Safflor yellow A protects vascular endothelial cells from ox-LDL-mediated damage.

Atherosclerosis is a chronic disease of arteries, which constitutes the pathological basis of a series of cardiovascular diseases. The inflammatory response of vascular endothelial cells mediated by oxidized low density lipoprotein (ox-LDL) is the early behavior and main signal of atherosclerosis. In this study, the damage model of vascular endothelial cells treated with ox-LDL was used to reproduce the damage process of vascular endothelial cells in the process of atherosclerosis. Cell viability was detected by CCK-8. The release levels of reactive oxygen species, nitric oxide, and superoxide dismutase (SOD) were detected by commercial kits. EdU cell proliferation assay was used to detect cell proliferation, real-time fluorescent quantitative PCR and Western blot were used to detect the expression level of related genes. The results showed we successfully constructed a vascular endothelial injury model by incubating vascular endothelial cells with gradient concentrations of ox-LDL. The incubation of safflor yellow A (SYA) partially restored the loss of viability of vascular endothelial cells mediated by ox-LDL, and SYA could promote the proliferation of injured vascular endothelial cells. In addition, SYA may transmit related signals through the AMPK pathway to protect vascular endothelial cells from ox-LDL-mediated damage. All these results provide a further understanding of the occurrence and development of atherosclerosis, provide a theoretical basis for the use of SYA-related drugs in the treatment of cardiovascular diseases, and provide a reference paradigm for studying the pharmacology, toxicology, and mechanism of action of key active substances in TCM.

[Corrigendum] Synergistic cardioprotective effects of Danshensu and hydroxysafflor yellow A against myocardial ischemia-reperfusion injury are mediated through the Akt/Nrf2/HO-1 pathway.

Subsequently to the publication of the above article, an interested reader drew to the authors' attention that some of the data panels shown in Fig. 6A on p. 90 appeared to contain overlapping data, such that the data may have been derived from the same original source where different experimental conditions were portrayed in the figure. The data that appeared to be overlapping were featured in the H/R and H/R+DH+Z panels (both the merged and the unmerged data panels). The authors have re-examined their original data and realize that they made an inadvertent error during the assembly of this figure. The corrected version of Fig. 6A, showing the correct TUNEL staining data for the H/R+DH+Z experiment, is shown on the next page. Note that the errors made during the assembly of this figure did not affect the major conclusions reported in the paper. All the authors have agreed to this Corrigendum, and thank the Editor of International Journal of Molecular Medicine for allowing them the opportunity to publish this. The authors regret these errors went unnoticed during the compilation of the figure in question, and apologize to the readership for any confusion that this may have caused. [the original article was published in International Journal of Molecular Medicine 38: 83-94, 2016; DOI: 10.3892/ijmm.2016.2584].

[Biological activity and hydroxy safflor yellow A content of intermediates in preparation of Safflower Injection].

In order to investigate the effect of various production processes on the quality of Safflower Injection, the biological activities of the intermediates were evaluated by measuring activated partial thromboplastin time (APTT) and adenosine diphosphate (ADP) induced platelet aggregation in vitro. Intermediates were produced by key processes, such as extraction, concentration, twice alcohol precipitation, water sedimentation and two sterilizations during the production of Safflower Injection. The content of main chemical components in intermediates was determined by HPLC. The results showed that with the advance of the preparation process of Safflower Injection, the inhibition of ADP-induced platelet aggregation rate of each intermediate decreased gradually, and the trend of extending APTT activity decreased first and then increased. Meanwhile, the content of hydroxy safflor yellow A (HSYA) was gradually lowered, the content of p-hydroxy-cinnamic acid was increased, and new chemical component p-hydroxybenzaldehyde was produced. In conclusion, sterilization played a key role in the biological activity and HSYA content of Safflower injection.

Biological activity and hydroxy safflor yellow A content of intermediates in preparation of Safflower Injection / 中国中药杂志

In order to investigate the effect of various production processes on the quality of Safflower Injection, the biological activities of the intermediates were evaluated by measuring activated partial thromboplastin time (APTT) and adenosine diphosphate (ADP) induced platelet aggregation in vitro. Intermediates were produced by key processes, such as extraction, concentration, twice alcohol precipitation, water sedimentation and two sterilizations during the production of Safflower Injection. The content of main chemical components in intermediates was determined by HPLC. The results showed that with the advance of the preparation process of Safflower Injection, the inhibition of ADP-induced platelet aggregation rate of each intermediate decreased gradually, and the trend of extending APTT activity decreased first and then increased. Meanwhile, the content of hydroxy safflor yellow A (HSYA) was gradually lowered, the content of p-hydroxy-cinnamic acid was increased, and new chemical component p-hydroxybenzaldehyde was produced. In conclusion, sterilization played a key role in the biological activity and HSYA content of Safflower injection.

Study on Improving Quality Standard of Qiwei Maqianzi Pills / 中国药房

OBJECTIVE:To improve the quality standard for Qiwei maqianzi pills.METHODS:TLC was used for the qualitative identification of Chebulae Fmctus and Aucklandiae Radix in the preparation.HPLC method was used for the content determination of hydroxy safflor yellow A,brucine and strychnine in preparation.The determination was performed on Phenomenex Prodigy C18 column with mobile phase consisted of methanol-acetonitrile-0.7％ phosphoric acid soulution(26 ∶ 2 ∶ 72,V/V/V,for hydroxy safflor yellow A),acetonitrile-0.01 mol/L sodium heptanesulfonate mixed with same quantity of 0.02 mol/L potassium dihydrogen phosphate (pH adjusted to 2.8 using 10％ phosphoric acid,21 ∶ 79,V/V,for brucine and strychnine) at the flow rate of 1.0 mL/min.The detection wavelengths were 403 nm (for hydroxy safflor yellow A) and 260 nm (for brucine and strychnine).The column temperature was 25 ℃C,and the injection volume was 10 μL.RESULTS:TLC spots of Chebulae Fructus and Aucklandiae Radix were clear and well-separated without interference from negative control.The linear range was 6.29-62.94 μg/mL for hydroxy safflor yellow A(r=0.999 3),1.83-18.30 μg/mL for brucine(r=0.999 4) and 2.11-21.11 μg/mL for strychnine (r=0.999 6).RSDs of precision,stability and reproducibility tests were lower than 2.0％.The recoveries were 101.66％-104.91％(RSD=1.14％,n=6),99.58％-104.55％ (RSD=1.75％,n=6) and 101.22％-104.04％ (RSD=0.99％,n=6),respectively.CONCLUSIONS:Improved standard can be better used for quality control of Qiwei maqianzi pills.

Quality Standard for Ehong Capsules / 中国药师

Objective: To establish the quality standard for Ehong capsules.Methods: A TLC was adopted to identify safflower, pueraria and prepared Radix Polygoni Multiflori.The contents of hydroxyl-safflor yellow A and puerarin in Ehong capsules were determined by a dual wavelength HPLC.A Wondasil C 18-WR (250 mm× 4.6 mm ,5 μm) was used as the analytical column, the mobile phase was composed of methanol acetonitrile-0.7% phosphoric acid aqueous solution (16∶3∶81), the flow rate was 1.0 ml·min-1 ,the detection wavelength was 403 nm for hydroxy safflower yellow A and 250 nm for puerarin.The column temperature was 30℃.Results: The TLC spots were clear and well-separated without negative interference.The calibration curves of hydroxyl-safflor yellow A and puerarin were linear over the ranges, the average recovery was 101.02% and 100.03% , and the RSD was 1.43% and 2.40% (n =6), respectively.Conclusion: The method is fast and accurate with good specificity, which can be used for the quality control of Ehong capsules.

Simultaneous determination of seven index components in Refined Coronary Tablet by HPLC / 中草药

Objective: To develop an HPLC method for the simultaneous determination of tanshinone IIA, paeoniflorin, salvianolic acid B, ferulic acid, safflor yellow A, ligustilide, and danshensu in Refined Coronary Tablets. Methods: The chromatographic separation was achieved on a Zorbax Eclipse XDB-C18 column (250 mm × 4.6 mm, 5.0 μm) with methanol-acetonitrile (25∶75, A)-0.1% formic acid (B) as mobile phases at the flow rate of 1.0 mL/min for gradient elution: 0-10.0 min, 95% B; 10.0-16.0 min, 95%-85% B; 16.0-18.0 min, 85% B; 18.0-22.0 min, 85%-75% B; 22.0-26.0 min, 75%-65% B; 26.0-40.0 min, 65%-15% B; Detection with variable wavelength: 0-19.0 min was 270 nm, 19.0-22.0 min was 230 nm, 22.0-27.0 min was 320 nm, 27.0-40.0 min was 402 nm, and the column temperature was 30 ℃. Its linear relationship, precision, repeatability, stability, and recoveries were investigated. Results: The results showed that the seven active components were well separated and showed good linearity, tanshinone IIA 0.4-8.0 mg/L (r = 0.999 5), paeoniflorin 1.2-24.0 mg/L (r = 0.999 1), salvianolic acid B 3.2-64.0 mg/L (r = 0.999 3), ferulic acid 0.08-1.60 mg/L (r = 0.999 5), safflor yellow A 1.2-24.0 mg/L (r = 0.999 3), ligustilide 0.24-4.80 mg/L (r = 0.999 7), and danshensu 0.32-6.40 mg/L (r = 0.999 7). The precision was good, and RSD was less than 2.0%. The repeatability was good, and RSD was less than 2.0%. The stability was good in 12 h. The average recoveries were between 98.05%-101.27%, and RSD was less than 2.0%. The contents of target components in Refined Coronary Tablets, tanshinone was 0.704-0.797 mg/g, paeoniflorin was 3.124-3.411 mg/g, salvianolic acid B was 7.129-7.611 mg/g, ferulic acid was 0.180-0.198 mg/g, safflor yellow A was 2.718-2.966 mg/g, ligustilide was 0.590-0.683 mg/g, and danshensu was 0.811-0.899 mg/g. Conclusion: The method is accurate, sensitive, credible, and repeatable. It can be applied to the quality control of Refined Coronary Tablets.

Hydroxy-safflor yellow A attenuates Aß1₋42-induced inflammation by modulating the JAK2/STAT3/NF-κB pathway.

Beta-amyloid (Aß)-mediated inflammation plays a critical role in the initiation and progression of Alzheimer׳s disease (AD). Anti-inflammatory treatment may provide therapeutic benefits. In this study, the effect of hydroxy-safflor yellow A (HSYA) on Aß1-42-induced inflammation in AD mice was investigated and the underlying mechanisms were explored. Aß1-42 was injected into bilateral hippocampi of mice to induce AD models in vivo. Spatial learning and memory of mice were investigated by the Morris water maze test. Activated microglia and astrocytes were examined by immunofluorescence staining for ionized calcium-binding adapter molecule-1 (Iba-1) and glial fibrillary acidic protein (GFAP). The mRNA of inflammatory cytokines were measured using real-time PCR. NF-κB p65 translocation was analyzed by western blotting and immunostaining. IκB and phosphorylation of JAK2 and STAT3 were tested by western blotting. The results showed that HSYA ameliorated the memory deficits in Aß1-42-induced AD mice. HSYA suppressed Aß1-42-induced activation of microglia and astrocytes and reduced the mRNA expression of pro-inflammatory mediators. HSYA up-regulated the JAK2/STAT3 pathway and inhibits the activation of NF-κB signaling pathways. Pharmacological inhibition of STAT3 by AG490 reversed the inactivation of p65 and anti-inflammatory effects of HSYA. In conclusion, these results suggest that HSYA protects Aß1-42-induced AD model through inhibiting inflammatory response, which may involve the JAK2/STAT3/NF-κB pathway.

Hydroxy-safflor yellow A inhibits neuroinflammation mediated by Aß1₋42 in BV-2 cells.

Inflammation is an important contributor to the development of Alzheimer's disease (AD). Anti-inflammatory medication may offer promising treatment for AD. Hydroxy-safflor yellow A (HSYA), a chemical component of the safflower yellow pigments, has been reported to exert potent immunosuppressive effects. This study examined the anti-inflammatory effects of HSYA in Aß1₋42-treated BV-2 microglia cells. The mRNA levels of IL-1ß, IL-4, IL-10, TNF-α, COX-2 and iNOS were detected by real-time PCR. Western blotting was used to determine the protein expression of COX-2, TNF-α, iNOS, Janus Kinase 2 (JAK2), p-JAK2, signal transducers and activators of transcription 3 (STAT3) and p-STAT3. BV2-conditioned medium was used to treat SH-SY5Y cells and primary neuronal cells in indirect toxicity experiments. Cell viability and apoptosis were assessed using MTT assay and Annexin V/PI staining respectively. The results demonstrated that HSYA significantly reduced the expression of the pro-inflammatory mediators and inhibited Aß1₋42-induced neuroinflammation. Moreover, HSYA protected primary cortical neurons and SH-SY5Y cells against microglia-mediated neurotoxicity. HSYA also enhanced the phosphorylation of JAK2/STAT3 pathway and inhibition of JAK2 by AG 490 attenuated the anti-inflammatory effects of HSYA. Overall, our findings suggested that HSYA inhibited Aß1₋42-induced inflammation and conferred neuroprotection partially through JAK2/STAT3 pathway, indicating that HSYA could be a potential drug for the treatment of AD.

Protective effect of intravenous infusion injection of safflor yellow and hydroxyl safflor yellow A on acute myocardial ischemia injury in rats / 国际中医中药杂志

Objective The comparison research of protective effect between intravenous infusion injection of safflor yellow and hydroxyl safflor yellow A on acute myocardial ischemia injury in rats. Methods Qualified 60 male Wistar rats were divided into 5 groups at random (12 in each group):Blank control group, AMI model (treated with normal saline) group, intravenous infusion injection of safflor yellow (90 mg/kg) group, HSYA low dosage (20 mg/kg) group and high dosage (40 mg/kg) group. The acute myocardial ischemia injury in rats was induced by ligating the anterior descending coronary artery in Wistar rats and administered different dosage of safflor yellow and hydroxyl safflor yellow A with intraperitoneal injection. Myocardial infarction degree (MID) was calculated by detecting myocardial infarction area with nitroblue tetrazolium(NBT) assay. The changes of ST-elevation, CK-MB, cTnT, SOD activities and MDA contents were detected and analyzed. Results The intravenous injection of safflor yellow and HSYA low dosage group can significantly decreased ST-elevation [difference is (0.087?.022)mv,(0.091?.014)mv],MID[difference is (20.13?.17)%,(18.36+9.38)%], CK-MB [difference is (1460.70+219.73)U/L, (1472.72?85.61)U/L], cTnT[difference is (2.345?.883)ng/ml, (2.358?.843)ng/ml], and MDA [difference is(5.71 ?.67) mmol/ml, (5.76?.84)mmol/ml] contents in serum, increased SOD[difference is(58.27?.99)U/ml,(56.49+5.19)U/ml]activities in serum.Conclusion It showed that intra venous injection of safflor yellow and hydroxyl safflor yellow A had the same protective effect on acute myocardial ischemia injury in rats. Hydroxyl safflor yellow A is an important portion of safflor yellow.

Vascular effect of hydroxyl-safflor yellow A and underlying mechanism / 上海第二医科大学学报

Objective To investigate the vascular effect of hydroxyl-safflor yellow A(HSYA) on rat thoracic aorta and its underlying mechanism. Methods The tension of isolated thoracic aorta rings of rats perfused with different concentrations of HSYA(1?10-6-1?10-4 mol/L) was measured using organ bath technique.The effects of HSYA on the vasocontraction induced by cumulative phenylephrine(PE)(1?10-6-1?10-4 mol/L),KCl(6?10-2 mol/L) and CaCl2(1?10-5-3?10-3 mol/L) were recorded respectively. Results HSYAcaused a concentration-dependent anti-contraction effects by KCl or PE in endothelium-intact and endothelium-denuded aortic rings.HSYA inhibited the CaCl2-induced contraction and downward shifted concentration-response curve of aortic rings.HSYA+HP resulted in more significant anti-contraction effect than single use of HSYA(P0.05).There were significant differences in anti-contraction effect between HSYA+RR and RR or HSYA(P

Determination of Hydroxy Safflor Yellow A in Orthopaedics Lotion 1 by RP-HPLC / 中国药房

OBJECTIVE:To establish a RP-HPLC method for the determination of hydroxy safflor yellow A in Orthopae_dics lotion 1.METHODS:The separation was performed on Phenomenex Luna C18 with mobile phase composed of methanol-acetonitrile-water(26∶ 2∶ 72).The flow rate was 1.0ml/min;the detective wavelength was 403nm and the column temperature was 30℃.RESULTS:At a sample size of 0.1 044? g～ 1.2 528? g(r=0.9 998),hydroxy safflor yellow A was noted to be of good linear relation with peak area score.The average recovery was 98.08%(RSD=0.52%).CONCLUSION:The method is simple,accurate,specific,sensitive,reproducible,and suitable for the quality control of Orthopaedics lotion 1.

Determination of the Content of Safflor Yellow A in Zhanjin Tinctura by RP-HPLC / 中药新药与临床药理

Objective To establish an RP-HPLC method for the determination of the content of safflor yellow A in Zhanjing Tinctura. Methods The chromatographic procedure was carried out in Symmetry C18 column(4.6mm?150 mm,5.0?m)with methanol-water-glacial acetic acid(7:93∶2)as the mobile phase,pH=2.80. The flow rate was 1.0 mL?min-1,column temperature was 20℃,and the detection wavelength was 402 nm. Results Safflor yellow A was well separated from other components. The standard curves of safflor yellow A showed linearity in the range of 0.096～0.960?g (r=0.9999). The average recovery was 98.80% with RSD being 2.95%(n=9). Conclusion This method is sensitive,simple and accurate,and can be used for the determination of safflor yellow A in Zhanjing Tinctura.

Correlation and path analysis on characters related to flower yield per plant of Carthamus tinctorius / 中草药

plant height.Multiple regression analysis showed that branch height from base(X_2),number of primary branches(X_4),number of effective cones per plant(X_6),number of ineffective cones per plant(X_7),number of grains per cone(X_9),and diameter of primary head(X_(10)) were the main factors affecting flower yield(Y) per plant.Multiple regression equation of flower yield per plant and six characters was Y=-3.037 0+(0.002 7) X_2+0.045 9 X_4+0.074 5 X_6+0.043 2 X_7+0.023 0 X_9+1.148 2 X_(10)(F=21.84~()).The direct effect of number of effective cones per plant was the strongest,followed by diameter of primary head.There were significant differences within the flower yield per plant and the safflor yellow A content of different species.The correlation between the safflor yellow A content and the flower yield per plant was insignificant.Conclusion High-yield and high-quality are compatible in breeding of safflower which is used as herbal medicine.Number of effective cones per plant and diameter of primary head are focused on the high yield breeding and cultivation of safflower species.The plant type of higher flower yield safflower species should have more effective cone numbers,more number of cones,number of branches,number of primary branches,bigger diameter of primary head,moderate plant height,branch height from base,and ineffective cones per plant.Of all accessions,PI 239226,PI 253540,PI 367833,and Jianyang Honghua are outstanding and optimal for cultivating in Sichuan Province.