒体能量代谢,抑制线粒体凋亡途径有关。
Objective:
2
To explore the protective effect and mechanisms of Renshen Sinitang and its active ingredients on cardiomyocyte injury induced by pentobarbital sodium.
Method:
2
H9C2 cells were sub-cultured with ginsenoside Rb
2
0.01
0.1
1 μmol·L
-1
Re 0.01
0.1
1 μmol·L
-1
isoliquiritigenin 20
40
80 μmol·L
-1
glycyrrhetinic acid 10
20
40 μmol·L
-1
Renshen Sinitang
10
100
400 mg·L
-1
for 4 h. After treatment with 0.1% of sodium pentobarbital for 30 min
cell viability
lactate dehydrogenase (LDH)
lipid peroxide malondialdehyde (MDA)
Na
+
-K
+
-adenosine triphosphate(ATP)ase
Ca
2+
-ATPase activity
and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect the expressions of peroxisome proliferative activated receptor-1
α
(PGC-1
α
)
B-cell lymphoma-2 associated X protein(Bax) and cysteine aspartate-specific protease-3(Caspase-3) mRNA.
Result:
2
Renshen Sinitang and its active ingredients have a protective effect on heart failure cell model. Compared with the normal group
the cell survival rate of the model group decreased significantly
while the LDH and MDA contents increased significantly
and the Na
+
-K
+
-ATPase activity increased. Ca
2+
-ATPase activity was significantly decreased
PGC-1
α
mRNA expression was down-regulated
Bax and Caspase-3 mRNA expressions indicates the modeling(
P
<
0.01) . Compared with the model group
each administration group showed a significantly increased cell viability
decreased LDH
MDA content
inhibited Na
+
-K
+
-ATPase activity
increased Ca
2+
-ATPase activity
up-regulated PGC-1
α
mRNA expression
and inhibited Bax and Caspase-3 mRNA expression (
P
<
0.05
P
<
0.01).
Conclusion:
2
Renshen Sinitang and its active ingredients have a significant protective effect on heart failure cell model
and its mechanisms of action are related to anti-oxidation
improvement of mitochondrial energy metabolism and inhibition of mitochondrial apoptosis pathway.
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