天山花楸叶黄酮提取物抗心肌缺血再灌注损伤作用机制研究

Effect of flavones of Sorbus tianschanica against myocardial ischemia reperfusion injury

目的采用网络药理学方法,筛选天山花楸Sorbus tianschanica叶抗心肌缺血再灌注损伤(MIRI)的活性成分及主要靶点,验证天山花楸叶黄酮化合物对离体心脏缺血再灌注损伤的保护作用及其作用机制。方法通过TCMSP等数据库辅助,筛选出天山花楸叶中具有保护MIRI作用的化合物;构建化合物-靶点-疾病网络及蛋白互作网络,定位关键靶点;建立Langendorff离体大鼠MIRI模型,给予黄酮提取物高剂量组(1.5 mg/L)、低剂量组(0.5 mg/L)与4'-chlorodiazepam(CDZ,1μmol/L)预处理,观察缺血再灌注过程中心功能指标的变化情况,对新鲜的心肌组织进行TTC染色,测定心肌梗死面积;石蜡包埋切片HE染色;心肌组织超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)、还原性谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)、总抗氧化能力(T-AOC)和mPTP孔的开放程度。结果网络药理学方法筛选得到天山花楸抗心肌缺血再灌注损伤活性成分主要为黄酮类化合物,核心靶点为TSPO,与线粒体膜通透性转换孔mPTP密切相关;各给药组均能显著增强心功能指标,减小心肌梗死面积(P<0.01),减轻心肌组织病理损伤,增强心肌组织抗氧化能力(P<0.01),抑制mPTP孔的开放(P<0.01),减轻线粒体水肿;其中,高剂量组作用最强。结论天山花楸叶黄酮提取物可增强心肌舒缩功能,减小心肌梗死面积,减轻心肌组织氧化应激损伤,通过作用于TSPO靶点,抑制mPTP孔开放,减轻心肌细胞线粒体水肿从而保护心肌组织。

Objective To screen active components and main target molecules from Sorbus tianschanica for treatment of myocardial ischemia/reperfusion by network pharmacology methods,and verify its protective effects and mechanism of flavonoids in S.tianschanica(FST).Methods The compounds from S.tianschanica with protective effects against myocardial ischemia reperfusion injury(MIRI)were screened and the compound-target-disease network and protein interaction network were constructed,and the key targets were located through TCMSP database.Langendorff-reperfusion rat isolated-hearts were prepared to induce MIRI.The high dose(1.5 mg/L),low dose(0.5 mg/L)of FST,and 4'-chlorodiazepam(CDZ,1μmol/L)were administrated for pretreatment respectively.Hemodynamic index,infarct size of myocardial tissue,and the pathological changes of myocardium were analyzed.Moreover,superoxide dismutase(SOD),malondialdehyde(MDA),glutathione/oxidized glutathione(GSH/GSSG),catalase(CAT),total antioxidant capacity(T-AOC)and the opening degree of mPTP were determined.Results The active components screened from S.tianschanica by network pharmacology in treatment of myocardial ischemia reperfusion were flavonoids;The core target was TSPO,which was closely related to mitochondrial permeability transition pore mPTP.Each treatment group significantly enhanced the cardiac function index,reduced infarct size of myocardial tissue,increased the activity of SOD and CAT(P<0.01),increased GSH/GSSG and T-AOC of myocardial tissue(P<0.01),and inhibited the opening of mPTP.High dose(1.5 mg/L)had the strongest effect by contrast.Conclusion Flavonoids of S.tianschanica can enhance the function of myocardial relaxation and contraction,reduce the area of myocardial infarction and the oxidative stress injury of myocardial tissue;It can inhibit the opening of mPTP by the main target TSPO to reduce the mitochondrial swelling of myocardial cells.