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中国中医科学院 西苑医院基础医学研究所,国家中医心血管疾病临床医学研究中心, 中药药理北京市重点实验室,北京 100091
Received:08 July 2021,
Published Online:28 September 2021,
Published:20 November 2021
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李磊,孟红旭,辛高杰等.双参宁心胶囊对冠状动脉微循环障碍大鼠心脏功能及血流动力学的影响[J].中国实验方剂学杂志,2021,27(22):41-50.
LI Lei,MENG Hong-xu,XIN Gao-jie,et al.Effect of Shuangshen Ningxin Capsules on Cardiac Function and Hemodynamics in Rats with Coronary Microvascular Dysfunction[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(22):41-50.
李磊,孟红旭,辛高杰等.双参宁心胶囊对冠状动脉微循环障碍大鼠心脏功能及血流动力学的影响[J].中国实验方剂学杂志,2021,27(22):41-50. DOI: 10.13422/j.cnki.syfjx.20212105.
LI Lei,MENG Hong-xu,XIN Gao-jie,et al.Effect of Shuangshen Ningxin Capsules on Cardiac Function and Hemodynamics in Rats with Coronary Microvascular Dysfunction[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(22):41-50. DOI: 10.13422/j.cnki.syfjx.20212105.
目的
2
研究益气活血中药双参宁心胶囊对冠状动脉微循环障碍大鼠心脏血流动力学及心功能的影响。
方法
2
大鼠随机分为6组,分别为假手术组,模型组,尼可地尔组(5 mg·kg
-1
),双参宁心胶囊高、中、低剂量(180,90,45 mg·kg
-1
)组。动物按分组先给予相应药物7 d,末次给药后2 h进行模型制备。采用左心室注射内注射栓塞微球(40~120 μm,约1 000个微球)方法建立大鼠冠脉微循环障碍模型。造模术后24 h后超声心动图观察左室舒张末期内径(LVIDd),左室收缩末期内径(LVIDs),左室舒张末期容积(LVEDV),左室收缩末期容积(LVESV),每搏输出量(SV),心输出量(CO),左室射血分数(EF),左室缩短率(FS);心导管技术测定大鼠动脉收缩压(SBP),舒张压(DBP),左室峰压(LVSP),左室舒张末压(LVEDP),左室内压最大上升速率(LV+dp/dt
max
),左室内压最大下降速率(LV-dp/dt
max
),计算平均动脉(MAP),体表标准Ⅱ导心电图计算心率(HR);生化分析法检测肌酸激酶(CK),肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)活性;酶联免疫吸附测定法(ELISA)检测血清心肌肌钙蛋白T(cTnT);蛋白免疫印迹法(Western blot)检测大鼠心肌组半胱氨酸天冬氨酸蛋白水解酶-3(Caspase-3),B细胞淋巴瘤-2(Bcl-2)及Bcl-2相关X蛋白(Bax)蛋白表达;2,3,5-氯化三苯基四氮唑(TTC)染色观察心肌梗死面积;苏木素-伊红(HE)染色观察心肌形态学改变。
结果
2
与假手术组比较,模型组SV,CO,EF,FS显著降低(
P
<
0.01),LVIDs,LVEDV显著增加(
P
<
0.01);与模型组比较,双参宁心胶囊各剂量组可以增加EF(
P
<
0.05,
P
<
0.01)和FS(
P
<
0.01);180,90 mg·kg
-1
剂量组明显降低LVIDs,LVESV,增加LVEDV,SV和CO(
P
<
0.05,
P
<
0.01)。与假手术组比较,模型组SBP,DBP,MAP,LVSP,LV+dp/dt
max
及LV-dp/dt
max
均显著降低(
P
<
0.01),HR差异无统计学意义。双参宁心胶囊各剂量组对大鼠血流动力学具有一定程度改善,较模型组增加大鼠SBP,DBP,MAP,LVSP,LV+dp/dt
max
,LV-dp/dt
max
及HR(
P
<
0.05,
P
<
0.01)。与假手术组比较,模型组大鼠血清CK,LDH,CK-MB及cTnT明显升高(
P
<
0.01),与模型组比较,双参宁心胶囊有明显降低动物血清CK,LDH,CK-MB及cTnT的作用(
P
<
0.05,
P
<
0.01)。与假手术组比较,模型组大鼠心肌Caspase-3蛋白表达显著升高(
P
<
0.01),Bcl-2蛋白表达明显降低(
P
<
0.05)。与模型组比较,双参宁心胶囊低剂量组心肌Caspase-3蛋白表达降低(
P
<
0.05);与模型组比较,双参宁心胶囊高剂量组显著增加大鼠心肌Bcl-2表达(
P
<
0.01)。心肌TTC染色显示,与模型组比较,双参宁心胶囊各剂量组均能显著减少心肌梗死面积(
P
<
0.01)。HE病理结果显示,双参宁心胶囊各剂量组心肌组织病理变化与模型组比较有不同程度的减轻。
结论
2
益气活血中药双参宁心胶囊可以明显增加微球栓塞导致的冠脉微循环障碍的心脏功能,改善心脏血流动力学指标,减少心肌梗死面积,降低CK,LDH,CK-MB及cTnT等心肌损伤标志物水平,其作用机制与抑制心肌细胞凋亡起到保护心肌的作用有关。
Objective
2
To investigate the effect of Shuangshen Ningxin capsules (SSNX) on cardiac hemodynamics and cardiac function in rats with coronary microvascular dysfunction.
Method
2
Rats were randomly divided into a sham operation group, a model group, a nicorandil group (5 mg·kg
-1
), and high- (180 mg·kg
-1
), medium- (90 mg·kg
-1
), and low-dose (45 mg·kg
-1
) SSNX groups. Rats received corresponding drugs for 7 days. Two hours after the last administration, the model of coronary microvascular dysfunction was induced by left ventricular injection of embolic microspheres (40-120 μm, about 1 000 microspheres). Twenty-four hours after modeling, left ventricular internal dimension in diastole (LVIDd), left ventricular internal dimension in systole (LVIDs) left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), stroke volume (SV), cardiac output (CO), left ventricular ejection fraction (EF), and left ventricular shortening rate (FS) were detected by echocardiography. Cardiac catheterization was used to observe the arterial systolic blood pressure (SBP), diastolic blood pressure (DBP), left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), maximum rate of increase in left ventricular pressure (LV+dp/dt
max
), and maximum rate of decrease in left ventricular pressure (LV-dp/dt
max
), and the mean arterial pressure (MAP) was calculated. Heart rate (HR) was calculated according to Ⅱ lead ECG. Biochemical analysis was carried out to detect the activities of creatine kinase (CK), creatine kinase-MB (CK-MB), and lactate dehydrogenase (LDH). Enzyme-linked immunosorbent assay (ELISA) was used to detect serum cardiac troponin T (cTnT). Western blot was used to detect the protein expression of Caspase-3, Bcl-2, and Bax, and 2,3,5-triphenyltetrazolium chloride (TTC) staining to observe the area of myocardial infarction. Hematoxylin-eosin (HE) staining was used to observe the morphological changes of the myocardium.
Result
2
As revealed by echocardiography, compared with the sham operation group, the model group showed reduced SV, CO, EF, and FS (
P
<
0.01), and increased LVIDs and LVEDV (
P
<
0.01). Compared with the model group, the SSNX groups showed increased EF (
P
<
0.05,
P
<
0.01) and FS (
P
<
0.01), and the high- and medium-dose SSNX groups displayed reduced LVIDs and LVESV, and increased LVEDV, SV, and CO (
P
<
0.05,
P
<
0.01). SBP, DBP, MAP, LVSP, LV+dp/dt
max
, and LV-dp/dt
max
in the model group were lower than those in the sham operation group (
P
<
0.01), while there was no significant difference in HR. SSNX improved hemodynamics of rats, and increased SBP, DBP, MAP, LVSP, LV+dp/dt
max
, LV-dp/dt
max
, and HR as compared with the model group (
P
<
0.05,
P
<
0.01). The serum CK, LDH, CK-MB, and cTnT levels in the model group were higher than those in the sham operation group (
P
<
0.01). Compared with the model group, SSNX groups reduced serum CK, LDH, CK-MB, and cTnT (
P
<
0.05,
P
<
0.01). Compared with the sham operation group, the model group displayed increased expression of Caspase-3 protein in the myocardium (
P
<
0.01) and reduced expression of Bcl-2 protein (
P
<
0.05). The expression of Caspase-3 protein in the myocardium of SSNX groups was lower than that in the model group, and statistical difference was observed between the low-dose SSNX group and the model group (
P
<
0.05). Compared with the model group, the SSNX groups exhibited increased expression of Bcl-2 in the rat myocardium, and the statistical difference was observed in the high-dose SSNX group
(P
<
0.01). As demonstrated by the TTC staining, compared with the model group, SSNX groups showed reduced areas of myocardial infarction (
P
<
0.01). The HE staining indicated that the pathological injury in myocardial tissues of the SSNX groups was relieved as compared with that in the model group.
Conclusion
2
SSNX can significantly enhance the cardiac function after coronary microvascular dysfunction caused by embolic microspheres, improve cardiac hemodynamics, reduce the area of myocardial infarction, and decrease CK, LDH, CK-MB, and cTnT levels. The mechanism may be related to the inhibition of cardiomyocyte apoptosis to protect the myocardium.
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