Effects of components in stasis-resolving and collateral-dredging Chinese herbal medicines on angiogenesis and inflammatory response of human umbilical vein endothelial cells induced by VEGF / 中国中药杂志

The present study investigated the mechanism of components in stasis-resolving and collateral-dredging Chinese herbal medicines, including scutellarin(Scu), paeonol(Pae), and hydroxy safflower yellow A(HSYA), in the treatment of psoriasis by regulating angiogenesis and inflammation. The human umbilical vein endothelial cells(HUVECs) cultured in vitro were divided into a normal group, a model group, a VEGFR tyrosine kinase inhibitor Ⅱ(VRI) group, and Scu, Pae, and HSYA groups with low, me-dium, and high doses. Cell viability was detected by the CCK-8 assay. Cell migration was detected by wound healing assay. Tube formation assay was used to measure the tube formation ability. Western blot was used to detect the protein expression of the VEGFR2/Akt/ERK1/2 signaling pathway. The secretion levels of inflammatory cytokines IFN-γ, IL-1β, IL-6, and TNF-α were detected by ELISA. The results showed that compared with the model group, all the Scu, Pae, and HSYA groups could reduce cell viability, inhibit cell migration and tube formation(P<0.05, P<0.01), and down-regulated the protein expression of VEGFR2, p-VEGFR2, Akt, p-Akt, ERK1/2, and p-ERK1/2. Scu and Pae could down-regulate VEGFR2 expression(P<0.05, P<0.01), while other groups only showed a downward trend. Scu and Pae significantly reduced IFN-γ and IL-6 levels(P<0.01), and HSYA significantly reduced the levels of IFN-γ, IL-1β, and IL-6(P<0.01). Scu, Pae, and HSYA had no significant effect on TNF-α. The results suggested that Scu, Pae, and HSYA may exert a therapeutic role in psoriasis-related angiogenesis and inflammation by inhibiting VEGFR2/Akt/ERK1/2 signaling pathway and inhibiting the secretion of IFN-γ, IL-1β, and IL-6.

Quantitative analysis of effects of salvianic acid a combined with hydroxy safflower yellow a on rat endothelial cells after hypoxic injury using the combination index method

Abstract Cerebrovascular disease is the second most serious disease in the world. It has the features of high morbidity, high mortality and recurrence rate. Numerous research on the compatibility of Chinese medicine with effective ingredients of cerebral ischemia has been made during the past decades. The purpose of this study is to quantitatively analyze the combined pharmacological effect of effective ingredients in Danshen and Honghua (Dan Hong) on rat microvascular endothelial cells after gradually oxygen-glucose deprivation. The experimental concentration range for the compatibility of two effective ingredients were determined in the preliminary experiments by Cell Counting kit-8 (CCK-8) method. Drugs were added to rat brain microvascular endothelial cells at a non-toxic dose level. After that, the cells were cultured for 12 h, and placed in a hypoxic environment. Finally, the cell survival rate was used as a measure of drug effect. In order to determine synergism or antagonism, the combination index (CI)-isobologram method was performed to analyze the data from the experiments. Based on this theory, the potencies of each drug and the shapes of their does-effect curves are both taken into account. The results show that the synergism or the antagonism between two effective ingredients compatibility change with different proportion and dosage. Furthermore, it can be seen from the results of these experiments that when these drugs are used in combination, the dosage required to achieve the same therapeutic effects is greatly reduced compared with the case of single one. It is worth mentioning that our experiments also prove that the median-effect equation and the CI method can be applied in the field of traditional Chinese medicine.

Preparation and quality evaluation of hydroxysafflor yellow A solid lipid nanoparticles / 国际中医中药杂志

Objective@#To optimize the formulation and preparation technology of hydroxy safflower yellow A solid lipid nanoparticles and evaluate its quality.@*Metheds@#Hydroxy safflower yellow A solid lipid nanoparticles were prepared by the method of hot melt emulsification ultrasonic-low temperature using hydroxypropyl methylcellulose as a lipid material and glyceryl monostearate as an emulsifier. Using entrapment efficiency as indexes, the amount of hydroxypropyl methylcellulose, purified water, glyceryl monostearate, and Tween 80 aqueous solution (1%) as factors, orthogonal test was applied to optimize the formulation and preparation technology. Dialysis method was used to measure encapsulation efficiency. The morphology and uniformity of the nanoparticles were observed by transmission scanning electron microscopy. The particle size, polydispersion index and Zeta potential were determined by nano-particle size analyzer. And hydroxy safflower yellow A solid lipid nanoparticles sustained releasing characteristics was evaluated by the percentage of cumulative drug release.@*Results@#The optimal process of prepared hydroxy safflower yellow A solid lipid nanoparticles was 5 mg of hydroxypropyl methylcellulose, 0.2 ml of purified water, 100 mg of glyceryl monostearate, and 1.8 ml of Tween 80 aqueous solution (1%). The size of the prepared nanoparticles was uniform and spherical. And the average particle size were (99.85 ± 3.04) nm, polydispersion index were (0.390 ± 0.021), Zeta potential were (-27.63 ± 2.12) mV, and the encapsulation efficiency of the hydroxy safflower yellow A solid lipid nanoparticles were (63.35 ± 2.65)%. The release rate of the nanoparticles was (44.35 ± 0.49)%.@*Conclusions@#The prepared hydroxy safflower yellow A solid lipid nanoparticles have good uniformity and good sustained release properties.

Preparation and quality evaluation of hydroxysafflor yellow A solid lipid nanoparticles / 国际中医中药杂志

Objective To optimize the formulation and preparation technology of hydroxy safflower yellow A solid lipid nanoparticles and evaluate its quality. Metheds Hydroxy safflower yellow A solid lipid nanoparticles were prepared by the method of hot melt emulsification ultrasonic-low temperature using hydroxypropyl methylcellulose as a lipid material and glyceryl monostearate as an emulsifier. Using entrapment efficiency as indexes, the amount of hydroxypropyl methylcellulose, purified water, glyceryl monostearate, and Tween 80 aqueous solution (1%) as factors, orthogonal test was applied to optimize the formulation and preparation technology. Dialysis method was used to measure encapsulation efficiency. The morphology and uniformity of the nanoparticles were observed by transmission scanning electron microscopy. The particle size, polydispersion index and Zeta potential were determined by nano-particle size analyzer. And hydroxy safflower yellow A solid lipid nanoparticles sustained releasing characteristics was evaluated by the percentage of cumulative drug release. Results The optimal process of prepared hydroxy safflower yellow A solid lipid nanoparticles was 5 mg of hydroxypropyl methylcellulose, 0.2 ml of purified water, 100 mg of glyceryl monostearate, and 1.8 ml of Tween 80 aqueous solution (1%). The size of the prepared nanoparticles was uniform and spherical. And the average particle size were (99.85 ± 3.04) nm, polydispersion index were (0.390 ± 0.021), Zeta potential were (-27.63 ± 2.12) mV, and the encapsulation efficiency of the hydroxy safflower yellow A solid lipid nanoparticles were (63.35 ± 2.65)%. The release rate of the nanoparticles was (44.35 ± 0.49)%. Conclusions The prepared hydroxy safflower yellow A solid lipid nanoparticles have good uniformity and good sustained release properties.

Protective Effects of Hydroxysafflor Yellow A against Oxidative Damage of β-Mercaptoethanol During Neural Differentiation of Mesenchymal Stem Cells / 中草药·英文版

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. G1: 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.

Adaptability research on ultrafiltration process of microfiltrated Qihong Maitong Injection / 中草药

Objective: To study the application of four kinds of hollow fiber ultrafiltration membranes by taking Qihong Maitong Injection (QMI) microfiltrated liquid as the research object. Methods: Polysulfone, polyether sulfone, polypropylene, and blended composite membranes with the entrapment relative molecular mass of 100000 for ultrafiltration were selected to determine the best appropriate ultrafiltration membrane material. The membrane flux of ultrafiltration was determined by taking the content of active ingredients (Astragalus total saponin, astragaloside IV, and hydroxy safflower yellow A), solid reduction rate, protein reduction rate, related substances, and pyrogen inspection of different membrane materials as the evaluation indexes. Results: The suitability of four different kinds of ultrafiltration membrane materials with the same entrapment relative molecular mass was different. The pure water flux recovery rates of polypropylene, polyether sulfone, and blended composite materials are higher than that of polysulfone material. The component permeation rates of polypropylene and polyether sulfone materials were higher, while the solid and protein reduction rates of polysulfone and blend compound materials were higher. For QMI, the ultrafiltration membrane with entrapment relative molecular mass of 100000 could effectively remove the pyrogen. Conclusion: The polypropylene-100000 ultrafiltration membrane could not only effectively remove the solid and high polymer material, but also keep the active ingredients. It is suitable for the purification of QMI.

The primary research on hydroxyl safflor yellow A injection mediating blood lipid / 国际中医中药杂志

Objective To investigate the effect of hydroxyl safflor yellow A on mediating blood lipid and to explore its primary mechanism.Methods Fifty KM mice were divided into five groups randomly:control group(A),hyperlipidemia model group(B),high-dose group(C),mid-dose group (D) and low-dose group(E).C,D and E group were injected by hydroxy safflor yellow A with 10,40 and 70 mg.kg-1 day-1respectively,while A and B group were both injected by saline with 0.4 ml day-1,the administrations were kept on three days.The levels of serum total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C) and malondialdehyde (MDA) were assayed,simultaneously the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were assayed after 18 hours.Results Compared with the control group,the serum TC (4.09+0.2) mmol/L,TG (0.96±0.15) mmol/L,LDL-C (5.87±0.17) mmol/L,HDL-C (0.83±0.21) mmol/L,MDA (8.26+1.05) nmol/ml,and the activities of SOD (330.18 ± 11.45 ) U/ml,GSH-Px (1023.54±25.34) U/ml of model group injected high doses of hydroxysafflor yellow A were statistically significant.Conclusion Hydroxy safflor yellow A had the function of mediating blood lipid.Anti-oxidation could be the mechanism.

Content Determination of Hydroxy Safflower Yellow A in Inner Mongolia Patent Medicine "Biyanqing"by HPLC / 国际中医中药杂志

Objective To establish content determination of hydroxyl safflower yellow A in Biyangqing.Methods Hhgh-performance liquid chromatography(HPLC)was used in the determination.A C18 column was used for the separation flow rate Was set at 1.0mL/min,the temperature of the column was set at 30℃,and wavelength of diction was set at 403 nm.with 100.08%average recovery and 0.98%RSD.Conclusion This detrmination method is specific and reproducible and can be used to control the quality of Biyangqing.

Pharmacokinetics of safflower yellow A in rats in vivo / 中草药

Object To study the pharmacokinetics of safflower yellow A, a major ingredient in the crude drug of Carthamus tinctorius L., in rats in vivo. Methods RP HPLC method was set up used to determine the content of safflower yellow A in plasma of rat. The chromatographic condition included Hypersil ODS2 column (200 mm?4.6 mm, 5 ?m); the mobile phase consisting of methanol 0.2% acetic acid (24∶76); the flow rate at 0.8 mL/min; the detective wavelength at 410 nm. Riboflavin was used as internal standard. Safflower yellow A was injected into the rats through caudal vein, and its concentrations at different time were determined in healthy rat after 24 h fasting. The data were processed with the software 3P87. Results The primary pharmacokinetic parameters were: V c (0 30?0 04) L/kg, CL (1.12?0.33) mL/min, K e (0.91?0.19) h -1 , t 1/2 (0 76?0 10) h, AUC (24.97?4.83) (?g?h)/mL, respectively in iv safflower yellow A to the rats. Conclusion The safflower yellow A in rats is fited to one compartment model, and it is distributed and eliminated quickly.

Thermolability in the process of safflower infusion solution by HPLC fingerprint / 中成药

AIM:To study incorporate changes of compound in safflower infusion solution before and after heating by HPLC fingerprint, in order to provide reference of drug’s safety evaluation. METHODS: Hydroxysafflor yellow A was used as reference substance, safflower infusion solution was detected by HPLC fingerprint in the whole manufacturing process from original raw material、intermediate to finished product.The detector temperature was set at room temperature、100 ℃、115 ℃,and then the change in the peak’ disappearance and appearance was observed and the peak area’ decrease and increase showed by characteristic absorption peak of every sample product at different temperature. RESULTS: As the temperature rose to 105 ℃,new peaks appeared at the retention time of 7、26-27、44 minutes,original peaks would disappear at the time 8、36minutes.As the temperature continued to rise to 115 ℃,the peak at the time of 7、10 minutes disappeared again.the peak area increased during the temperature rising at retention the time of 31-32、44 minutes and decreased at the retention time of 37 minutes. CONCLUSION: It indicates that,heating safflower infusion solution would produce a new compound. As the temperature continues to rise,the new compound would be destroyed.

Simultaneous determination of four kinds of components in Tongda Granule by HPLC-DAD / 中成药

AIM:To determine four bioactive components in Tongda Granule ( Radix Paeoniae alba,Flos Carthami,Radix Scutellariale,etc. ),namely hydroxy safflower yellow A,peoniflorin,ferulic acid,and baicalin. METHODS:The isolation was performed on an Alltech Apollo C18 (250 mm ? 4. 6 mm,5 ?m) with the mobile phase consisting of methanol-0. 2% (w)phosphoric acid for gradient elution. The flow rate was 1. 0 mL/min and the column temperature was set at 30 ℃. RESULTS:This method was successfully applied to determining four bio-active components in Tongda Granule. The standard curves were linear over the ranges of 5. 20 -104 mg/L for hydroxy safflower yellow A(r =0. 999 1),25. 6 -512 mg/L for peoniflorin (r =0. 999 6),7. 72 -154. 4 mg/L for ferulic acid (r =0. 999 5),32. 5 -650 mg/L for baicalin (r = 0. 999 5),respectively. The average recoveries were 101. 1% ,99. 6% ,98. 9% and 101. 9% ,RSD were 2. 3% ,1. 8% ,2. 6% and 1. 3% ,respectively. CONCLUSION:The method is simple and can be used as a quality control method for Tongda Granule.