Identification of Apigenin and Luteolin in Artemisia annua L. for the Quality Control

Objective: To identify active compounds and establish the chemical fingerprint of Artemisia annua L. for the quality control. Methods: Thin-layer chromatography (TLC) conditions were developed to screen for 2 common flavonoids (apigenin and luteolin). Three mobile phases were used to isolate these flavonoids in 80% ethanolic extract of A. annua.Hexane : ethyl acetate : acetic acid (31:14:5, v/v) and toluene : 1,4-dioxane : acetic acid (90:25:4, v/v) were used in normal phase TLC (NP-TLC), and 5.5% formic acid in water : methanol (50:50, v/v) were used in reverse phase TLC (RP-TLC). Chromatograms were visualized under visible light after spraying with Fast Blue B Salt. Apigenin and luteolin bands were checked by comparing their Rf values and UV-Vis absorption spectra with reference markers. Results: Apigenin and luteolin were simultaneously detected with good specificity in RP-TLC condition, while only apigenin was detected in NP-TLC condition. Apigenin band intensity was higher than luteolin band intensity in both conditions. Conclusion: This knowledge can be applied to the development of quality control assessments to ensure product efficacy and consistenc

Identification of Apigenin and Luteolin in Artemisia annua L. for the Quality Control

Objective: To identify active compounds and establish the chemical fingerprint of Artemisia annua L. for the quality control. Methods: Thin-layer chromatography (TLC) conditions were developed to screen for 2 common flavonoids (apigenin and luteolin). Three mobile phases were used to isolate these flavonoids in 80% ethanolic extract of A. annua. Hexane : ethyl acetate : acetic acid (31:14:5, v/v) and toluene : 1,4-dioxane : acetic acid (90:25:4, v/v) were used in normal phase TLC (NP-TLC), and 5.5% formic acid in water : methanol (50:50, v/v) were used in reverse phase TLC (RP-TLC). Chromatograms were visualized under visible light after spraying with Fast Blue B Salt. Apigenin and luteolin bands were checked by comparing their Rf values and UV-Vis absorption spectra with reference markers. Results: Apigenin and luteolin were simultaneously detected with good specificity in RP-TLC condition, while only apigenin was detected in NP-TLC condition. Apigenin band intensity was higher than luteolin band intensity in both conditions. Conclusion: This knowledge can be applied to the development of quality control assessments to ensure product efficacy and consistency

The Effects of Thai Herbal Ha-Rak Formula on COX Isoform Expression in Human Umbilical Vein Endothelial Cells Induced by IL-1 β

Objective. To investigate the modulated effects of HRF on cyclooxygenase isoform expression and its activity, using the human umbilical vein endothelial cell (HUVEC) model induced by interleukin-1 beta (IL-1β). Methods. Cells were treated with indomethacin (positive control), HRF, and its components at various concentrations prior to treatment with IL-1β at 24 h. Cell viability was determined by MTT assay. Moreover, the anti-inflammatory effects of HRF and its components through mRNA and protein expression were established using real-time quantitative PCR and Western blot, respectively. COX activity was identified via exogenous and endogenous PGE2 productions using the EIA. Result. There was no cytotoxicity in HUVECs treated with HRF. None of the experimental conditions used in the study affected the expression of COX-1, but COX-2 protein expression was inhibited at concentrations under 10 µg/mL. Despite the significantly increased levels of exogenous PGE2, HRF had no effect on COX-2 mRNA expression. However, the production of PGE2 was lower at a concentration of 100 µg/mL HRF than at a concentration below 10 µg/mL. Interestingly, each component of HRF revealed different effects of the Ha-Rak formula. Conclusion. Our preliminary findings suggest that HRF and its components provide diverse modulation of COX-2 and PGE2 at the in vitro level

Additional file 1: of IL-1β-induced modulation of gene expression profile in human dermal fibroblasts: the effects of Thai herbal Sahatsatara formula, piperine and gallic acid possessing antioxidant properties

The overview of microarray analysis work flow (Figure S1) and the list of human housekeeping genes matching with gene array in microarray BeadChip (Table S1). (DOCX 32 kb

Effect of Thunbergia laurifolia Herbal Tea on Glucose Homeostasis in Healthy Volunteers: A Single-Arm Phase I Study

Background. Thunbergia laurifolia (TL) is a commonly used herbal medicine in Thailand and in other Asian countries. TL has been approved as a Thai traditional medicine for detoxifying poisons, and the list of possible adverse effects includes hypoglycemia. TL showed hypoglycemic effect in animals possibly due to antioxidant effect and beta-cell preservation. However, the safety of TL herbal tea and its effects on glucose homeostasis have never been investigated in humans. Methods. Twenty healthy volunteers (10 men and 10 women) drank TL herbal tea 3 times/day for 2 weeks. Ten subjects took TL herbal tea 9 grams daily. After the safety of TL herbal tea was established, 10 more subjects took TL 12 grams daily. Clinical and biochemical tests were assessed at baseline and at 2 weeks. Results. Mean age was 34.9 ± 10.2 years, and mean body mass index was 27.5 ± 5.8 kg/m2. Baseline and posttreatment plasma concentrations were as follows: fasting plasma glucose (89 ± 6 vs. 89 ± 7 mg/dL), fructosamine (213 ± 32 vs. 212 ± 33 μmol/L), fasting insulin (8.8 [IQR: 5.9–18.4] vs. 10.4 [IQR: 7.4–15.2] μU/mL), HOMA-B (101.6 [IQR: 82.3–189.8] vs. 120.4 [IQR: 93.2–153.2]), and HOMA-IR (1.1 [IQR: 0.8–2.3] vs. 1.4 [IQR: 0.9–2.0]), all respectively. There were no significant changes in these parameters, including body weight, blood pressure, lipid profile, and C-reactive protein. No serious adverse events were observed during the study period. Conclusions. TL herbal tea at doses of 9 and 12 grams daily had good tolerability without any significant adverse effects on fasting plasma glucose level or other glucose homeostasis parameters measured

Chemical Profiling of an Antipyretic Drug, Thai Herbal Harak Formula, by Liquid Chromatography Coupled with Quadrupole Time-of-Flight Mass Spectrometry

Objective: To isolate and identify chemical compounds in Harak, a Thai herbal formula widely used as an antipyretic drug in Thailand. Methods: Methanol extraction of the Harak formula and its five herbal components were separated by Ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. This method has been developed for “untargeted” profiling of this formula and its components. An online database was used to identify chemical compounds by comparing its empirical molecular formula, isotope pattern, and fragmentation pattern. Results: Nineteen chemical constituents were found from positive and negative electrospray ionization (ESI) modes. These compounds included flavonoids, hesperitine, and iso-corydine, which are known to possess antioxidant and anticancer activity. Moreover, data from the principle component analysis (PCA) score plot of positive and negative ESI modes showed that the chemical constituents of Thai herbal Harak formula were similar to those found in Ficus racemosa Linn. and Capparis micracantha DC. Conclusion: Under this optimization method, nineteen chemical constituents including phenolic and flavonoids were characterized in both positive and negative ESI mode

The Polyherbal Wattana Formula Displays Anti-Amyloidogenic Properties by Increasing α-Secretase Activities.

Alzheimer's disease is characterized by the deposition of insoluble amyloid-β peptides produced from the β-amyloid precursor protein (βAPP). Because α-secretase cleavage by ADAM10 and ADAM17 takes place in the middle of Aβ, its activation is considered as a promising anti-AD therapeutic track. Here we establish that the polyherbal Wattana formula (WNF) stimulates sAPPα production in cells of neuronal and non-neuronal origins through an increase of both ADAM10 and ADAM17 catalytic activities with no modification of BACE1 activity and expression. This effect is blocked by specific inhibition or genetic depletion of these disintegrins and we show that WNF up-regulates ADAM10 transcription and ADAM17 maturation. In addition, WNF reduces Aβ40 and Aβ42 generation in human cell lines. Altogether, WNF presents all the characteristics of a potent preventive anti-Alzheimer formula. Importantly, this natural recipe, currently prescribed to patients for the treatment of other symptoms without any secondary effect, can be tested immediately for further clinical studies

Metabolomics and Integrative Omics for the Development of Thai Traditional Medicine

In recent years, interest in studies of traditional medicine in Asian and African countries has gradually increased due to its potential to complement modern medicine. In this review, we provide an overview of Thai traditional medicine (TTM) current development, and ongoing research activities of TTM related to metabolomics. This review will also focus on three important elements of systems biology analysis of TTM including analytical techniques, statistical approaches and bioinformatics tools for handling and analyzing untargeted metabolomics data. The main objective of this data analysis is to gain a comprehensive understanding of the system wide effects that TTM has on individuals. Furthermore, potential applications of metabolomics and systems medicine in TTM will also be discussed

WNF up-regulates ADAM10 and ADAM17 via two distinct mechanisms in human HEK293 cells.

<p>(A, B) HEK293 cells were treated for 36 hours without (control) or with WNF (100μg/ml) in DMEM/1% FBS. (A) ADAM10 (left panel), ADAM17 (right panel) as well as β-actin immunoreactivities were assayed by western blot. (B) ADAM10, ADAM17 as well as GAPDH mRNA levels were determined by real-time PCR. Black bars in histograms correspond to the densitometric analyses normalized with β-actin (A) or mRNA levels normalized with GAPDH (B), are expressed as a percentage of control (non-treated cells, white bars) and represent the means ± SE of 11 to 17 independent determinations. *p<0.0005; **p<0.0001; ns, non-statistically different. (C) HEK293 were incubated for 16 hours without (control) or with WNF (100μg/ml), homogenized and the sub-cellular distributions of endogenous ADAM10 (panels a and b), ADAM17 (panels c and d) as well as the ER marker calnexin (panel e) and the Golgi marker Golgi 58K protein (panel f) were analyzed by western blot after sucrose gradient fractionation. TGN, trans-golgi network; ER/PM, endoplasmic reticulum/plasma membrane.</p