生理病理状态下大鼠体内荭草花抗心肌缺血再灌注损伤活性成分筛选
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篇名: 生理病理状态下大鼠体内荭草花抗心肌缺血再灌注损伤活性成分筛选
TITLE: Screening of active components of Polygonum orientale flower against myocardial ischemia-reperfusion injury in rats under physiological and pathological states
摘要: 目的 基于生理病理状态筛选大鼠体内荭草花抗心肌缺血再灌注损伤(MIRI)的潜在活性成分。方法将SD大鼠分为正常对照组、正常给药组、MIRI对照组和MIRI给药组,每组5只。经药物干预或造模、药物干预后,收集各组大鼠的血浆样品,使用超高效液相色谱-四极杆-飞行时间质谱联用技术进行色谱分离和质谱数据采集,经比对对照品图谱、各血浆样品图谱并参考相关文献后,对原型入血成分和代谢产物进行分析;同时,确定正常给药组和MIRI给药组大鼠血浆样品中的共有峰,结合主成分分析和“有监督”的正交偏最小二乘法-判别分析,以变量重要性投影(VIP)值>1为标准,筛选差异移行成分,并推测生理病理状态下荭草花抗MIRI的潜在活性成分。将SD大鼠分为对照组、MIRI组、阳性对照组(复方丹参片0.2g/kg,每天3次)、各潜在活性成分组(各潜在活性成分均为10mg/kg,每天2次),每组5只。各药物组大鼠灌胃相应药液,连续3d。末次给药后,对各组大鼠血浆中超氧化物歧化酶(SOD)活性和乳酸脱氢酶(LDH)、肌酸激酶同工酶MB(CK-MB)、心肌肌钙蛋白Ⅰ(cTnⅠ)泄漏量进行检测、比较。结果从荭草花提取物总离子流图中获取主要色谱峰26个,确定了其中14个色谱峰,包括没食子酸、儿茶素、原儿茶酸等。从正常大鼠血浆样品中检测到移行成分15个,其中原型入血成分6个、代谢产物9个;从MIRI大鼠血浆样品中检测到移行成分19个,其中原型入血成分6个、代谢产物13个。两者共有移行成分8个,其中山柰酚的葡萄糖醛酸化代谢产物、槲皮素的羰基化代谢产物和N-p-香豆酰酪胺对应色谱峰的VIP值均大于1。与MIRI组比较,各潜在活性成分组大鼠血浆SOD活性均显著升高(P<0.01),LDH、CK-MB、cTnⅠ泄漏量均显著降低(P<0.01)。结论N-p-香豆酰酪胺、槲皮素、山柰酚和山柰素-3-O-β-D-葡萄糖苷可能是荭草花提取物抗MIRI的潜在活性成分。
ABSTRACT: OBJECTIVE To screen the potential active components of Polygonum orientale flower against myocardial ischemia- reperfusion injury (MIRI) in rats based on physiological and pathological states. METHODS SD rats were divided into normal control group, normal administration group, MIRI control group and MIRI administration group, with 5 rats in each group. After drug intervention or modeling and drug intervention, chromatographic separation plasma samples were collected, and chromatographic separation and mass spectrometry data collection were performed by using UPLC-Q-TOF/MS. The prototype components and metabolites were analyzed by comparing the reference substance maps, the maps of each plasma sample, and the relevant literature. At the same time, the common peaks in plasma samples of rats in normal administration group and MIRI administration group were identified. Combined with principal component analysis and orthogonal partial least square-discriminant analysis, the differential transitional components were screened out according to the value of variable importance in the projection (VIP)>1, to speculate the potential active components of P. orientale flower in rats under physiological and pathological states. The SD rats were divided into control group, MIRI group, positive control group (Compound danshen tablets 0.2 g/kg, 3 times a day), and potentially active compound groups (10 mg/kg, twice a day), with 5 rats in each group. The rats in administration groups were given relevant medicine intragastrically, for 3 consecutive days. The activity of superoxide dismutase (SOD), the leakages of lactate dehydrogenase (LDH), creatine kinase isoenzyme-MB (CK-MB) and cardiac troponin Ⅰ (cTnⅠ) in plasma were detected after the last administration. RESULTS Twenty-six main chromatographic peaks were obtained from the total ion chromatogram of the extract of P. orientale flower, and 14 of them were determined, including gallic acid, catechin, protocatechuic acid and so on. There were fifteen (including 6 absorbed prototype components and 9 metabolites) and nineteen transitional components (including 6 absorbed prototype components and 13 metabolites) in the plasma sample of normal rats and MIRI rats. Eight transitional components were detected in both normal rats and MIRI rats, and the VIP values of kaempferol glucuronidation metabolites, quercetin carbonylation metabolites and N-p-paprazine to the corresponding peak were higher than 1. Compared with MIRI group, the activities of SOD were increased significantly in the plasma of MIRI rats in each potential active compound group (P<0.01), and the leakages of LDH, CK-MB, and cTnⅠ in the plasma of MIRI rats were reduced significantly (P<0.01). CONCLUSIONS The potential anti-MIRI active components in extract of P. orientale flower are N-p-paprazine, quercetin, kaempferol and kaempferol-3-O-β-D-glucoside.
期刊: 2024年第35卷第16期
作者: 仁莎莎;胡建春;张元现;陈轻轻;刘春花;郑林;巩仔鹏;黄勇;金阳;李月婷
AUTHORS: REN Shasha,HU Jianchun,ZHANG Yuanxian, CHEN Qingqing,LIU Chunhua,ZHENG Lin,GONG Zipeng,HUANG Yong,JIN Yang,LI Yueting
关键字: 荭草花;心肌缺血再灌注损伤;活性成分;血浆药物化学
KEYWORDS: Polygonum orientale flower; myocardial ischemia-reperfusion injury; active ingredients; plasma pharmacochemistry
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