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In-vitro inhibition of human erythrocyte acetylcholinesterase by salvia lavandulaefolia essential oil and constituent terpenes.

Sage (Salvia spp) is reputed in European herbal encyclopaedias to enhance memory, and current memory-enhancing/anti-dementia drugs are based on enhancing cholinergic activity by inhibiting cholinesterase. In this study the effects of Salvia lavandulaefolia Vahl. (Spanish sage) essential oil and some of its constituent terpenes on human erythrocyte acetylcholinesterase were examined in-vitro. The main constituents in the essential oil batch used for analysis of cholinesterase inhibition were camphor (27%), 1,8-cineole (13%), alpha- and beta-pinene (10-15%) and bornyl acetate (10%) with other minor constituents (1% or less) including geraniol, limonene, linalool, terpineol and gamma-terpinene. Using the Ellman spectrophotometric method, kinetic analysis was conducted on the interaction of the essential oil and the main monoterpenoids, camphor, 1,8-cineole and alpha-pinene. IC50 values were obtained for the essential oil, 1,8-cineole and alpha-pinene and were 0.03 microL [corrected] mL(-1), 0.67 mM and 0.63 mM, respectively. Camphor and other compounds tested (geraniol, linalool and gamma-terpinene) were less potent (camphor IC50: >10mM). The essential oil, alpha-pinene, 1,8-cineole and camphor were found to be uncompetitive reversible inhibitors. These findings suggest that if the inhibitory activity of the essential oil is primarily due to the main inhibitory terpenoid constituents identified, there is a major synergistic effect among the constituents. Since no single constituent tested was particularly potent, it remains to be determined whether these in-vitro cholinesterase inhibitory activities are relevant to in-vivo effects of the ingestion of S. lavandulaefolia essential oil on brain acetylcholinesterase activity.

Acetylcholinesterase↗

Comparative study of seventeen Salvia plants: aldose reductase inhibitory activity of water and MeOH extracts and liquid chromatography-mass spectrometry (LC-MS) analysis of water extracts.

The dry root and rhizome of Salvia miltiorhiza (Lamiaceae) are used as a crude drug Danshen, while those of S. deserta (Xinjiang-Danshen) are mixed in Danshen at Xinjiang province when the former is in short supply. The water and MeOH extracts of S. deserta showed strong aldose reductase (AR) inhibitory activity, and their active constituents were determined to be polar compounds different from "tanshinones" of S. miltiorhiza, i.e., lithospermic acid B (1), salvianolic acid K (2), salviaflaside (3), and rosmarinic acid (4) (IC50, 2.63-3.91 microM). We also examined the AR inhibitory activity of water and MeOH extracts of seventeen Salvia plants, including ten species of Danshen resources (S. bowleyana, S. deserta, S. miltiorhiza, S. miltiorhiza var. miltiorhiza f. alba, S. paramiltiorhiza, S. paramiltiorhiza f. purpureo-rubra, S. przewalskii, S. przewalskii var. mandarinorum, S. sinica f. purpurea, S. trijuga), and their water extracts were also analyzed by liquid chromatography-mass spectrometry (LC-MS). The results indicated that there were four types with regard to the AR inhibitory activity and three types with regard to the amount of 1. Ten species used as Danshen resources showed good correlation between the AR inhibitory activity and the morphological classification. However, the intensities of their AR inhibitory activity varied, and they contained 1 in varying amounts. These facts suggested that the ten species were not the same, and thus their use as a Danshen resource should be based on their activity and/or active constituents.

Aldehyde Reductase↗

Cardiac effects of salvia miltiorrhiza/dalbergia odorifera mixture, an intravenously applicable Chinese medicine widely used for patients with ischemic heart disease in China.

Chronotropic, inotropic and coronary vasodilator actions of the clinically available ampoule preparation of Salvia miltiorrhiza/Dalbergia odorifera mixture were examined using canine isolated, blood-perfused heart preparations. The mixture slightly decreased the sinoatrial rate and significantly increased coronary blood flow, but hardly affected the developed tension of the papillary muscle. The effect on coronary blood flow was induced by at least a 10-fold smaller dose than that which induced the chronotropic effect. These results were quite similar to those of a typical calcium channel blocker, verapamil, used in a previous study, suggesting that the Salvia miltiorrhiza/Dalbergia odorifera mixture may have potential as an anti-anginal drug.

Animals↗

Screening for antioxidant properties of Salvia reflexa hornem.

The antioxidant properties of the wild growing sage species, Salvia reflexa Hornem., were investigated. The presence of superoxide (O(2)(.-)) and hydroxyl ((.)OH) radicals, malonyldialdehyde (MDA), reduced glutathione (GSH) and total flavonoids were observed in the above-ground parts of plant, as well as activities of the antioxidant enzymes superoxide-dismutase (SOD) and peroxidase (P-ase). The potential antioxidant activity of the methanol:water extract has been assessed based on scavenging activity of stable DPPH free radicals. By the means of TLC and LC/MS a screening for secondary plant products was performed. Significant quantities of O(2)(.-), (.)OH and MDA were observed. Thus, this species exhibited high SOD and P-ase activities as well as a content of total flavonoids. The dominant naturally occurring compound was rosmarinic acid. The results obtained suggested a high antioxidant activity of Salvia reflexa Hornem.

Antioxidants↗

Triterpenoids and other compounds from Salvia roborowskii Maxim.

Two new triterpenoids, 2 alpha-hydroxy-3 beta-methoxyurs-12-en-28-oic acid and 3 alpha-hydroxy-2 alpha-methoxyurs-12-en-28-oic acid as well as 25 known compounds were isolated from the whole plant of Salvia roborowskii Maxim. Their structures were elucidated by means of spectral data and chemical transformation. This is the first report on the chemical constituents of this plant. The presence of eugenyl beta-D-glucopyranoside 22 in a plant of the genus Salvia is also reported for the first time.

Acetylation↗

[Thin layer chromatographic studies on radix Ginseng, fructus Schisandrae and radix Salviae miltiorrhizae in yixinfumai granules].

The main constituents of Radix Ginseng, Fructus Schisandrae and Radix Salviae Miltiorrhizae in Yixinfumai Granules were studied by TLC. It was found that ginsenoside got hydrolyzed while the drugs were being boiled together. The main constituents of Fructus Schisandrae were schizandrol A and B; and those of Radix Salviae Miltiorrhizae were tanshinone and protocatechuic aldehyde.

Chromatography, Thin Layer↗

The water-soluble component of Salvia miltiorrhiza Bge can promote macrophage synthesis of C4 in vivo and in vitro.

A water-soluble component extracted from Salvia miltiorrhiza Bge has been used for many years in traditional Chinese medicine ('huo xie hua yu') for the treatment of the hyperviscosity syndrome. We demonstrate here that this extract promotes macrophage proliferation in vitro and enhances macrophage synthesis of the C4 component of complement. No increase in synthesis of other complement components by macrophages was observed. In a group of 20 patients with the hyperviscosity syndrome treated with an aqueous extract of Salvia, serum levels of C4 rose significantly over the course of treatment (mean pretreatment level 37.4 mg%, mean posttreatment level, 43.5 mg%; p < 0.01). The rise in C4 levels closely correlated with the disappearance of the patients' clinical symptoms.

Blood Viscosity↗

[Royleanones in the roots of Salvia officinalis L. of domestic provenance and their antimicrobial activity].

The reported investigation of the constituents of the petroleum ether extract of the root of Salvia officinalis L. confirmed the presence of the following diterpene quinones: 12.hydroxy-8, 12-abietadiene-11, 14-dione (royleanone), 7 alpha, 12-dihydroxy-8.12-abistadiene-11-14-dione (horminone) and 7 alpha-acetoxy-12-hydroxy-8, 12-abietadiene-11,14-dione (7-O-acetylhorminone). The compounds were identified on the basic of the interpretation of results of spectral analysis. Isolated royleanones show antimicrobial activity against gram-positive bacteria (Staphylococcus aureus). A study of abietate diterpenoids in the root of Salvia officinalis L. of Slovak provenance has been performed for the first time.

Abietanes↗

Clinical non-inferiority trial on treatment of coronary heart disease angina pectoris of Xin-blood stasis syndrome type with lyophilized Salvia salt of lithospermic acid powder for injection.

OBJECTIVE: To evaluate the effectiveness and safety of lyophilized Salvia salt of lithospermic acid powder for injection (SSLA) in treating coronary heart diseases angina pectoris (CHD-AP) of Xin-blood stasis syndrome type, and to conduct the non-inferiority trial with Danshen injection (DSI) as positive control. METHODS: An non-inferiority clinical layered, segmented, randomized, and blinded trial on three parallel and multiple centered groups was conducted in 480 patients with stable effort angina grade I, II and III, who had two or more times of attack every week. The 240 patients in test group A were treated with SSLA 200 mg added in 250 ml of 5% glucose solution for intravenous dripping every day; the 120 patients in test group B were treated with SSLA but the dosage doubled; and the 120 patients in the control group were treated with DSI 20 ml daily in the same method as SSLA was given. The clinical effectiveness and safety were evaluated after the patients were treated for 14 days. RESULTS: The results showed that the markedly effective rate in test groups A, B and control group was 37.45%, 36.75% and 30.09% respectively, while the total effective rate in them was 88.09%, 89.74% and 67.26% respectively. Statistical significance was shown in comparisons of the therapeutic effect between control group with test group A and test group B, with that in the two test groups superior to that in the control group, and non-inferiority trial showed eligibility (P < 0.01). Adverse reaction appeared in 8 patients in the test groups and 2 in the control group. CONCLUSION: SSLA has definite therapeutic effect in treating patients with CHD-AP, with its effect not inferior to that of DSI, and no evident toxic-adverse reaction.

Adolescent↗

[Review of germplasm resources studies on Salvia miltiorrhiza].

Research development of germplasm resources of Salvia miltiorrhiza was reviewed in the aspects of species resource, geographic distribution and varieties, biological characters and culture varieties, and cell and tissue culture. It was suggested that more attention be paid to waters-soluble constituents when evaluating the quality of the medicine, and much more basic studies be done in the course of establishing standard plant base for high quality herb and in the aspect of new variety selection.

Abietanes↗

Rapid screening of antioxidant activity of sage (Salvia officinalis L.) extracts obtained by supercritical carbon dioxide at different extraction conditions.

Sage herb (Salvia officinalis L.) was extracted at supercritical fluid extraction (SFE) conditions with carbon dioxide at different parameters and the extracts tested on their antioxidant activity (AA). SFE of sage herb at 35 MPa pressure was found to be an effective method to obtain pure extracts. The yields of the extracts were substantially increased by using 1% of entrainer solvent ethanol. The fractionation of sage extract was a complex procedure in terms of extract distribution between separators operating at various pressure and temperature conditions. It was also proved by testing the AA of the extracts in rapeseed oil. The effect of the extracts on the rapeseed oil weight gain varied in a wide range (from 'very low' to 'high') depending on the fractionation conditions. Preliminary results showed that to obtain more effective antioxidant fractions separation steps should be started at 10 MPa lower pressure than that used for the extraction.

Antioxidants↗

Determination of Danshensu, a major active compound of Radix Salvia miltiorrhiza in dog plasma by HPLC with fluorescence detection.

A simple and sensitive high performance liquid chromatography method has been developed for the determination of Danshensu (3, 4-dihydroxyphenyllactic acid) in dog plasma. Plasma samples were extracted with ethyl acetate. Analysis of the extracts was performed on a reversed-phase column with an aqueous phosphate buffer-acetonitrile (93:7, v/v) mobile phase, and 4-hydroxybenzoic acid was used as the internal standard. Fluorescence detection at 285 nm (excitation) and 320 nm (emission) was employed. Standard curves were linear in the range from 0.125 to 11.3 microg/mL (regression coefficient r > 0.993) on three different days. Mean recovery was determined as 96.4% by analysis of plasma standard containing 0.63, 5.65 and 11.3 microg/mL of Danshensu. The inter-day precision (RSD) ranged from 3.4 to 8.6% at concentrations of 0.125, 1.88, 6.28 and 11.3 microg/mL, and the intra-day precision was better than 7.2%. The detection and quantitation limits were 0.063 and 0.125 microg/mL, respectively. This validated assay was applied to the determination of Danshensu concentration in dog plasma after oral administration of Radix Salvia miltiorrhiza extracts.

Administration, Oral↗

HPLC determination and pharmacokinetic studies of salvianolic acid B in rat plasma after oral administration of Radix Salviae Miltiorrhizae extract.

A precise and reproducible HPLC method has been established and validated for determination of salvianolic acid B (SalB) in rat plasma after oral administration of Radix Salviae Miltiorrhizae extract. Liquid-liquid extraction was adopted for the sample preparation. Separation was accomplished on a C(18) column with a linear gradient elution consisting of acetonitrile and aqueous phosphoric acid. Ultraviolet detection was at 280 nm. The method was validated over the concentration range 10.8-259.4 microg/mL using 1 mL of plasma. The assay was linear over this concentration range with a coefficient of variation less than 7%. The extraction recovery of SalB was within the range 71-83% with RSD 11%. The mean recovery of the internal standard was 84% (n = 6) with RSD of 5.6%. This method is suitable to determine SalB in plasma and to investigate the pharmacokinetics of SalB.

Acetonitriles↗

Characterization and bioscreening of a new triterpenoid and a flavanone isolated from Salvia nubicola.

A new epimeric lupane triterpenoid, 3alpha-hydroxy-20-oxo-30-norlupane (1), and a new flavanone (nubatin; 3) have been isolated from Salvia nubicola collected from Quetta, Pakistan. The structures of these constituents were elucidated with the aid of spectroscopic techniques, including two-dimensional NMR and NOE experiments. Both the metabolites were found to be moderately bio-active.

Anti-Bacterial Agents↗

Novel rearranged abietane diterpenoids from the roots of Salvia sahendica.

Two naphthoquinone diterpenoids, 1 and 2, one tricyclic, and one tetracyclic rearranged abietane ('4,5-seco-10,5-friedo-abietane') diterpenoids, 3 and 4, respectively, together with horminone (5) have been isolated from the roots of Salvia sahendica. Compounds 2 and 3 are new, and the 13C-NMR assignment for compound 4 was modified using ' Heteronuclear Multiple-Bond Correlation' (HMBC) spectroscopic data. The structures of the compounds have been established by using different spectral data including 1D- and 2D-NMR, IR, UV, and MS. The elemental composition for the major peaks of 3 and 4 were determined by ' High-Resolution Electron Impact Mass Spectrometry' (HR-EI-MS). The relative configurations of the new compounds were determined by 1H-NMR and 'Rotating-Frame NOES' (ROESY) spectroscopy. Compounds 1, 2, and 5 showed antifungal activities when tested on Blakeslea trispora. Lapachol, a prelynated naphthoquinone, was used as a positive control. The biological activities of the related naphthoquinones and abietane diterpenoids were discussed.

Abietanes↗

Triterpenoids from Salvia wagneriana.

From the exudate of Salvia wagneriana Polak, beside the known ursolic acid, two pentaoxygenated triterpenoids were isolated. Their structures were determined as 3-oxo-11alpha,19beta,20,22beta-tetrahydroxy-lupane (1) and 3beta,11alpha,19beta,20,22beta-pentahydroxy-lupane (2) using a combination of one- and two-dimensional NMR techniques.

Chromatography, Thin Layer↗

Salvia miltiorrhiza inhibits intimal hyperplasia and monocyte chemotactic protein-1 expression after balloon injury in cholesterol-fed rabbits.

Antioxidants that prevent low density lipoproteins (LDL) from oxidation may inhibit atherosclerosis and post-angioplasty restenosis. Salvia miltiorrhiza (SM) has been shown to inhibit LDL oxidation and reduce atherosclerosis in cholesterol-fed rabbits. The effects of SM on neointimal hyperplasia and monocyte chemotactic protein-1 (MCP-1) expression after balloon injury were studied. Male New Zealand white rabbits were fed a 2% cholesterol diet together with daily SM (4.8 gm/kg body wt.) treatment (SM; n=10) or without SM as a control (C; n=9) for 6 weeks. Probucol-treated (0.6 gm/kg body wt.) rabbits (P; n=9) were used as a positive control group. A balloon injury of the abdominal aorta was performed at the end of the third week. Aortas were harvested at the end of 6 weeks. The plasma cholesterol levels were lowered in SM group. The neointimal hyperplasia in abdominal aortas was significantly inhibited in SM group [neointima/media area ratio: 0.63+/-0.05 (SM) versus 0.78+/-0.05 (C); P < 0.05] and in P group [0.45+/-0.02 (P) versus 0.78+/-0.05 (C); P < 0.05] when compared with C group. SM treatment significantly reduced MCP-1 mRNA and protein expression in balloon-injured abdominal aorta. These inhibitory effects on intimal response after balloon injury might be attributed to antioxidant capacity and cholesterol lowering effect of SM. SM treatment may offer some protection against post-angioplasty restenosis.

Angioplasty, Balloon↗