3-4-dihydroxyphenyllactic-acid and protocatechuic-acid

Radix et Rhizoma Salviae Miltiorrhizae (Salvia miltiorrhiza Bge., Lamiaceae, Danshen in Chinese) and Chuanxiong Rhizoma (rhizomes of Ligusticum chuanxiong Hort., Apiaceae, Chuanxiong in Chinese) both are important traditional Chinese medicine (TCM) for activating blood and eliminating stasis. Danshen-chuanxiong herb pair has been used for more than 600 years in China. Guanxinning injection (GXN) is a Chinese clinical prescription refined from aqueous extract of Danshen and Chuanxiong at the ratio of 1:1 (w/w). GXN has been mainly used in the clinical therapy of angina, heart failure (HF) and chronic kidney disease in China for almost twenty years.. This study aimed to explore the role of GXN on renal fibrosis in heart failure mice and the regulation of GXN on SLC7A11/GPX4 axis.. The transverse aortic constriction model was used to mimic HF accompanied by kidney fibrosis model. GXN was administrated by tail vein injection in dose of 12.0, 6.0, 3.0 mL/kg, respectively. Telmisartan (6.1 mg/kg, gavage) was used as a positive control drug. Cardiac ultrasound indexes of ejection fraction (EF), cardiac output (CO), left ventricle volume (LV Vol), HF biomarker of pro-B type natriuretic peptide (Pro-BNP), kidney function index of serum creatinine (Scr), kidney fibrosis index of collagen volume fraction (CVF) and connective tissue growth factor (CTGF) were evaluated and contrasted. Metabolomic method was employed to analyze the endogenous metabolites changes in kidneys. Besides, contents of catalase (CAT), xanthine oxidase (XOD), nitricoxidesynthase (NOS), glutathione peroxidase 4 (GPX4), the x(c)(-) cysteine/glutamate antiporter (SLC7A11) and ferritin heavy chain (FTH1) in kidney were quantitatively analyzed. In addition, ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to analyze the chemical composition of GXN and network pharmacology was used to predict possible mechanisms and the active ingredients of GXN.. The cardiac function indexes of EF, CO and LV Vol, kidney functional indicators of Scr, the degree of kidney fibrosis indicators CVF and CTGF were all relieved to different extent for the model mice treated with GXN. 21 differential metabolites involved in redox regulation, energy metabolism, organic acid metabolism, nucleotide metabolism, etc were identified. Aspartic acid, homocysteine, glycine, and serine, methionine, purine, phenylalanine and tyrosine metabolism were found to be the core redox metabolic pathways regulated by GXN. Furthermore, GXN were found to increase CAT content, upregulate GPX4, SLC7A11 and FTH1 expression in kidney significantly. Not only that, GXN also showed good effect in down-regulating XOD and NOS contents in kidney. Besides, 35 chemical constituents were initially identified in GXN. Active ingredients of GXN-targets-related enzymes/transporters-metabolites network was established to find out that GPX4 was a core protein for GXN and the top 10 active ingredients with the most relevant to renal protective effects of GXN were rosmarinic acid, caffeic acid, ferulic acid, senkyunolide E, protocatechualdehyde, protocatechuic acid, danshensu, L-Ile, vanillic acid, salvianolic acid A.. GXN could significantly maintain cardiac function and alleviate the progression of fibrosis in the kidney for HF mice, and the mechanisms of action were related to regulating redox metabolism of aspartate, glycine, serine, and cystine metabolism and SLC7A11/GPX4 axis in kidney. The cardio-renal protective effect of GXN may be attributed to multi-components like rosmarinic acid, caffeic acid, ferulic acid, senkyunolide E, protocatechualdehyde, protocatechuic acid, danshensu, L-Ile, vanillic acid, salvianolic acid A et al.

Topics: Animals; Chromatography, Liquid; Drugs, Chinese Herbal; Fibrosis; Glycine; Heart Failure; Mice; Rosmarinic Acid; Salvia miltiorrhiza; Tandem Mass Spectrometry; Vanillic Acid

A simple, convenient, and sensitive method that involves combining matrix solid-phase dispersion extraction with field-amplified sample stacking in capillary electrophoresis has been developed for the determination of organic acids in a complex solid matrix. Mesoporous molecular sieve, MCM-48, was synthesized by a hydrothermal method and selected as the adsorbent in matrix solid-phase dispersion. After fast extraction, the enriched analytes were back-extracted into a basic aqueous solution for field-amplified sample stacking in capillary electrophoresis. Parameters that affect extraction efficiency and sample stacking were optimized. Under the optimal conditions, approximately 42-, 49-, and 56-fold sensitivity enhancements were achieved for danshensu, protocatechuic acid, and cinnamic acid, respectively, when compared to normal injection. A satisfactory correlation coefficient (r > 0.99) was obtained. Both intra- and interday precision were lower than 2.53%. And the limits of detection of the three organic acids ranged between 0.01 and 0.029 μg/mL. Finally, the newly proposed method was successfully applied to the determination of organic acids in Fufang Danshen tablets, which indicates its great potential in analyzing organic acids in a complex matrix.

Topics: Adsorption; Cinnamates; Electrophoresis, Capillary; Hydroxybenzoates; Lactates; Particle Size; Porosity; Solid Phase Extraction; Surface Properties

Topics: Administration, Oral; Benzaldehydes; Camphanes; Catechols; China; Drugs, Chinese Herbal; Humans; Hydroxybenzoates; Lactates; Lactic Acid; Panax notoginseng; Salvia miltiorrhiza; Tandem Mass Spectrometry

To explore potency material bases of Xuebijing (XBJ) formula, and to analyze its effects at the molecular network level.. Totally 16 sepsis-related targets were selected and classified into three categories such as inflammation, immune, and coagulation referring to biological roles. Then molecular database of chemical compositions in XBJ formula were constructed to explore mutual actions with inflammation, immune, and coagulation targets.. Danshen root and safflower, with more effector molecules with immune and coagulation targets, have extensive anticoagulation and anti-inflammation effects. The former 10 molecules with better mutual actions with sepsis targets were sequenced as tryptophane, danshensu, gallic acid, salvianolic acid D, protocatechuic acid, salvianolic acid A, danshensu C, vanillic acid, rosmarinic acid, phenylalanine. There existed two phenomena in XBJ formula as follows. One component had stronger actions with multi-targets, for example, danshensu had actions with 13 targets. Meanwhile, different components acted on the same target protein, for example, 8 molecules acted with MD-2.. XBJ formula had certain potential synergistic effects with sepsis targets, which could provide certain referential roles for findina new type anti-septic drugs.

Topics: Caffeic Acids; Drugs, Chinese Herbal; Gallic Acid; Hydroxybenzoates; Inflammation; Lactates; Sepsis

To evaluate the pharmacokinetics of phenolic compounds after oral administration of Danshen extract in rat brain.. Blood and brain microdialysis probes were inserted into jugular vein and cerebral cortex of rat under anesthesia and perfused with ringer's solution at the rate of 2.0 and 0.8 μL/min, respectively. Blank microdialysates were collected after 2h post-implantation equilibrium time. Danshen extract (danshensu 40 mg/kg BW, protocatechuic aldehyde 149 mg/kg BW, and salvianolic acid B 50mg/kg BW) was administrated intragastrically, and then blood and brain microdialysates were collected at 15 and 30 min time intervals for 4h, respectively. The concentrations of phenolic compounds were determined by high-performance liquid chromatography coupled with chemiluminescence detection. Pharmacokinetic parameters were estimated using non-compartmental methods.. Danshensu and protocatechuic acid could be detected in both blood and brain microdialysates, while protocatechuic aldehyde and salvianolic acid B were not detected. Brain-to-blood (AUC(brain)/AUC(blood)) distribution ratio were 0.25±0.04 and 0.09±0.02 for danshensu and protocatechuic acid, respectively.. Danshensu can readily permeate the blood brain barrier after oral administration of Danshen extract, and protocatechuic acid is a potential oxidative metabolite of protocatechuic aldehyde.

Topics: Animals; Benzaldehydes; Benzofurans; Blood-Brain Barrier; Catechols; Drugs, Chinese Herbal; Hydroxybenzoates; Lactates; Male; Microdialysis; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza

A simple and sensitive capillary electrophoresis method with field-enhanced stacking concentration for the analysis of protocatechuic aldehyde, protocatechuic acid, danshensu, rosmarinic acid and salvianolic acid B in Salvia miltiorrhiza var. miltiorrhiza f. alba was developed. The separation was achieved with a fused-silica capillary (75 microm x 50.2 cm, effective length was 40 cm) and a running buffer 15 mmol x L(-1) borax (pH 10.0) containing 20% CH3 OH. The UV detection wavelength was 210 nm. The applied voltage was 28 kV, and the cartridge temperature was 25 degrees C. Water plug was introduced from the anode by 0.5 psi x 4 s before injection. Sample was injected by electrokinetic injection - 8 kV x 3 s. The linear range of protocatechuic aldehyde is 3.0-60.00 mg x L(-1) (R2 = 0.999 8); that of protocatechuic acid, danshensu, rosmarinic acid and salvianolic acid B are 1.0-20.00 mg x L(-1) (R2 are 0.999 1, 0.999 4, 0.998 9 and 0.999 8, respectively), and the limits of detection of five analyts are 0.55, 0.40, 0.25, 0.32, 0.38 microg x L(-1), respectively, Stacking factor is higher and precision is satisfactory. The recoveries ranges were from 97.3% to 99.8%. The proposed method was used to determine the protocatechuic aldehyde, protocatechuic acid, danshensu, rosmarinic acid and salvianolic acid B in S. miltiorrhiza var. miltiorrhiza f. alba. The proposed method is simple, rapid, accurate and high sensitivity, and can be used to control of the quality of S. miltiorrhiza var. miltiorrhiza f. alba.

Topics: Benzaldehydes; Benzofurans; Catechols; Cinnamates; Depsides; Electrophoresis, Capillary; Hydroxybenzoates; Lactates; Plant Extracts; Quality Control; Rosmarinic Acid; Salvia miltiorrhiza; Sensitivity and Specificity; Solubility; Water

We have used microarray technology to detect the effect of Guanxin No.2 decoction on gene expression in different areas of the myocardial infarcted heart of rats.. Male Sprague-Dawley rats (180-200 g) were randomly divided into three groups: sham-operated; coronary artery ligation; and coronary artery ligation plus administration of Guanxin No.2 decoction (10.0 g raw materials/kg per day by gavage). The experiment was carried out on day seven after ligation.. We found that the gene expression using microarray technology showed many differences in the border infarcted left ventricular area compared with the remote noninfarcted left ventricular area after administration of Guanxin No.2 decoction.. Guanxin No.2 decoction has a long history in treating ischaemic cardiomyopathy in China, but the molecular mechanism has been unclear. In this study we found that some important genes may have contributed to the cardioprotective effect of Guanxin No.2 decoction.

Topics: Animals; Benzaldehydes; Benzoates; Benzofurans; Bridged-Ring Compounds; Cardiotonic Agents; Carthamus tinctorius; Catechols; Chalcone; Coumaric Acids; Dalbergia; Drugs, Chinese Herbal; Gene Expression; Gene Expression Profiling; Glucosides; Heart Ventricles; Hydroxybenzoates; Lactates; Ligusticum; Male; Monoterpenes; Myocardial Infarction; Oligonucleotide Array Sequence Analysis; Paeonia; Plant Extracts; Quinones; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; Salvia miltiorrhiza

To establish a high performance capillary zone electrophoretic method for the separation of water-soluble active components, protocatechuic aldehyde (PAH), danshensu (DSS), and protocatechuic acid (PA), in Salvia miltiorrhiza bge. f. alba, many factors that can affect the resolution and the migration time, including the concentration of phosphate-borax buffer, pH value, separation voltage and detection wavelength were investigated. The optimized conditions were as follows: uncoated capillary column, 75 microm x 60.2 cm (effective length of 50 cm); column temperature, 25 degrees C; detection wavelength of the photo-diode-array detector, 210 nm; injection, 3.45 kPa x 4 s; separation voltage, 27.5 kV; running buffer, 5 mmol/L borax (pH 7.5). Under the optimized conditions, good separation of PAH, DSS and PA can be achieved in less than 8 min.

Topics: Benzaldehydes; Catechols; Electrophoresis, Capillary; Hydrogen-Ion Concentration; Hydroxybenzoates; Lactates; Salvia miltiorrhiza; Solubility; Water

Topics: Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Drugs, Chinese Herbal; Hydroxybenzoates; Lactates

Homocysteine (Hcy) is a by-product of methionine metabolism. An imbalance of Hcy in the body may lead to hyperhomocysteinemia, a condition with elevated Hcy concentration in blood that may be one of the risk factors responsible for the development of several vascular diseases (thromboembolism, atherosclerosis, stroke, vascular diseases and dementia). Radix Salvia miltiorrhiza (Danshen), a well-known Chinese medicinal herb that can activate and improve blood microcirculation, is noticeable for its beneficial effect in treating cardiovascular diseases. The present study is to demonstrate the protective effect of Danshen extract against the homocysteine-induced adverse effect on human umbilical vein endothelial cell (HUVEC). Homocysteine (5 mM) not only decreased the cell viability but also caused the disruption of capillary-like structure formation in vitro. The protective effect of Danshen aqueous extract and its active compounds on endothelial cell function were demonstrated through an in vitro tube formation assay, which mimics the new blood vessel formation. To identify the active components in the aqueous extract of Danshen, the content was characterized by instrumental analysis using high performance liquid chromatography with diode array detector (DAD) and electrospray tandem mass spectrometry (ESI-MS/MS). Interestingly, Danshen extract and its pure compounds showed different effectiveness in protecting HUVEC against Hcy-induced injury according to the following descending order: Danshen aqueous extract, 3-(3,4-dihydroxy-phenyl)-2-hydroxy-propionic acid (Danshensu), protocatechuic acid, catechin and protocatechualdehyde. We believed that such findings might provide evidence in understanding the beneficial effects of Danshen on the cardiovascular system.

Topics: Benzaldehydes; Catechin; Catechols; Cell Survival; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Endothelial Cells; Homocysteine; Humans; Hydroxybenzoates; Lactates; Salvia miltiorrhiza; Spectrometry, Mass, Electrospray Ionization; Umbilical Veins

The effects of seven phenolic compounds isolated from Salvia miltiorrhiza on peroxidative damage to liver microsomes, hepatocytes and erythrocytes of rats were studied. The results show that the seven compounds inhibited lipid peroxidation of rat liver microsomes induced by iron/cysteine and Vitamin C/NADPH. The hemolysis of rat erythrocytes induced by hydrogen peroxide was also inhibited. The degree of inhibition varied with different compounds. Among the seven compounds, the action of salvianolic acid A (Sai A) was the most potent. Therefore, the protective action of Sai A against peroxidative damage to isolated rat hepatocytes and their plasma membranes was evaluated further. Malondialdehyde (MDA) production and bleb of the surfaces of rat hepatocytes induced by iron/cysteine were prevented by Sai A. The production of MDA and the consumption of NADPH of the plasma membrane during lipid peroxidation initiated by iron/cysteine and Vitamin C/NADPH were also inhibited. The results strongly suggest that several phenolic compounds like Sai A have a protective action against peroxidative damage to biomembranes.

Topics: Animals; Benzaldehydes; Caffeic Acids; Catechols; Cinnamates; Depsides; Erythrocyte Membrane; Hydroxybenzoates; Lactates; Lipid Peroxidation; Liver; Male; Microscopy, Electron, Scanning; Microsomes, Liver; Rats; Rats, Inbred Strains; Rosmarinic Acid

Topics: Anticoagulants; Benzaldehydes; Catechols; Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Hydroxybenzoates; Injections; Lactates; Phenanthrolines; Plant Extracts; Salvia miltiorrhiza