Inductive effect of hydroxyl safflower yellow-A on apoptosis in abnormal HUVEC via the mitochondrial pathway

Objective:To investigate the mechanism by which hydroxyl safflower yellow A,an active component of safflower (Carthamus tinctorius L.),promotes apoptosis in abnormal human umbilical vein endothelial cells (HUVECs).Methods:Supernatant of BGC-823 was used to stimulate HUVECs to establish a model of abnormal proliferation of HUVECs.After determining an ideal concentration of HSYA by MTT assay,apoptosis was detected with flow cytometry and TUNEL assay.Mechanism of apoptosis was assessed using quantitative real-time polymerase chain reaction,Western blot,and ELISA.Results:A range of concentrations of HSYA inhibited proliferation and promoted apoptosis of abnormal HUVECs.As the rate of apoptosis increased,mRNA expression of caspase-3 increased while expression of mutant p53 decreased.HSYA had no effect on Fas gene expression.Analogously,protein expression of Bax was increased while those of Bcl-2,Fas,and Fas-L were decreased.Conclusions:HSYA appears to induce apoptosis of HUVECs with the stimulation of the supematant of tumor cells.The mechanism of apoptosis by HSYA may involve activation of the mitochondrial apoptotic pathway and regulation of the expressions of Bcl-2,Bax,and p53.

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 Yellow Inhibits Progression of Hepatocellular Carcinoma by Modulating Immunological Tolerance via FAK Pathway

Objective To explore the anti-tumor effect of safflower yellow(SY)against hepatocellular carcinoma(HCC)and the underlying potential mechanism.Methods An in vitro model was established by mixing Luc-Hepa1-6 cells and CD3^(+)CD8^(+)T cells,followed by adding programmed cell death protein 1(PD-1)antibody(Anti-mPD-1)with or without SY.The apoptosis was detected by flow cytometry and the level of inflammatory cytokines was determined by enzyme-linked immunosorbent assay.The protein levels of programmed cell death 1 ligand 1(PD-L1),chemokine ligand(CCL5),C-X-C motif chemokine ligand 10(CXCL10)were measured by Western blot.An in situ animal model was established in mice followed by treatment with anti-mPD-1 with or without SY.Bioluminescence imaging was monitored with an AniView 100 imaging system.To establish the FAK-overexpressed Luc-Hepa1-6 cells,cells were transfected with adenovirus containing pcDNA3.1-FAK for 48 h.Results The fluorescence intensity,apoptotic rate,release of inflammatory cytokines,and CCL5/CXCL10 secretion were dramatically facilitated by anti-mPD-1(P<0.01),accompanied by an inactivation of PD-1/PD-L1 axis,which were extremely further enhanced by SY(P<0.05 or P<0.01).Increased fluorescence intensity,elevated percentage of CD3+CD8+T cells,facilitated release of inflammatory cytokines,inactivated PD-1/PD-L1 axis,and increased CCL5/CXCL10 secretion were observed in Anti-mPD-1 treated mice(P<0.01),which were markedly enhanced by SY(P<0.05 or P<0.01).Furthermore,the enhanced effects of SY on inhibiting tumor cell growth,facilitating apoptosis and inflammatory cytokine releasing,suppressing the PD-1/PD-L1 axis,and inducing the CCL5/CXCL10 secretion in Anti-mPD-1 treated mixture of Luc-Hepa1-6 cells and CD3+CD8+T cells were abolished by FAK overexpression(P<0.01).Conclusion SY inhibited the progression of HCC by mediating immunological tolerance through inhibiting FAK.

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.

Protective Effects of Hydroxysafflor Yellow A against Oxidative Damage of β-Mercaptoethanol During Neural Differentiation of Mesenchymal Stem Cells

Objective To study the protective effects of hydroxysafflor yellow A （HSYA） against the oxidative damage caused by β-mercaptoethanol （BME） during neural differentiation of mesenchymal stem cells （MSCs） in vitro. Methods When the confluence reached 50%-60%, 4th passage MSCs were divided into three groups to culture. Gt ： normal group which was cultured using basic medium （DMEM containing 10% FBS all the time）; G2： unprotected group which was continuously cultured using basic medium for 24 h, and then cultured using pre-induction medium （DMEM containing 10% FBS and 1 mmol/L BME）; G3： protected group which was firstly cultured using protective medium （DMEM containing 10% FBS and 160 mg/L HSYA） for 24 h, and then cultured using pre-induction medium for 24 h. After these treatments as above, cell viability, relative levels of SOD/GSH and apoptosis rate were respectively detected. The expression of Bcl and Bax was examined by Western blotting. After HSYA protection and BME pre-induction, neural induction was performed. The expression of NSE and MAP-2 was respectively analyzed on cellular and molecular levels. Results Compared with unprotected group, 160 mg/L HSYA could obviously improve cells viability, maintain high level of SOD and GSH in MSCs, reduce apoptosis rate and improve the ratio of Bcl/Bax. After protection with 160 mg/L HSYA, the survival time of neuron-like cells could be extended. Immunocytochemical staining showed that after 10 h of neural induction, the differentiated neuron-like cells in protected group were still in a good state, and the mRNA levels of NSE and MAP-2 were increased during the induction course checked. Conclusion HSYA could improve the resistance of cells to the oxidative damage caused by BME.