The Study Of Protective Effect Of The Active Components In Carthami Flos In Myocardial Ischemia-reperfusion Injury

Carthami Flos is a tubular dried flower of the genus Carthamus in the composite family.It was recorded in the chinese materia medica as medicine mainly by flowers,have dredging the channels and invigorate the circulation of blood the function,Primary treatment gynaecological disease,removing stasis to ease pain and Traumatic injuries.Modern pharmacological studies have confirmed that safflower has the effects of cardiovascular and cerebrovascular protection,anti-inflammatory,neuroprotective,anticoagulant and thrombosis prevention.In particular,it showed protective effect in the clinical treatment of cerebral infarction（CI）and myocardial ischemia reperfusion injury（MIRI）.At present,the study of the protective effect of safflower against myocardial ischemia-reperfusion injury is mainly based on the study of hydroxyl Safflower Yellow A（HSYA）,which is a compound of safflower yellow pigments in Safflower.But other compounds in the Safflower of myocardial ischemia-reperfusion injury of protection rarely reported.Therefore,9 batches of Safflower from different producing areas were selected as research objects in this study.By combining the chemical fingerprint of Safflower UPLC with the results of pharmacodynamic indexes of anti-myocardial ischemia-reperfusion injury,the"spectral effect"relationship analysis was carried out,and the chromatographic peaks of the positive correlation between Safflower and anti-myocardial ischemia-reperfusion injury was obtained by biostatistical analysis methods of cluster analysis,grey correlation analysis,and partial least squares regression analysis.Combined with the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry（UPLC-Q-TOF-MS）technique,the chromatographic peak results were used to infer the potential active components of Safflower that have protective effects on myocardial ischemia-reperfusion injury,which also laid a foundation for the establishment of its quality evaluation system.The experimental results are as follows:（1）The fingerprints of 9 batches of Safflower from different producing areas were obtained by UPLC,and 47 chromatographic peaks were obtained.（2）In vitro hypoxia-reoxygenation injury model was used to simulate ischemia-reperfusion injury in vivo,and the CCK-8 test was used to evaluate the protective effect of Safflower on H9c2cardiomyocytes after hypoxia-reoxygenation injury.The level of tumor necrosis factor TNF-αinflammation in cardiomyocyte medium was determined by ELISA.The effects of Safflower on the contents of lactate dehydrogenase（LDH）and superoxide dismutase（SOD）in cell medium were determined by biochemical kit.（3）Grey correlation analysis（GRA）and partial least square method（PLSR）were used to establish the"spectrum-effect"relationship between the Safflower fingerprint and biochemical indexes in vitro.Thirteen peaks of P11,P41,P40,P43,P23,P24,P26,P9,P45,P14,P13,P44,and P7 were positively correlated with the protective effect against myocardial ischemia-reperfusion injury.（4）Finally,the chromatographic peaks were identified by UPLC-Q-TOF-MS.The chromatographic peaks P7:Roseoside,P9:Quercetin-3,7-di-o-beta-d-Glucoside,P11:Hydroxysafflor yellow A,P13:Saffloquinoside C,P14:6-hydroxy kaempferol-3,6-di-o-beta-d-Glucoside,P23:Saffloquinoside A P24:Kaempferol-3-O-beta-sophorose,At the same time,other chromatographic peaks in the fingerprint were inferred the P3:Anthocyanidin,P6:Safflomin B,P15:Kaempferol-3-O-beta-rutinoside,P19:Quercetin-3-O-beta-rutinoside（Rutin）,P32:N¹,N⁵,N¹⁰-（E）-tri-p-coumaroylspermidine,P33:N¹,N⁵,N¹⁰-（Z）-tri-p-coumaroylspermidine.The chromatographic peaks P3,P23,and P24 have not been reported to be correlated with the protective effect of myocardischemia-reperfusionsion injury,which can be further verified in subsequent studies.Therefore,this study provides a reference for screening the potential active compounds of Safflower for protective effectsischemia-reperfusionsion injury.IIt providespossible data basis for clinical myocardial ischemia treatment and significance for subsequent quality evaluation of safflower compounds.