Stability and degradation of hydroxysafflor yellow A and anhydrosafflor yellow B in the Safflower injection studied by HPLC-DAD-ESI-MS^n

Safflower is a popular Chinese medicinal plant and Safflower injection is extensively used for the clinical treatment of cerebrovascular and cardiovascular diseases. In this study, HPLC-DAD-ESI-MSn was utilized to study the stability and degradation of the two major but chemically unstable bioactive compounds hydroxysaffior yellow A and anhydrosaffior yellow B, in Safflower injection. The impact of light irradiation, temperature, and pH on the stability of these two compounds were studied. The results showed that hydroxysafflor yellow A and anhydrosafflor yellow B could degrade at high temperature （〉60 ℃） or extreme pHs （pH ≤ 3.0 or 〉7.0）, but not under light irradiation. The common degradation product was p-coumaric acid. Chemical structures of the other degradation products were characterized by LC-MS. Hypothetical degradation pathways were proposed. In addition, ADP-induced platelet aggregation tests showed that the degradation of anhydrosaffior yellow B could reduce the anticoagulation activities of Safflower injection. Our results suggest that temperature and pH are critically important for the preparation and storage of Safflower injection.

Simultaneous determination of four nucleosides in Carthamus tinctorius L.and Safflower injection using high-performance liquid chromatography

We quantitatively determined four nucleosides, including cytidine, uridine, guanosine, and adenosine, in Carthamus tinctorius L. and Safflower injection. Separation was performed on a Zorbax Eclipse XDB-18 column using a gradient elution with mobile phases of 0.05% trifluoroacetic acid （TFA） aqueous solution （A） and methanol （B）. The assay was carried out at a flow rate of 1 mL/min at 25 ℃ with detection at 260 nm. Cytidine, uridine, adenosine and guanosine showed good linearity in the ranges of4.02-503μg/mL （r2= 0.9998）, 9.38-1407 μg/mL （rz = 0.9999）, 80.6-8060μg/mL （r2 = 0.9999） and 2.10---630μg/mL （r2 = 0.9987） with average recoveries of 97.2%, 94.5%, 98.6% and 108.6%, respectively. The contents of cytidine, uridine, adenosine and guanosine in different Carthamus tinctorius L. and Safflower injection were significantly different. This is the first report on the quantitative determination of nucleosides in Carthamus tinctorius L. and Safflower injection.

Screening Safflower Injection for Constituents with Activity against Stroke Using Comprehensive Chemical Profiling Coupled with Network Pharmacology

Objective:This study aimed to explore safflower injection(SI)for constituents with activity against ischemic stroke using a combination of chemical analysis and a network pharmacology strategy.Materials and Methods:The main ingredients of SI were comprehensively identified using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry,and the core targets and pathways associated with stroke were predicted using PharmMapper and Kyoto Encyclopedia of Genes and Genomes analysis.Cytoscape software was used to visualize and analyze the active compound-target-pathway network of SI regulating ischemic stroke.Results:A total of76 chemical compounds were identified from the SI sample,including 63,which regulated 88 targets that were ultimately enriched in 12 key ischemia stroke-related signaling pathways.Kaempferol-3-O-sophoroside,kaempferol-3-O-rutinoside,carthamoside B6,neoeriocitrin,and6-hydroxykaempferol-3-O-rutinoside-6-O-glucoside were determined to be important for stroke treatment because they had a higher degree value in the network than other constituents did.Moreover,the characteristic components isolated from SI showed protective effect mainly by acting on multiple targets including AKT1,epidermal growth factor receptor,transforming growth factor-beta receptor(TGFBR),Ras homolog,mTORC1 binding,caspase 3,and glycogen synthase kinase 3 beta,which were involved in different signaling pathways including phosphoinositide 3-kinase-Akt,mitogen-activated protein kinase,neurotrophin,ErbB,mechanistic target of rapamycin,and tumor necrosis factor.Conclusions:This study proposed a network pharmacology and chemical component profiling strategy for the systematic understanding of the therapeutic material basis of using SI against ischemic stroke.

A Clinical Study of Safflower Yellow Injection in Treating Coronary Heart Disease Angina Pectoris with Xin-Blood Stagnation Syndrome

Objective： To evaluate the clinical effect and safety of Safflower Yellow injection （SYI） in treating coronary heart disease angina pectoris （OHD-AP） with Xin-blood stagnation syndrome （XBSS）. Methods： Adopted was the multi-centered, randomized, positive parallel controlled method, 448 patients with CHD-AP-XBSS were enrolled and divided into two groups, 336 in the tested group treated with SYI and 112 in the control group treated with Salvia injection by intravenous dripping once a day for 14 days, so as to observe the conditions of angina, electrocardiogram, and therapeutic effect on traditinal Chinese medicine （TCM） symptoms as well as the safety of the treatment. Results： The significantly effective rate and total effective rate in the tested group were 60.06% （194/323） and 91.02 % （294/323） respectively; those in improvement of TOM symptoms were 40. 18% （129/321） and 75.23% （243/323） respectively, which were better than those in the control group （P〈0.01）. Conclusion： SYI Injection is effective and safe in treating OHD-AP-XBSS.

Safflower Extract and Aceglutamide Injection Promoting Recovery of Peripheral Innervations via Vascular Endothelial Growth Factor-B Signaling in Diabetic Mice

Background： Safflower extract and aceglutamide （SA） has been used clinically for the treatment of cerebrovascular diseases such as cerebral embolism, hemorrhage, and mental deterioration. This study aimed to investigate the effect and mechanism of SA injection in the recovery of peripheral innervations of diabetic mice. Methods： The C57BL/6 male mice were divided into four groups： normal control group （n = 44）, diabetic group （n = 44）, diabetic ＋ SA group （diabetic mice treated with SA injection, n = 44）, and diabetic ＋ SA ＋ vascular endothelial growth factor receptor （VEGFR）1-BL group （diabetic mice treated with SA injection and VEGFR 1 blocking antibody n = 24）. The streptozotocin-induced diabetic mice model and injured peripheral nerve mice model were built. The mice with injured peripheral nerves were intraperitonealy administered with SA injection for successive 21 days. The corneal sensitivity, number of corneal nerve fibers, and contents of vascular endothelial growth factor （VEGF）-B and various neurotrophic factors such as nerve growth factor （NGF） and glial cell line-derived neurotrophic factor （GDNF） in corneal tissue of four groups were observed. Results： The diabetic group showed decreased number of corneal nerve fibers, compared with the control group （P = 0.002）. And compared with the diabetic group, the diabetic ＋ SA group showed a significant increase in the number of nerve fibers （P = 0.024） and the contents of VEGF-B,NGF,andGDNFinthecornea（allP〈0.05）.However,whenthediabeticmiceweretreatedwiththeblockingantibodiesspecializedfor VEGF-B receptor, the neutralization ofVEGFR-1 completely abolished the increased expression of NGF and GDNF stimulated by SAinjection. Conclusions： SA injection could reduce the nerve injury caused by diabetic peripheral neuropathy, and its protective effect might be associated with the promotion of the expressions of VEGF-B, NGF, and GDNF.

Safflower yellow injection combined with conventional therapy in treating unstable angina pectoris:a meta-analysis

OBJECTIVE： To evaluate the clinical efficacy of safflower yellow injection combined with conventional therapy in treating unstable angina pectoris.METHODS： We searched online databases： Chinese journal full-text database, China National Knowledge Infrastructure, Wanfang database, Chinese journal full-text database, Pubmed, ScienceDirect,Embase, and the Cochrane Library with manual-screening of relevant literature. Eligible randomized controlled trials（RCT） on angina pectoris were included. We conducted meta-analysis using the RevMan 5.1 software from The Cochrane Collaboration. We treated the relief rate of angina symptoms and electrocardiograph（ECG） as evaluation.RESULTS： Seven articles, including in 1134 patients, were enrolled after the evaluation. Therewas no significant heterogeneity among the studies（χ2=1.08, df=6, P=0.98, I2=0%）. The safflower yellow injection with conventional therapy has a higher effective rate than the control group in relieving the symptoms of angina pectoris [odds ratio（OR）=2.95, 95%（CI）（1.81, 4.81）] and improving ischemic ECG [OR=2.85, 95% CI（1.67, 4.86）]. The difference was statistically significant in the ＂80 mg dosage＂ and ＂100 mg dosage＂ subgroups（P0.05） for improving clinical symptoms and ECG. The funnel graphic was nearly symmetrical. Sensitivity analysis suggested that the results were stable.CONCLUSION： Safflower yellow injection as an adjunct therapy with conventional drugs shows advantages in easing the clinical symptoms of unstable angina and improving ECG over basic therapy alone. However, the conclusions should be interpreted with care until more high-quality RCTs are reported.