A combination extract of kaffir lime, galangal, and lemongrass maintains blood lipid profiles, hepatocytes, and liver mitochondria in rats with nonalcoholic steatohepatitis.

Kaffir lime leaves and the rhizomes of galangal and lemongrass are the main ingredients in several Thai foods with desirable medicinal effects. Based on their beneficial activities, this study aimed to indicate the chemical properties and in vivo efficacy of a combination of the herbs at a 1:2:1 ratio in a water extract form. Its volatile constituents were analyzed by gas chromatography coupled with mass spectrometer, which mainly consists of eucalyptol, citronellal, and citral. Clinicohistopathological and electron microscopic studies demonstrated that the extract corrected blood cholesterol, LDL, HDL, and triglyceride levels similarly as simvastatin treatment in association with its antioxidative properties, as indicated by the levels of superoxide dismutase and malondialdehyde in serum and the increment of nuclear factor erythroid 2-related factor 2 levels in hepatocytes. Hepatitis was significantly less severe in rats fed the extract for 14 days than in simvastatin-treated rats. Regarding its immunomodulatory properties, the extract also accelerated hepatic resolution from steatohepatitis during hypercholesterolemia as indicated by the upregulation of vimentin, cytokeratin, and CD206+. Interestingly, liver mitochondria were also preserved in hypercholesterolemic rats treated with the extract in relation to their architecture and the expression of haloacid dehalogenase-like hydrolase domain-containing protein 3 as well as metabolic and energy regulators. Therefore, the study concluded that the water extract of kaffir lime leaves and the rhizomes of galangal and lemongrass has beneficial effects on blood cholesterol, the severity of steatohepatitis, and the maintenance of mitochondrial architecture via its antioxidative and immunomodulatory activities.

A new 3,4-seco-cycloartane from the leaves of Hopea odorata Roxb.

A new 3,4-seco-cycloartane, identified as (24R,25S)-dihydroxy-26-O-nonadecylcarbonyloxy-3,4-secocycloarta-4(28)-en-3-oic acid (1), has been isolated from the leaves of Hopea odorata Roxb. (Dipterocarpaceae), together with the rare 3,4-seco-cycloart-4(28),24-diene-3,26-dioic acid (2 or abiesatrine J) and six other known compounds (3-8). Their structures were elucidated on the basis of both chemical and spectroscopic methods.

Effect of Short-Term Oral Administration of Phikud Navakot in Rats.

BACKGROUND: Phikud Navakot (PN), composed of nine herbs and used as a main component of Yahom Navakot, is used in traditional Thai medicine against dizziness and fainting. OBJECTIVE: To investigate the effects of PN on blood pressure, heart rate (HR), and antioxidant properties on male Sprague Dawley rats. MATERIAL AND METHOD: All rats were weighted everyday in the morning, after that, PN (10, 50, 100, 200 and 400 mg/kg BW) were given oroesophageal feeding for seven days. Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and HR were measured once per two days. At the end of the experiment, the blood was taken for determination of biochemical and hematological parameters, and lipid peroxidation in serum. The heart was immediately removed for Western blot analysis. RESULTS: SBP DBP and MAP of rats were transiently increased after 1 day of PN (100 mg/kg BW) treatment. Meanwhile, HR did not change throughout the experiment. PN (400 mg/kg BW) significantly increased (p< 0.05) the percentage of neutrophils in blood after 7 days of administration. PN treatment has no effect on biochemical parameters and peroxidation of lipid. In addition, ingestion of PN (100 mg/kg BW) significant increased (p < 0.05) HO-1 expression, but did not change ERK1/2 and Bax/Bcl-2 ratio when compared with the control group. CONCLUSION: The results may possibly support the use of PN for prevention and/or alleviation of cardiovascular disorders, caused by reactive oxygen species. However, long-term treatment of PN has to be further studies.

Effects of Phikud Navakot Extract on Myocardial Ischemia/Reperfusion Injury in Rats.

BACKGROUND: Phikud Navakot (PN) is a set of nine medicinal plants and the main ingredient of "Yahom Navakot", a traditional Thai herbal formula for treatment of cardiovascular symptoms. OBJECTIVE: To investigate the cardioprotective effects of PN on myocardial ischemia/reperfusion (IR) in male Sprague Dawley rats. MATERIAL AND METHOD: Rats were randomly divided into 7 groups: sham, IR, and IR orally pretreated with PN (10, 50, 100, 200, and 400 mg/kg B W)for 7 days. After treatment, IR induction was performed by left coronary artery (LCA) ligation for 30 min, followed by reperfusion for 24 h. At the end of the experiment, blood was collected for hematological and biochemical parameters, and hearts were immediately removed for histopathological examination and Western blot analysis. RESULTS: IR induction caused ST elevation in the electrocardiogram and an increase in serum troponin I (TnI), confirming myocardial damage. In addition, histopathological changes of ischemic myocardium showed inflammation, infiltration, and edema. Oral administration of PN (10, 50, 100, 200, and 400 mg/kg BW) for 7 days prior to IR simulation showed no change on serum TnI and histopathology ofcardiac tissues, when compared to IR group. However Western blot analysis showed that IR rats pretreated with PN (10 mg/kg BW) significantly increased (p < 0.05) pERK/ERK ratio, meanwhile pretreated with PN (50-200 mg/kg BW) up-regulated (p < 0.05) the protein expression of HO-1, when compared with IR group. CONCLUSION: The present study implied that 7-day pretreatment of PN failed to protect cardiac tissues against IR injury induced by LCA ligation. Investigation at molecular level found however that PN up-regulated the expression of protective proteins pERK/ERK ratio and HO-1 in cardiac tissues, suggesting molecular mechanism of PN in cardioprotection against IR injury.

Synergistic antioxidant action of Phikud Navakot ameliorates hydrogen peroxide-induced stress in human endothelial cells.

BACKGROUND: Phikud Navakot (PN), a combination of nine herbs, has been used traditionally in Thai medicinal formulas to relieve circulatory disorder. The present study aimed to compare the synergistic antioxidant efficacy and toxicity of the hydroethanolic and water extracts of PN at cellular level. METHODS: PN and its nine herbs were extracted with either 50% ethanol or water. All extracts were tested for in vitro antioxidant potential using standard antioxidant assays. Evaluation of cytotoxicity, genotoxicity, and intracellular reactive oxygen species were performed using human endothelial ECV304 cells. RESULTS: Antioxidant assays in cell-free systems showed that the hydroethanolic extract of PN scavenged superoxide, hydroxyl, nitric oxide radicals, and hydrogen peroxide more effectively than its water extract. Combination indices were calculated to show that the ingredients of the hydroethanolic extract acted synergistically to exhibit antioxidant activities against all tested radicals, whereas, in the case of water extract, this effect was observed only against 2,2-diphenyl-1-picrylhydrazyl, superoxide, and hydroxyl radicals. A cell-based assay also revealed that the hydroethanolic extract concentration-dependently attenuated hydrogen peroxide-induced stress more effectively than the water extract. At the antioxidant and cytotoxic concentrations of both extracts, no genotoxicity was found. CONCLUSION: Our findings demonstrate that the synergistic antioxidant action of PN ameliorates endothelial stress, which may provide some clues for understanding the traditional use of PN for the treatment of circulatory disorder. Additionally, the selection of a suitable solvent for the extraction of PN herbal combination is essential for maximal efficacy and safety.

Quantitative determination of sibutramine in adulterated herbal slimming formulations by TLC-image analysis method.

A simple thin layer chromatographic (TLC)-image analysis method was developed for rapid determination and quantitation of sibutramine hydrochloride (SH) adulterated in herbal slimming products. Chromatographic separation of SH was achieved on a silica gel 60 F(254) TLC plate, using toluene-n-hexane-diethylamine (9:1:0.3, v/v/v) as the mobile phase and Dragendorff reagent as spot detection. Image analysis of the scanned TLC plate was performed to quantify the amount of SH. The polynomial regression data for the calibration plots showed good linear relationship in the concentration range of 1-6 µg/spot. The limits of detection and quantitation were 190 and 634 ng/spot, respectively. The method gave satisfactory specificity, precision, accuracy, robustness and was applied for determination of SH in herbal formulations. The contents of SH in adulterated samples determined by the TLC-image analysis and TLC-densitometry were also compared.

Effects of Phikud Navakot extract on vascular reactivity in the isolated rat aorta.

The aim of the present study is to investigate the effect of standardized Phikud Navakot extract (NVKE) on aortic rings from male Sprague Dawley rats. Changes in tension were measured using an isometric force transducer and recorded on the PowerLab recording system. Vasorelaxant effect of NVKE was examined in the presence of indomethacin (10 microM), N(G)-nitro L-arginine methyl ester (L-NAME, 300 microM), methoxamine (0.1-300 microM), carbachol (1 nanoM-30 microM), or sodium nitroprusside (0.1 nanoM-10 microM). The results showed that NVKE (1-300 microg/mL) caused vasorelaxation in a concentration-dependent manner with a pEC50 value of 4.27 +/- 0.24 and R(max) of 67.7 +/- 13.9%. Pretreatment with indomethacin or L-NAME did not affect NVKE-induced vasorelaxation. However, co-incubation of indomethacin and L-NAME significantly reduced (p < 0.05) vasorelaxation to NVKE (100 microg/mL). Pre-treatment with NVKE significantly decreased (p < 0.05) endothelium-dependent relaxations to carbachol (R(max) = 52.90 +/- 14.3%), but not to sodium nitroprusside. Moreover contractions to methoxamine were unaffected after pretreatment with NVKE. The present study suggested that NVKE decreased vasorelaxation to carbachol in the rat aorta, which may exert at least against muscarinic receptors. These findings support the use of Phikud Navakot, a major ingredient of Yahom Navakot, against dizziness and fainting.

Thai plants from Doi Tung: brine shrimp lethality, antioxidative activity and combination effect with L-ascorbic acid.

Methanolic extracts of 18 Thai plants from Doi Tung, the north of Thailand, were examined for brine shrimp lethality and antioxidative activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. In brine shrimp lethality, the highly toxic plant extracts were Mitrephora wangii (LC(50) 14.8 µg mL(-1)) and Hydrocotyle javanica (LC(50) 13.3 µg mL(-1)). The extracts of Sageretia cordifolia, Ulmus lancaefolia and Acer chiangdaoense exhibited highly antioxidative activities with IC(50) values of 6.7, 8.1 and 9.8 µg mL(-1), respectively. Moreover, the extracts of S. cordifolia, U. lancaefolia and A. chiangdaoense have shown slightly synergistic effects (combination index; CI < 1) with L-ascorbic acid on DPPH radicals. A number of plants demonstrated either brine shrimp lethality or free radical scavenging activity.

Geraniinic acid derivative from the leaves of Phyllanthus reticulatus.

Chemical constituents as well as cytotoxic and insecticidal activity of the crude methanol extract from the leaves of Phyllanthus reticulatus Poir. (Euphorbiaceae) were investigated. (5R*,6R*)-4,6-Dimethoxycarbonyl-5-[2',3',4'-trihydroxy-6'-(methoxycarbonyl) phenyl]-5,6-dihydro-2H-pyran-2-one (1) along with 3,4,3'-tri-O-methylellagic acid, and methyl gallate were isolated from the dichloromethane extract. Determination of their structures was based on spectroscopic analysis. Compound 1 possessed a very weak insecticidal activity against Spodoptera frugiperda (Sf9) with an IC(50) value of 27.27 microg/mL.

Effects of Curcuma spp. on P-glycoprotein function.

The effects of Curcuma longa (khamin chan) and Curcuma sp. "khamin-oi" (khamin-oi), as well as isolated major curcuminoids on intestinal P-gp functions were evaluated in vitro. The accumulation of R123 in Caco-2 cells was increased and the R123 efflux ratios were significantly decreased by both Curcuma longa and Curcuma sp. "khamin-oi" extracts, indicating their roles on efflux transporters. The a-b transport of daunorubicin was increased by curcumin, demethoxycurcumin and bisdemethoxycurcumin while the b-a transport was significantly decreased by curcumin and demethoxycurcumin. However, calcein-AM uptake into the human P-gp overexpression cell line, LLC-GA5-COL300, was increased by curcumin and demethoxycurcumin in a concentration-dependent manner but not affected by bisdemethoxycurcumin. These results show that curcumin and demethoxycurcumin could inhibit P-gp but bisdemethoxycurcumin may modulate the function of other efflux transporters such as MRP. Taken together, the information may indicate the impact of Curcuma longa and Curcuma sp. "khamin-oi" on pharmacokinetics of orally administered drugs that are P-gp substrates.

Stability of barakol under hydrolytic stress conditions and its major degradation product.

The aim of the present study was to investigate the stability of barakol, an anxiolytic constituent extracted from leaves of Senna siamea (Lam.) Irwin & Barneby (syn. Cassia siamea Lam.), under the International Conference on Harmonisation suggested conditions using HPLC with photodiode array detection. Extensive degradation of barakol was found to occur under alkaline conditions through base-catalyzed hydrolysis. Mild degradation of barakol was observed under thermal and oxidative stress while it was stable under acidic conditions. The reaction rate constants (Kobs) of barakol degradation under alkaline conditions at pHs 12 and 13 were 3.0x10(-5) and 9.6x10(-3) min(-1), respectively. The activation energy according to the Arrhenius plot was calculated to be 26.9+/-3.3 kcal/mol at pH 13 and temperatures between 12 and 51 degrees C. The major degradation product of barakol under both alkaline and thermal stress conditions was characterized by LC-MS and NMR as cassiachromone.

Application of Scion image software to the simultaneous determination of curcuminoids in turmeric (Curcuma longa).

INTRODUCTION: Curcumin, desmethoxycurcumin and bisdesmethoxycurcumin are bioactive constituents of turmeric (Curcuma longa). Owing to their different potency, quality control of turmeric based on the content of each curcuminoid is more reliable than that based on total curcuminoids. However, to perform such an assay, high-cost instrument is needed. OBJECTIVE: To develop a simple and low-cost method for the simultaneous quantification of three curcuminoids in turmeric using TLC and the public-domain software Scion Image. METHODOLOGY: The image of a TLC chromatogram of turmeric extract was recorded using a digital scanner. The density of the TLC spot of each curcuminoid was analysed by the Scion Image software. The density value was transformed to concentration by comparison with the calibration curve of standard curcuminoids developed on the same TLC plate. RESULTS: The polynomial regression data for all curcuminoids showed good linear relationship with R(2) > 0.99 in the concentration range of 0.375-6 microg/spot. The limits of detection and quantitation were 43-73 and 143-242 ng/spot, respectively. The method gave adequate precision, accuracy and recovery. The contents of each curcuminoid determined using this method were not significantly different from those determined using the TLC densitometric method. CONCLUSION: TLC image analysis using Scion Image is shown to be a reliable method for the simultaneous analysis of the content of each curcuminoid in turmeric.

Spectrophotometric method for quantitative determination of total anthocyanins and quality characteristics of roselle (Hibiscus sabdariffa).

A simple, rapid and cost-saving method for the determination of total anthocyanins in roselle has been developed. The method was based on pH-differential spectrophotometry. The calibration curve of the major anthocyanin in roselle, delphinidin 3-sambubioside (Dp-3-sam), was constructed by using methyl orange and their correlation factor. The reliability of this developed method was comparable to the direct method using standard Dp-3-sam and the HPLC method. Quality characteristics of roselle produced in Thailand were also reported. Its physical quality met the required specifications. The overall chemical quality was herein surveyed for the first time and it was found to be the important parameter corresponded to the commercial grading of roselle. Total contents of anthocyanins and phenolics were proportional to the antiradical capacity.

Antioxidative and neuroprotective activities of extracts from the fruit hull of mangosteen (Garcinia mangostana Linn.).

OBJECTIVE: The aim of this study was to investigate the antioxidative and neuroprotective activities of various extracts from the fruit hull of mangosteen (Garcinia mangostana Linn., GM). MATERIALS AND METHODS: Four extracts: water, 50% ethanol, 95% ethanol and ethyl acetate, were used. The antioxidative activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl free-radical scavenging assay at extract concentrations of 1, 10, 50 and 100 microg/ml. Based on the free radical scavenging activity of the extracts, two (water and 50% ethanol) were selected for their protective activity in NG108-15 neuroblastoma cells against H(2)O(2)-induced oxidative stress and for cell viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. RESULTS: All extracts exhibited antioxidative activity. The water and 50% ethanol extracts showed high free-radical scavenging activity with IC(50) values of 34.98 +/- 2.24 and 30.76 +/- 1.66 microg/ml, respectively. Both water and 50% ethanol extracts exhibited neuroprotective activity on NG108-15 cells. The highest activity was observed at the concentration of 50 microg/ml for both the water and 50% ethanol extracts. For cytotoxicity test, none of the extracts was toxic to the cells except at the high concentration of 100 microg/ml. CONCLUSIONS: These results suggest that the water and 50% ethanol extracts from the fruit hull of GM may be potent neuroprotectants.

Constituents of the pericarp of Glyptopetalum sclerocarpum.

Two new compounds, 6-oxo-20-hydroxy-20-epi-tingenol (1) and 2R*,3R*,5S*-trihydroxy-6R*-nonadecyltetrahydropyran-4-one (2), were isolated from the pericarp of Glyptopetalum sclerocarpum along with other 10 constituents.

Cytotoxic alkaloids from the flowers of Senna spectabilis.

Three new alkaloids, 3(R)-benzoyloxy-2(R)-methyl-6(R)-(11'-oxododecyl)-piperidine (3), 5-hydroxy-2-methyl-6-(11'-oxododecyl)-pyridine (4) and 5-hydroxy-2-methyl-6-(11'-oxododecyl)-pyridine N-oxide (5), together with a known alkaloid, (-)-cassine (1), were isolated from the flowers of Senna spectabilis. A derivative, N,O-diacetylcassine (2), was semisynthesized. Their structures and stereochemistry were established on the basis of spectroscopic analysis. Cytotoxic activity and brine shrimp lethality of these compounds were evaluated. Compounds 2, 3 and 5 exhibited cytoxicity against KB cell lines with IC50 values of 5.2, 3.7 and 2.0 microg/mL, respectively.

Quinone-methide triterpenoids from Glyptopetalum sclerocarpum.

Nine novel quinone-methide triterpenoids: 20-hydroxytingenone, 20,22beta-dihydroxytingenone, 20,22beta-dihydroxy-20-epi-tingenone, 20,21alpha-dihydroxy-22-oxo-21-desoxotingenone, 20-hydroxy-22-oxotingenone, 20-hydroxy-22-oxo-20-epi-tingenone, 21alpha-hydroxy-20,22-dioxo-30(20-->21)abeo-21-desoxotingenone, 20-oxo-20,21-seco-tingen-21-oic acid and 20-oxo-21-nor-20,21-seco-tingen-22-al, were isolated from the stem bark of Glyptopetalum sclerocarpum. Their structures were determined on the basis of spectroscopic evidence.