salvianolic-acid-a and protocatechualdehyde

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

Topics: Benzaldehydes; Benzofurans; Bilirubin; Caffeic Acids; Catechols; Cinnamates; Depsides; Glucuronosyltransferase; Humans; Kinetics; Lactates; Microsomes, Liver; Polyphenols; Rosmarinic Acid; Salvia miltiorrhiza

Topics: Alkenes; Animals; Benzaldehydes; Caffeic Acids; Catechols; Chromatography, High Pressure Liquid; Cinnamates; Depsides; Drugs, Chinese Herbal; Ginsenosides; Glucosides; Hydroxybenzoates; Isoflavones; Limit of Detection; Male; Polyphenols; Rats; Rats, Sprague-Dawley; Rosmarinic Acid; Saponins; Tandem Mass Spectrometry; Triterpenes

Salvianolic acids, the well-known active components in Salvia miltiorrhiza, have been shown to possess markedly pharmacological activities. However, due to the complex in vivo course after administration, the pharmacologically active forms are still poorly understood. In present study, we evaluated the stability of eight major salvianolic acids from Danshen extract under different chemical and physiological conditions. We also quantitatively explained the absorption, metabolism and excretion of these salvianolic acids in rats after gastric-administration, which was carried out by simultaneously determining the amounts of salvianolic acids and their metabolites in the rat gastrointestinal contents, gastrointestinal mucosa, plasma, bile and urine. We found that: 1) protocatechuic aldehyde (PAL) was much stable whether in acidic environment (pH4.0) or in alkaline environment (pH8.0), while other salvianolic acids were stable in acidic environment and instable in alkaline environment; 2) PAL, salvianoli acid A (SAA) and salvianolic acid B (SAB) were instable whether in rat stomach or in small intestine, while other salvianolic acids were stable in rat stomach and instable in small intestine; 3) after gastric-administration, except PAL and Danshensu (DSS), other phenolic acids would be metabolized into DSS and caffeic acid (CA) in the rat gastrointestinal tract before absorption, and only free and glucuronidated PAL, CA and DSS were detected in rat plasma, bile and urine. In conclusion, it was the free and glucuronidated PAL, CA and DSS rather than the prototypes of other salvianolic acids that were present in plasma with considerable concentrations after gastric-administration.

Topics: Alkenes; Animals; Benzaldehydes; Caffeic Acids; Catechols; Drug Stability; Drugs, Chinese Herbal; Hydroxybenzoates; Lactates; Male; Metabolome; Molecular Structure; Polyphenols; Rats; Rats, Wistar; Salvia miltiorrhiza

An ultra high performance liquid chromatography with photodiode array detection method is developed for the simultaneous quantitative determination of five water-soluble compounds including danshensu, protocatechualdehyde, rosmarinic acid, salvianolic acid B, and salvianolic acid A in Salvia miltiorrhiza Bge.. Through method optimization, the five compounds all expressed good linearity (R(2) > 0.9990) in a wide concentration range together with satisfactory accuracy, precision, and stability. Moreover, through qualitative analysis of the chemical fingerprint combined with similarity analysis, hierarchical cluster analysis, principle component analysis, and partial least-squares discriminate analysis, we determined that the 13 batches of Salvia miltiorrhiza Bge. were similar in internal quality and the differences resulted from various cultivation environments, recovery elements, and others. Seen from the results of hierarchical cluster analysis and principle component analysis, the classification of 13 batches was in accordance, and partial least-squares discriminate analysis technique was more suitable than the principle component analysis model to provide a distinct classification of test samples on the basis of their different components. Moreover, a permutation test verified the rationality of partial least-squares discriminate analysis and variable importance plot showed that peaks 37 and 38 were the most significant variables in distinguishing the Salvia miltiorrhiza Bge.. The idea of the quantitative and qualitative analysis of Salvia miltiorrhiza Bge. was convenient, sensitive, and comprehensive, which could be applied to evaluate the quality of more traditional Chinese medicines.

Topics: Benzaldehydes; Benzofurans; Caffeic Acids; Catechols; Chromatography, High Pressure Liquid; Cinnamates; Depsides; Lactates; Least-Squares Analysis; Principal Component Analysis; Rosmarinic Acid; Salvia miltiorrhiza

To develop a LC-MS/MS method for the determination of protocatechuic acid, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone and tanshinone II(A) in rat plasma and brain. The plasma and brain samples were precipitated with ethyl acetate, then were separated on an Agilent eclipse plus-C18 column (2.1 mm x 50 mm, 3.5 microm) using acetonitrile (consisting of 0.1% formic acid) and water (consisting of 0.1% formic acid) as mobile phase in gradient elution mode. The mass spectrometer was operated under both positive and negative ion mode with the ESI source, and the detection was performed by MRM. The transition of 154.3/153.1 m/z for protocatechuic acid, 137.3/108 m/z for protocatechuic aldehyde, 493.0/295.2 m/z for Salvianolic acid A, 718.0/520.0 m/z for salvianolic acid B, 321.4/152.3 m/z for chloramphenicol, 297.4/254.3 m/z for cryptotanshinone, 295.5/249.3 m/z for tanshinone II(A) and 285.2/154.0 m/z for Diazepam. The calibration curves in the range of 0.625-1 000 microg x L(-1) for protocatechuic acid and protocatechuic aldehyde, 1.25-1 000 microg x L(-1) for salvianolic acid A, 2.5-1 000 microg x L(-1) for salvianolic acid B, 0.15-1 000 microg x L(-1) for cryptotanshinone, 0.625-1 000 microg x L(-1) for tanshinone II(A) are with good linearityin rat plasma and brain. The analysis method is sensitive, simple, and suitable enough to be applied in the pharmacokinetic study of the 6 main components. Animal testing gives the lgBB of the drugs and further studies of the 6 components cross the blood-brain barrier can be carried out.

Topics: Abietanes; Animals; Benzaldehydes; Benzofurans; Blood-Brain Barrier; Brain; Caffeic Acids; Catechols; Chromatography, Liquid; Hydroxybenzoates; Injections, Intravenous; Lactates; Phenanthrenes; Plant Preparations; Rats; Reproducibility of Results; Salvia miltiorrhiza; Tandem Mass Spectrometry

Salvianolic acid A, salvianolic acid B, danshensu, protocatechuic aldehyde, rosmarinic acid and lithospermic acid are the six major active constituents in Danshen injection. In this study, a rapid, sensitive and specific liquid chromatographic-electrospray ionization-mass spectrometry method for the simultaneous quantitative determination of these compounds in rat plasma was developed. After a single step of liquid-liquid extraction with ethyl acetate, they were eluted by a Hypersil C18 column (5 µm, i.d. 4.6 × 200 mm) within 4 min with a mobile phase consisting of acetonitrile and 0.1% formic acid water solution (35:65, v/v). The assay was linear in the concentration range of 0.05-10 µg mL(-1). Absolute recoveries were above 60%. The precisions and accuracies determined within three consecutive days were within acceptable limits. The method was successfully applied to a pharmacokinetic study in rats after an intravenous administration of Danshen injection.

Topics: Animals; Benzaldehydes; Benzofurans; Caffeic Acids; Catechols; Chromatography, High Pressure Liquid; Chromatography, Liquid; Cinnamates; Depsides; Drug Stability; Drugs, Chinese Herbal; Injections, Intravenous; Lactates; Liquid-Liquid Extraction; Male; Mass Spectrometry; Plant Preparations; Rats; Reference Standards; Rosmarinic Acid; Salvia miltiorrhiza; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization

To investigate the pharmacokinetic interactions induced by content variation of the main water-soluble components of Danshen injection in rats.. Intravenous Danshen injection (control) or Danshen injection with danshensu (DSS), protocatechuic aldehyde (PAL), salvianolic acid A (Sal A) or salvianolic acid B (Sal B) were administered to female Sprague Dawley rats. Plasma concentrations of DSS, Sal A, PAL and its oxidative metabolite protocatechuic acid (PA) were analyzed simultaneously with LC-MS/MS; concentrations of Sal B were determined by the LC-MS method. Non-compartmental pharmacokinetic parameters were calculated and compared for identifying the pharmacokinetic interactions among these components.. Compared with the control group, the DSS, Sal A, and Sal B groups had significant increases in AUC(0-infinity) in response to elevated concentrations of PAL (by 78.1%, 51.0%, and 82.9%, respectively), while the clearances (CL) were markedly reduced (by 42.5%, 32.9%, and 46.8%, respectively). Similarly, Sal A increased the AUC(0-infinity) of DSS and Sal B (26.7% and 82.4%, respectively) and substantially decreased their clearances (21.4% and 45.6%, respectively). In addition, the pharmacokinetics of DSS and Sal B were significantly affected by the content variation of the other major components; the AUC(0-infinity) increased by 45.1% and 52.1%, respectively, the CL dropped by 29.6% and 27.1%, respectively, and the T(1/2) was decreased by 22.0% and 19.6%, respectively.. Complex, extensive pharmacokinetic interactions were observed among the major water-soluble constituents in the Danshen injection. The content variation of PAL had the most significant effect on the pharmacokinetic behaviors of other major constituents. Furthermore, the pharmacokinetics of DSS and Sal B were the most susceptible to the content change of other components.

Topics: Animals; Benzaldehydes; Benzofurans; Caffeic Acids; Catechols; Drugs, Chinese Herbal; Female; Lactates; Phenanthrolines; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza

To study the chemical constituents in the root of Salvia yunnansis.. Compounds were isolated and purified by Diaion HP20, Sephadex LH - 20, ODS chromatography. Their structures were determined by spectral analysis and chemical evidence.. Twelve compounds were isolated and identified from the root of S. yunnansis protocatechaldehyde (1), caffeic acid (2), ferulic acid (3), rosmarinic acid (4), salvianolic acid A (5), salvianolic acid C (6), lithospermicacid (7), lithospermicacid B (8), 9'-methyl lithospermate B (9), 9"'-methyl lithospermate B (10), 9',9'''-dimethyl lithospermate B (11), 9'-ethyl lithospermate B (12).. The compounds 1, 2, 3, 5, 6, 9, 10, 11 and 12 were first isolated from S. yunnanensis.

Topics: Benzaldehydes; Caffeic Acids; Catechols; Chromatography; Drugs, Chinese Herbal; Flavonoids; Lactates; Phenols; Plant Roots; Plants, Medicinal; Polyphenols; Resins, Synthetic; Salvia

Salvianolic acid A (1) is one of the active components from Salvia miltiorrhiza, which was found to suppress the growth of mouse tumors. S-3-1 (a 2-allyl-3,4-dihydroxybenzaldehyde, 2) is a synthetic intermediate of a salvianolic acid A derivative with strong inhibitory effects on the growth of cancer cells in vitro. The inhibitory effects of 2 on tumor growth and its molecular targets were studied. 2 significantly suppressed the growth of mouse Lewis lung carcinoma, S180 sarcoma and H22 hepatic carcinoma in a dose-dependent manner. With a simple scrape-loading dye transfer method, 20 microg/ml of 2 was found to significantly enhance gap junction intercellular communication (GJIC) in human pancreatic adenocarcinoma PaCa Cells, human lung epithelial carcinoma W1-38 cells and human lung adenocarcinoma A549 cells, but 2 had no marked effect on GJIC in human colon cancer CACO2 cells. With Northern blot analysis, 2 was found to inhibit the expression of c-myc gene in A549 cells and have no marked effect on H-ras oncogene expression, and increase the cellular P53 mRNA contents, though it did not affect the expression of RB tumor suppressor gene. 2 also suppressed the P46 (JNK/SAPK) expression in A549 cells. Western blot analysis was applied to visualize the P21ras protein. Results shows that 2 at concentrations ranging from 10 to 20 microg/ml decreases the contents of the membranous P21ras and total P21ras and increases the contents of cytosolic P21ras protein in a time-dependent manner. However, 2 had no significant effects on farnesyl protein transferase activities at the concentrations that could efficiently decrease the membranous P21ras content. This suggested that 2 might suppress tumor growth partly through enhancement of GJIC and reversion of the transformed phenotypes. The other mechanisms may be that 2 can suppress the overexpression of c-myc oncogene, inhibit the function of Ras oncoprotein, increase the expression of P53 tumor suppressor gene and interrupt P46-associated mitogen-activated pathway other than farnesylation of Ras protein.

Topics: Allyl Compounds; Animals; Benzaldehydes; Blotting, Western; Caco-2 Cells; Caffeic Acids; Catechols; Cell Division; Dose-Response Relationship, Drug; Gene Expression Regulation, Neoplastic; Genes, myb; Genes, p53; Humans; Lactates; Liver Neoplasms; Lung Neoplasms; Mice; Mitogens; Phenotype; Sarcoma; Tumor Cells, Cultured

A new analytical method for the separation and determination of seven aqueous depsides in salvia miltiorrhiza by HPTLC has been developed. The seven depsides are protocatechualdehyde, caffeic acid, methyl rosmarinate, rosmarinic acid, salvianolic acid A, B and C. Using chloroform-ethyl acetate-benzene-formic acid(2.4:2:1:0.6) as developing solvent A, protocatechualdehyde was separated; using chloroform-ethyl acetate-benzene-formic acid-methanol (1.5:2:1:1:0.1) as developing solvent B, the other six constituents were well separated. The aqueous depsides were detected at the wave lengths of lambda S = 300 nm and lambda R = 240. This method is simple, rapid, sensitive and accurate. The contents of seven depsides in several Salvia species were determined.

Topics: Benzaldehydes; Caffeic Acids; Catechols; Chromatography, Thin Layer; Cinnamates; Densitometry; Depsides; Drugs, Chinese Herbal; Lactates; Plant Extracts; Rosmarinic Acid; Salvia miltiorrhiza

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