College of Pharmacy, Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
College of Basic Medicine, Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
Biomed Chromatogr. 2022 Nov;36(11):e5475. doi: 10.1002/bmc.5475. Epub 2022 Sep 7.
This study established the fingerprint of Syringa pinnatifolia Hemsl. (SP), analyzed the SP ingredients absorbed into the rats blood, and evaluated its anti-myocardial ischemic effect to provide a scientific basis for the follow-up development and research of SP and lay a foundation for its clinical application using ultra-performance liquid chromatography-Q Exactive-mass spectrometry and GC-MS. Myocardial infarction was induced in rat by ligating the left anterior descending branch of the rat coronary artery, and SP alcohol extract was administered to evaluate its anti-myocardial ischemic effect. We analyzed the SP ingredients absorbed into the rats blood, screened the active compounds, established a database of SP anti-myocardial ischemic targets, and explored the possible mechanism of SP in treating myocardial infarction using bioinformatics. The rats were examined using echocardiography, serum biomarkers were determined, and pathological changes were observed by histopathological examination. TUNEL staining was performed to detect the apoptotic level of cells, and Western blot and quantitative real-time polymerase chain reaction were performed to detect the expression levels of Bcl-2, Bax, and Caspase-3 in heart tissues. In the fingerprint of SP, 24 common peaks were established, and the similarity evaluation results of 10 batches of SP were all >0.9. Ultra-performance liquid chromatography-Q Exactive-mass spectrometry and GC-MS detected 17 active ingredients in the drug-containing serum, including terpenoids, flavonoids, phenols, phenylpropanoids, and phenolic acids, the most abundant of which was resveratrol. Enrichment analysis of SP targets against myocardial ischemia revealed that key candidate targets of SP were significantly enriched in multiple pathways associated with apoptosis. Resveratrol was administered to the successfully modeled rats, and the results showed that the resveratrol group significantly decreased left ventricular end-diastolic diameter and left ventricular end-systolic diameter and significantly increased ejection fraction and fractional shortening in all groups compared with the model group. Resveratrol significantly decreased the levels of creatine kinase isoenzyme and lactate dehydrogenase in serum compared to the model group (P < 0.001). Hematoxylin-eosin staining of rat myocardial tissue showed that all lesions were reduced under microscopic observation in the resveratrol group compared with the model group. Real-time polymerase chain reaction and Western blot results showed that the resveratrol group downregulated the expression of the proapoptotic factor Bax, upregulated the expression of the antiapoptotic factor Bcl-2, and decreased the expression of Caspase-3. The established fingerprints are accurate, reliable, and reproducible and can be used as an effective method for quality control of the herbs. The anti-myocardial ischemia effect of SP is that resveratrol improves cardiac function and inhibits cardiomyocyte apoptosis to protect cardiomyocytes. The present study provides ample evidence for the clinical use of SP, suggesting that this drug has great potential in the treatment of ischemic heart disease.
本研究建立了紫丁香的指纹图谱,分析了紫丁香中被大鼠血液吸收的成分,并评价了其抗心肌缺血作用,为紫丁香的后续开发研究提供了科学依据,为其临床应用奠定了基础,采用超高效液相色谱- Q Exactive- mass 谱和 GC-MS。通过结扎大鼠冠状动脉左前降支诱导大鼠心肌梗死,给予紫丁香醇提取物评价其抗心肌缺血作用。我们分析了被大鼠血液吸收的紫丁香成分,筛选出活性化合物,建立了紫丁香抗心肌缺血靶点数据库,并通过生物信息学探讨了紫丁香治疗心肌梗死的可能机制。通过超声心动图检查大鼠,测定血清标志物,通过组织病理学检查观察病理变化。TUNEL 染色检测细胞凋亡水平,Western blot 和实时定量聚合酶链反应检测心脏组织中 Bcl-2、Bax 和 Caspase-3 的表达水平。在紫丁香的指纹图谱中,建立了 24 个共有峰,10 批紫丁香的相似度评价结果均>0.9。超高效液相色谱-Q Exactive- mass 谱和 GC-MS 检测到药物血清中的 17 种活性成分,包括萜类、黄酮类、酚类、苯丙素类和酚酸类,其中含量最丰富的是白藜芦醇。紫丁香抗心肌缺血靶点的富集分析表明,紫丁香的关键候选靶点在与凋亡相关的多个途径中显著富集。给予成功建模的大鼠白藜芦醇,结果显示,与模型组相比,白藜芦醇组各亚组左心室舒张末期直径和左心室收缩末期直径显著减小,射血分数和缩短分数显著增加。与模型组相比,白藜芦醇组血清肌酸激酶同工酶和乳酸脱氢酶水平显著降低(P<0.001)。大鼠心肌组织苏木精-伊红染色显示,与模型组相比,白藜芦醇组在显微镜下观察到所有病变均减轻。实时聚合酶链反应和 Western blot 结果显示,白藜芦醇组下调促凋亡因子 Bax 的表达,上调抗凋亡因子 Bcl-2 的表达,降低 Caspase-3 的表达。所建立的指纹图谱准确、可靠、重现性好,可作为该草药质量控制的有效方法。紫丁香的抗心肌缺血作用是白藜芦醇通过改善心功能和抑制心肌细胞凋亡来保护心肌细胞。本研究为紫丁香的临床应用提供了充分的证据,表明该药物在缺血性心脏病治疗中具有很大的潜力。