樱黄素对心肌梗死模型小鼠的治疗作用及其机制

doi:10.13481/j.1671-587X.20260113

摘要/Abstract

摘要：

目的探讨樱黄素对心肌梗死（MI）小鼠的治疗作用，并阐明樱黄素治疗MI的关键靶点及分子机制。方法从50只c57小鼠中随机选取10只小鼠作为假手术组；另外40只小鼠采用心脏左前降支结扎建立小鼠MI模型，将造模成功的40只小鼠随机分为模型组、低剂量樱黄素组（5 mg·kg^-1樱黄素）、高剂量樱黄素组（10 mg·kg^-1樱黄素）和阳性药物组（依那普利组）（2 mg·kg^-1依那普利），每组10只。持续21 d腹腔注射给药，每日1次。采用超声心动图检测各组小鼠心功能指标，酶联免疫吸附试验（ELISA）法检测各组小鼠血清中肌酸激酶同工酶（CK-MB）和肌钙蛋白Ⅰ（cTn-Ⅰ）水平，HE染色观察各组小鼠心肌组织病理形态表现，Masson染色观察各组小鼠心肌组织纤维化情况，免疫组织化学染色法和免疫荧光法检测各组小鼠MI边缘区心肌组织中CD31表达情况及新生血管数量。利用有机小分子生物活性数据库（Pubchem）获取樱黄素结构信息，采用蛋白质序列数据库（SWISS）和定量构效关系数据库（QSAR）获取樱黄素的靶点，采用疾病基因网络数据库（GeneCards）、基因网络数据库（Disgene）和在线人类孟德尔遗传数据库（OMIM）获取MI疾病靶点，搜索关键词“心肌梗死”，获得与之相关的全部疾病靶点。将樱黄素的靶点与疾病的靶点进行交集匹配并寻找共同靶点。运用String数据库构建蛋白-蛋白相互作用（PPI）网络图。选取排名靠前的靶点与樱黄素进行分子对接，分析樱黄素治疗MI的信号通路。结果与假手术组比较，模型组小鼠左心室射血分数（LVEF）和左心室短轴缩短率（LVFS）明显降低（P<0.001）；与模型组比较，低和高剂量樱黄素组及依那普利组小鼠LVEF和LVFS均明显升高（P<0.05）。ELISA法，与假手术组比较，模型组小鼠血清中CK-MB和cTn-Ⅰ水平均明显升高（P<0.001）；与模型组比较，低和高剂量樱黄素组及依那普利组小鼠血清中CK-MB和cTn-Ⅰ水平均明显降低（P<0.01）。HE染色，模型组小鼠心肌细胞形态改变，组织排列紊乱并且有明显断裂，炎性细胞浸润增多；低剂量樱黄素组小鼠心肌细胞坏死程度有所减轻；高剂量樱黄素组小鼠心肌细胞的肿胀、排列失序和肌纤维断裂的症状明显缓解。Masson染色，模型组小鼠心肌纤维化明显，心室前壁变薄，而低剂量樱黄素组小鼠心肌纤维化面积降低，显著抑制纤维化的发展，高剂量樱黄素组小鼠心肌纤维化面积明显减少，纤维化导致的瘢痕样改变减轻。免疫组织化学染色法和免疫荧光法，与模型组比较，高剂量樱黄素组小鼠MI边缘区新生血管数量升高（P<0.05）。网络药理学，樱黄素化合物靶点100个，其中68个为樱黄素-MI的共同靶点。与MI有关的关键基因包括表皮生长因子受体（EGFR）、磷脂酰肌醇3-激酶催化亚单位α（PIK3CA）和过氧化物酶体增生激活受体γ（PPARG）等。分子对接分析结果显示樱黄素与关键靶点EGFR和PIK3CA对接良好，与EGFR的亲和性更好。结论樱黄素可促进MI小鼠模型梗死边缘区毛细血管新生以减轻缺血缺氧损伤，从而减轻MI后的小鼠心功能障碍。

关键词: 心肌梗死, 樱黄素, 血管再生, 网络药理学, 分子对接

Abstract:

Objective To discuss the therapeutic effect of prunetin on the mice with myocardial infarction （MI）， and to clarify the key targets and molecular mechanism of prunetin in the treatment of MI. Method Ten mice were randomly selected from 50 c57 mice as sham operation group； the other 40 mice were used to establish the MI mouse models by ligating the left anterior descending coronary artery. The 40 successfully modeled mice were randomly divided into model group， low dose of prunetin group （5 mg·kg?1 prunetin）， high dose of prunetin group （10 mg·kg?1 prunetin）， and positive drug group （2 mg·kg?1 enalapril）， with 10 mice in each group. The drugs were administered by intraperitoneal injection once daily for 21 consecutive days. Echocardiography was used to detect the cardiac function indexes of the mice in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of creatine kinase-MB （CK-MB） and cardiac troponin Ⅰ （cTn-Ⅰ） in serum of the mice in various groups； HE staining was used to observe the patho morphology of myocardium tissue of the mice in various groups； Masson staining was used to observe the fibrosis of myocardium tissue of the mice in various groups； immunohistochemistry and immunofluorescence methods were used to detect the expression of CD31 and the number of new blood vessels in myocardium tissue at the MI border zone of the mice in various groups. The Pubchem database was used to obtain the structural information of prunetin； the SWISS and QSAR databases were used to obtain the targets of prunetin； the The GeneCards Human Gene Database （GeneCards）， Gene Web database（Disgene） and Online Mendelian Inheritance in Man （OMIM） database were used to obtain the disease targets of MI； the targets of prunetin and the disease targets were intersected to identify common targets. The String database was used to construct the protein-protein interaction （PPI） network diagram. The top-ranked targets were selected for molecular docking with prunetin to analyze the signaling pathways of prunetin in the treatment of MI. Result The echocardiography results showed that compared with sham operation group， the left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） of the mice in model group were significantly decreased （P<0.001）； compared with model group， the LVEF and LVFS of the mice in low dose and high doses of prunetin groups and enalapril group were significantly increased （P<0.05）. The ELISA results showed that compared with sham operation group， the serum levels of CK-MB and cTn-Ⅰ of the mice in model group were increased （P<0.001）； compared with model group， the levels of CK-MB and cTn-Ⅰ in the serum of the mice in low and high doses of prunetin groups and enalapril group were significantly decreased （P<0.01）. The HE staining results showed that in model group， the morphology of myocardial cells was altered， the tissue arrangement was disordered with obvious rupture， and inflammatory cell infiltration was increased； in low dose of prunetin group， the degree of myocardial cell necrosis was alleviated； in high dose of prunetin group， the symptoms of myocardial cell swelling， disordered arrangement， and myofibril rupture were significantly relieved. The Masson staining results showed that in model group， myocardial fibrosis was obvious and the anterior ventricular wall was thinned， while in low dose of prunetin group， the myocardial fibrosis area was reduced， significantly inhibiting the development of fibrosis， and in high dose of prunetin group， the myocardial fibrosis area was significantly decreased， and the scar-like changes caused by fibrosis were alleviated. The immunohistochemistry and immunofluorescence results showed that compared with model group， the number of new blood vessels in the MI border zone myocardium tissue of the mice in high dose of prunetin treatment group was increased （P<0.05）. The network pharmacology results showed that there were 100 compound targets of prunetin， among which 68 were common targets for prunetin-MI. The key genes related to MI included epidermal growth factor receptor （EGFR）， phosphatidylinositol 3-kinase catalytic subunit alpha（PIK3CA）， and peroxisome proliferator-activated receptor gamma（PPARG）， etc. The molecular docking analysis results showed that prunetin docked well with the key targets EGFR and PIK3CA， and had better affinity with EGFR. Conclusion Prunetin can promote the angiogenesis of capillaries in the infarct border zone of the MI mouse models to alleviate ischemia-hypoxia injury， thereby alleviating cardiac dysfunction in mice after MI.

Key words: Myocardial infarction, Prunetin, Angiogenesis, Network pharmacology, Molecular docking

中图分类号:

R542.22

蔡萌,郭杨,马莹芳,崔静蕾,罗佳,魏丽丽,张云华,王洋. 樱黄素对心肌梗死模型小鼠的治疗作用及其机制[J]. 吉林大学学报(医学版), 2026, 52(1): 125-134.

Meng CAI,Yang GUO,Yingfang MA,Jinglei CUI,Jia LUO,Lili WEI,Yunhua ZHANG,Yang WANG. Therapeutic effect of prunetin on myocardial infarction model mice and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 125-134.

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Group	n	LVEF	LVFS
Sham operation	9	75.76±3.34	43.72±3.09
Model	8	36.15±11.33^*	17.47±5.74^*
Low dose of prunetin	8	46.75±4.26^△	23.20±4.08^△
High dose of prunetin	8	52.65±4.97^△	27.32±3.38^△
Enalaprilat	8	59.42±3.52^△	31.10±2.25^△

Group	CK-MB [ρ_B/(μg·L^-1)]	cTn-Ⅰ [ρ_B/(ng·L^-1)]
Sham operation	4.53±2.12	33.33±13.41
Model	23.44±9.53^*	62.42±7.05^*
Low dose of prunetin	7.00±2.30^△	32.46±14.85^△
High dose of prunetin	5.83±1.38^△	28.70±11.70^△
Enalaprilat	6.27±1.37^△	48.83±11.12^△

Group	Number of new vessels
Sham operation	12.24±5.62
Model	10.67±3.74
Low dose of prunetin	18.22±5.26
High dose of prunetin	22.06±8.65^*
Enalaprilat	12.11±6.50