A double-blinded, randomized, split-side, vehicle-controlled study of the efficacy of cleanser containing Acanthus ebracteatus Vahl., Suregada multiflora, and Acacia concinna in patients with atopic dermatitis: A pilot study.

BACKGROUND: Barrier repair therapy is the key management approach for both eczematous and non-lesional skin of atopic dermatitis. The use of appropriate cleansers to enhance skin hydration is an adjunctive treatment that increases topical drug penetration. Anti-inflammatory properties of various medicinal plants in tropical Asia have been reported. OBJECTIVE: Investigate the efficacy of herbal cleanser containing a combination of herbal extracts from Acanthus ebracteatus Vahl., Suregada multiflora, and Acacia concinna on seemingly intact skin in patients with atopic dermatitis by measuring improvements in the skin barrier function. METHODS: This 2-week pilot study was a split-side, randomized, double-blinded, vehicle-controlled trial. All patients (n = 30) were asked to use both a cleanser with an active formulation containing the herbal extracts and a vehicle- controlled cleanser on each side of mid-volar forearm. Biophysical assessments including transepidermal water loss (TEWL), skin hydration, skin pH, and skin roughness were performed at baseline and upon study completion. RESULTS: Compared to baseline, the median percentage change in TEWL at the end of the study was significantly greater for the active side 10.4 (-19, 20.7) g/m2h than the control side -13.2 (-28.7, 9.1) g/m2h; p = 0.01. The median percentage change of skin hydration, skin pH, and skin roughness of the active side compared to the control side had no a statistical significance. CONCLUSIONS: This cleanser is beneficial when used as adjunctive therapy. Further studies should evaluate its anti- sinflammatory properties in the remedy or active phase of atopic dermatitis or other inflammatory skin diseases.

Thai herbal antipyretic 22 formula (APF22) inhibits UVA-mediated melanogenesis through activation of Nrf2-regulated antioxidant defense.

Thai herbal antipyretic 22 formula (APF22), a polyherbal formula, has been traditionally used to treat dermatologic problems including hyperpigmentation. Exposure of the skin to ultraviolet A (UVA) causes abnormal melanin production induced by photooxidative stress. This study thus aimed to investigate the protective effects of APF22 extracts and phenolic compounds, ferulic acid (FA), and gallic acid (GA; used as positive control and reference compounds), on melanogenesis through modulation of nuclear factor E2-related factor 2 (Nrf2) signaling and antioxidant defenses in mouse melanoma (B16F10) cells exposed to UVA. Our results revealed that the APF22 extracts, FA, and GA reduced melanin synthesis as well as activity and protein levels of tyrosinase in UVA-irradiated B16F10 cells. Moreover, APF22 extracts and both FA and GA were able to activate Nrf2-antioxidant response element signaling and promote antioxidant defenses including glutathione, catalase, glutathione peroxidase, and the glutathione-S-transferase at both mRNA and enzyme activity levels in irradiated cells. In conclusion, APF22 extracts suppressed UVA-mediated melanogenesis in B16F10 cells possibly via redox mechanisms involving activation of Nrf2 signaling and upregulation of antioxidant defenses. Moreover, pharmacological action of the APF22 extracts may be attributed to the phenolic compounds, FA, and GA, probably serving as the APF22's active compounds.

Activation of Nrf2 Reduces UVA-Mediated MMP-1 Upregulation via MAPK/AP-1 Signaling Cascades: The Photoprotective Effects of Sulforaphane and Hispidulin.

UVA irradiation plays a role in premature aging of the skin through triggering oxidative stress-associated stimulation of matrix metalloproteinase-1 (MMP-1) responsible for collagen degradation, a hallmark of photoaged skin. Compounds that can activate nuclear factor E2-related factor 2 (Nrf2), a transcription factor regulating antioxidant gene expression, should therefore serve as effective antiphotoaging agents. We investigated whether genetic silencing of Nrf2 could relieve UVA-mediated MMP-1 upregulation via activation of mitogen-activated protein kinase (MAPK)/activator protein 1 (AP-1) signaling using human keratinocyte cell line (HaCaT). Antiphotoaging effects of hispidulin (HPD) and sulforaphane (SFN) were assessed on their abilities to activate Nrf2 in controlling MMP-1 and collagen expressions in association with phosphorylation of MAPKs (extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38), c-Jun, and c-Fos, using the skin of BALB/c mice subjected to repetitive UVA irradiation. Our findings suggested that depletion of Nrf2 promoted both mRNA expression and activity of MMP-1 in the UVA-irradiated HaCaT cells. Treatment of Nrf2 knocked-down HaCaT cells with MAPK inhibitors significantly suppressed UVA-induced MMP-1 and AP-1 activities. Moreover, pretreatment of the mouse skin with HPD and SFN, which could activate Nrf2, provided protective effects against UVA-mediated MMP-1 induction and collagen depletion in correlation with the decreased levels of phosphorylated MAPKs, c-Jun, and c-Fos in the mouse skin. In conclusion, Nrf2 could influence UVA-mediated MMP-1 upregulation through the MAPK/AP-1 signaling cascades. HPD and SFN may therefore represent promising antiphotoaging candidates.

13C-Acetic Acid Breath Test Monitoring of Gastric Emptying during Disease Progression in Diabetic Rats.

Gastric motility disturbance is commonly found in long-standing hyperglycemia. Both delayed and rapid gastric emptying has been reported in diabetes. However, very few studies have followed the changes in gastric emptying during disease progression in diabetes because of technical limitations. 13C-Acetic acid breath test is a validated method which is non-invasive and can be used repeatedly or serially to evaluate gastric emptying changes in animal. We investigated the gastric emptying changes in different stages of diabetes using 13C-acetic acid breath test, as well as its related mechanisms involving interstitial cells of Cajal (ICCs), and stem cell factor (SCF) in streptozotocin-induced diabetic rats. The results showed that gastric emptying was accelerated at the early stage (12 weeks of diabetes) whereas intramuscular ICCs (ICC-IM) networks were not different from normal group. At long-term stage (28 weeks of diabetes), gastric emptying had returned to normal pattern with no delayed. ICC-IM networks were decreased in the diabetic group compared to 12th weeks, and were lower than in the normal group at the same time point. SCF levels were constantly high in the diabetic group than in the normal group. This result indicated that 13C-acetic acid breath test is useful to track the alteration in gastric emptying during disease progression. The change of gastric emptying was not found to be significantly associated with ICC-IM. Elevated SCF may help to preserve ICC-IM, especially in the early phase of diabetes.

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

BACKGROUND: Pain is the main symptom of most musculoskeletal disorders and can be caused by inflammation in association with oxidative stress. Thai herbal Sahatsatara formula (STF), a polyherbal formula, has been traditionally used for relieving muscle pain and limb numbness. This study aimed to investigate biologically active compounds of STF and its pharmacological effects related to antioxidant and anti-inflammatory activities. METHODS: The identification of possibly active compounds of STF was performed by high performance liquid chromatography (HPLC). Moreover, this study also assessed the free radical scavenging activities of STF and its components using DPPH radical scavenging assay and their inhibitory effects on IL-1ß-induced intracellular reactive oxygen species (ROS) formation in primary human dermal fibroblasts (NHDFs) using DCFDA-flow cytometry analysis. Modulation of human gene expression by STF and its active compounds was investigated by microarray analyzed through Gene Ontology (GO) classification and pathway enrichment analysis. RESULTS: HPLC analysis has revealed the presence of gallic acid (GA) and piperine (PP) as the major compounds in STF extracts. Our finding discovered that STF and its active compounds (GA and PP) yielded free radical scavenging activities and abilities to inhibit IL-1ß-induced cellular ROS formation in NHDFs. Furthermore, microarray analysis demonstrated that a total of 84 genes (54 upregulated and 30 downregulated) were significantly affected by IL-1ß involved in inflammatory cytokines, chemokines, transcription factors, cell adhesion molecules and other immunomodulators participating in NF-κB signaling. The significantly upregulated genes in IL-1ß-treated in NHDFs participate in interleukin and cholecystokinin (CCRK) signaling pathways. The GO analysis of the target genes showed that all test compounds including indomethacin, STF and its active compounds, can downregulate the genes involved in NF-кB signaling pathway in IL-1ß-treated NHDFs compared to the cells treated with IL-1ß alone. CONCLUSIONS: STF and its active compounds possessing antioxidant actions can modulate the effects of IL-1ß-mediated alteration of gene expression profiles associated with inflammatory signaling in NHDFs.

The effects of the court-type Thai traditional massage on anatomical relations, blood flow, and skin temperature of the neck, shoulder, and arm.

BACKGROUND: Court-type Thai traditional massage (CTTM) has specific major signal points (MaSP) for treating musculoskeletal conditions. The objectives of this study are to investigate the anatomical surfaces and structures of MaSPs, and to examine blood flow (BF) and skin temperature (ST) changes after applying pressure on the MaSPs on neck, shoulder, and arm areas. METHODS: In the anatomical study, 83 cadavers were dissected and the anatomical surfaces and structures of the 15 MaSPs recorded. In human volunteers, BF, peak systolic velocity (PS), diameter of artery (DA), and ST changes were measured at baseline and after pressure application at 0, 30, 60, 180, and 300 s. RESULTS: There was no statistical difference in anatomical surfaces and structures of MaSP between the left and right side of the body. The 3 MaSPs on the neck were shown to be anatomically separated from the location of the common carotid arteries. The BF of MaSPs of the neck significantly and immediately increased after pressure application for 30 s and for 60 s in the arm (p < 0.001). ST increased significantly and immediately after pressure application for 300 s (p < 0.001). There was no significant correlation between BF and ST at any of the MaSPs. CONCLUSIONS: This study showed that MaSP massages were mainly directed towards muscles. MaSPs can cause significant, but brief, increases in BF and ST. Further studies are suggested to identify changes in BF and ST for all of the MaSPs after actual massage treatment sessions as well as other physiological effects of massage.

Effects of the Ayurved Siriraj Wattana recipe on functional and phenotypic characterization of cytokine-induced killer cells and dendritic cells in vitro.

BACKGROUND: Ayurved Siriraj Wattana recipe (AVS073), has been prescribed as tonic, to increase appetite, and for pain relief. It also exhibits antioxidant, anti-inflammatory, immunomodulating and anti-cancer activities. However, the immunomodulatory effects on antigen-presenting cells and effector T cells remained elusive. We thus aimed to study the effects of AVS073 on differentiation, maturation, functions and proportions of CIK cells and monocyte-derived DCs. METHODS: CIK cells and monocyte-derived DCs were treated with AVS073, followed by the assessment of T-helper (Th) phenotypes using real-time RT-PCR and flow cytometry. RESULTS: AVS073 promoted Th1 phenotype in CD3+CD56+ subset of CIK cells through increasing STAT4, T-bet, and interferon-γ. AVS073 inhibited Th2 phenotype through decreasing STAT6. AVS073 inhibited Treg phenotype through decreasing STAT5A, STAT5B and IDO. AVS073 promoted Th17 phenotype through increasing STAT3, RORC and IL-17. AVS073 treatment of mDCs resulted in increasing Th1-prone cytokine (IL-12) and Th17-prone cytokines (IL-6 and IL-23). CONCLUSIONS: AVS073 upregulated Th1 and Th17, but downregulated Th2 and Treg phenotypes within CD3+CD56+ cells. The treatment of mDCs drove Th1 and Th17-polarizations.

Moringa Oleifera Leaf Increases Insulin Secretion after Single Dose Administration: A Preliminary Study in Healthy Subjects.

BACKGROUND: Herbal medicine has long been used as an alternative medicine for treatment of type 2 diabetes mellitus (T2DM). Recently, Moringa oleifera (MO or ma-rum in Thai) leaf has been widely used in T2DM patients. Several studies in diabetes rat model have shown that MO had effect on glucose metabolism. However study in humans is lacking. OBJECTIVE: Examine effects of MO on plasma glucose and insulin secretion. MATERIAL AND METHOD: Ten healthy volunteers were enrolled in this study (mean age 29 ± 5 years; BMI 20.6 ± 1.5 kg/m2; FPG 81 ± 5 mg/dl). After an overnight fast and every two weeks, subjects received an oral dose of MO at increasing dosages of 0, 1, 2, and 4 g. Plasma glucose (PG) and insulin were collected at baseline and at 0.5, 1, 1.5, 2, 4, and 6 hours after each MO dosage administration. Insulin secretion rate was measured using area under the curve (AUC) of insulin and AUC of insulin/glucose ratio. RESULTS: After doses of 0, 1, 2, and 4 g MO, mean plasma insulin increased (2.3 ± 0.9, 2.7 ± 1.0, 3.3 ± 1.4, and 4.1 ± 1.7 µU/ml, respectively) despite there being no differences in mean PG (77 ± 6, 78 ± 5, 79 ± 6, and 79 ± 5 mg/dl, respectively). AUC of insulin was greater after high-dose MO (4 g) than after baseline or low-dose MO capsule (1 g) (24.0 ± 3.5 vs. 14.5 ± 1.8 or 16.1 ± 2.0, respectively; p = 0.03), while there was no difference in AUC of glucose. Accordingly, insulin secretion rate represented by AUC of insulin/glucose ratio after high-dose MO was significantly increased by 74% (P = 0.041), as compared with that of baseline. CONCLUSION: We concluded that high-dose (4 g) MO leaf powder capsules significantly increased insulin secretion in healthy subjects. These results suggest that MO leaf may be a potential agent in the treatment of type 2 diabetes. Further studies of MO in patients with T2DM are needed.

Therapeutic potential of Thai herbal formula for cognitive impairment: A metabolomics approach for Comprehensive Insights.

Chronic cerebral ischemia hypoperfusion plays a role in the initiation and progression of vascular dementia, which causes changes in metabolites. Currently, there is no standard treatment to treat, prevent and reduce the severity of this condition. Thai herbal Yahom no.20 (YHF20) is indicated for fatigue and dizziness. The components of YHF20 have been found to have pharmacological effects related to the pathology of chronic cerebral ischemia hypoperfusion. This study aimed to investigate metabolomic changes after YHF20 administration in a rat model of permanent bilateral common carotid artery occlusion (2-VO) induced chronic cerebral ischemia hypoperfusion, and to explore its impact on spatial learning and memory. Albino Wistar rats were randomly allocated to 5 groups; sham, 2-VO, 2-VO+ 100 mg/kg YHF20, 2-VO+300 mg/kg YHF20, and 2-VO+1000 mg/kg YHF20. The rats were administered YHF20 daily by oral gavage for 56 days after 2-VO induction. Plasma was collected weekly for metabolome change analysis using LC-MS/QTof and toxicity study. The rats were evaluated for spatial learning and memory using the Morris water maze. The results showed that 78 known metabolites and 10 tentative pathways altered after chronic cerebral hypoperfusion, although it was not able to determine the effect on memory and learning behaviors of rats. Glutathione and glutathione metabolism might be metabolite-pathway that were the affect after YHF20 administration in cerebral ischemic condition. The 4 known metabolites may be the metabolites from the constituents of YHF20 could be considered and confirmed for quality control purpose. In conclusion, YHF20 administration might contribute to metabolic changes related to cerebral ischemia condition without the effect on spatial learning and memory, including hepatotoxicity and nephrotoxicity after 56 days of treatment. Alterations in the potential metabolites may provide data support for elucidating dementia pathogenesis and selecting pathways for intervention.

Protective effect of AVS073, a polyherbal formula, against UVA-induced melanogenesis through a redox mechanism involving glutathione-related antioxidant defense.

BACKGROUND: Ayurved Siriraj Brand Wattana formula (AVS073), a Thai herbal formula, has traditionally been used for health promotion and prevention of age-related problems. Ultraviolet A (UVA) is recognized to play a vital role in stimulation of melanin synthesis responsible for abnormal skin pigmentation possibly mediated by photooxidative stress. We thus aimed to study the inhibitory effect of AVS073 extracts on UVA-induced melanogenesis via a redox mechanism involving glutathione (GSH) synthesis and glutathione S-transferase (GST) using human melanoma (G361) cell culture. METHODS: The standardization of AVS073 extracts was carried out by TLC and UHPLC to obtain fingerprinting profiles of the formula, which identified several phenolic compounds including gallic acid (GA) in the formula. Antimelanogenic actions of AVS073 (up to 60 µg/ml) and GA (up to 10 µg/ml) were investigated by measuring tyrosinase activity and mRNA as well as melanin level in G361 cells irradiated with UVA. Moreover, antioxidant actions of the herbal formula and GA were determined by evaluating oxidant formation and modulation of GSH-related antioxidant defenses including GSH content, GST activity and mRNA level of γ-glutamate cysteine ligase catalytic (γ-GCLC) and modifier (γ-GCLM) subunit and GST. RESULTS: AVS073 extracts and GA, used as a reference compound, suppressed UVA-augmented tyrosinase activity and mRNA and melanin formation. In addition, pretreatment with AVS073 and GA was able to inhibit cellular oxidative stress, GSH depletion, GST inactivation and downregulation of γ-GCLC, γ-GCLM and GST mRNA in G361 cells exposed to UVA radiation. CONCLUSIONS: AVS073 formula exerted antimelanogenic effects possibly through improving the redox state by upregulation of GSH and GST. Moreover, pharmacological activity of the polyherbal formula would be attributed to combined action of different phenolic compounds present in the formula.

A comparative study of chemical constituents and safety of Thai herbal medicated oil formula and traditional medicated oil.

OBJECTIVES: Medicated oil has been traditionally used as an analgesic medicine to relieve pain by applying to the skin over painful muscles, and has been established in The National List of Essential Medicines (2556 B.E.). Traditional Medicated oil (TMO) consists of cassumunar ginger, Wan En Lueang, and Wan Ron Thong. It is prepared by frying herbs in coconut oil which is a traditional method. Thai Herbal Medicated oil formula (MOF) comprises cassumunar ginger, sweet flag, lesser galangal, wild turmeric, Wan En Lueang, and Wan Ron Thong. Its preparation by maceration with methyl salicylate is modified method to avoid the heat in the traditional preparation. Since both recipes have different methods of extraction and compositions, it is necessary to determine the chemical constituents and acute skin irritation potential of MOF and TMO. METHODS: This study applied Gas Chromatography-Mass Spectrometry (GC-MS) for quality assessment of MOF extract and TMO extract. The mass spectra of the compounds matched with authentic standards from the NIST library. The compounds were identified by comparing their retention time and peak area, and the percentage of matched factor was more than 80%. The safety assessment on acute skin irritation used the 4 h human patch test (4 h HPT) in 36 healthy subjects. RESULTS: The 37 compounds were found in MOF extract. The most common chemical constituents were terpene derivatives; monoterpenes and sesquiterpenes, and others, namely benzene derivatives, ester, phenylpropanoid, and other compounds. The 33 compounds were found in TMO extract and the chemical groups were similar to MOF extract. The 4 h HPT demonstrated that the MOF extract, TMO extract, methyl salicylate solvent, and coconut oil solvent, did not induce skin irritation. However, the vehicle of the formulae and the whole formulae as MOF and TMO clinically induced skin irritation and accorded with rubefacient. The characteristic as mild erythema or dryness, but not erythema with edema, could appear after the use of rubbing skin products. CONCLUSIONS: The chemical constituents found in MOF and TMO extracts were partially different and their acute skin irritation reactions were not significantly different. A study on the efficacy of both remedies should be performed in the future, as well as a study on active chemical constituents. This study provides scientific evidences of quality and safety, including GC-MS condition for poly-herbal medicated oil standardization.

Metabolomic analysis of Thai Herbal Analgesic Formula based on ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.

Sahatsatara formula (STF), a Thai herbal medicine formula which has been commonly used as analgesic drugs to relieve musculoskeletal pain and numbness in Thai traditional medicine. The pharmacological studies of its ingredients have represented that have anti-inflammatory and antioxidant properties. However, the quality markers (Q-markers) for STF are still unknown and require further investigation. The primary goal of this study was to establish the chemical profile of STF though metabolomic analysis. Untargeted metabolomics were used to analyze global components and accurately qualify compounds. Multivariate analysis (MVA) was used to classify STF extract at three different concentrations and a quality control sample. Furthermore, samples' characteristics and identification-related markers were observed and compounds matched to the Traditional Chinese medicine library in UNIFI software. According to the results, chemical analysis revealed 63 compounds in positive mode and 33 compounds in negative mode within STF. Notably, 19 potential Q-markers were tentative identified in all three concentrations of STF, including alkaloids, terpenes, phenols, organic acids, disaccharides, fatty acids, glycosides, quinonoids, and steroids. The compounds exhibited pharmacological effects such as anti-inflammatory activity, anti-oxidant activity, and analgesic properties, which correlated to traditional properties of STF. Consequently, this study provides insights into the chemical profiles of the STF and identifies potential markers that can be utilized for qualitative and quantitative quality control of STF. Additionally, the findings can also be useful for further research into STF's anti-inflammatory properties through in vitro assays, as well as exploring its clinical efficacy to support evidence-based medicine for STF.

Metabolomics of Personalized Body Elements in Thai Traditional Medicine Response to Herbal Medicine for Body Elements Balancing in Healthy Volunteers.

Background: In Thai traditional medicine (TTM), the dominant body element called "Dhat Chao Ruean" (DCR) is an integral part in the diagnostic process of Thai traditional medicine. TTM practitioners usually use Thai herbal Benjakul formula (BKF) for adjusting and balancing the body elements. However, the effects of BKF on metabolism and individual response to it have not been studied yet. Methods: This study proposed to investigate the metabolic profiling in 24 volunteers categorized by their types of birth month DCR (bDCR) after the administration of BKF (450 mg, three tablets three times a day before meals) for seven days. Differences in metabolic profiling between bDCR groups were investigated by using liquid chromatography coupled with mass spectrometry for untargeted analysis, and in addition, the safety was assessed by testing the plasma biochemical level. Results: This study identified 57 biomarkers in positive ESI and 12 in negative ESI. Piperine was found in varying amount among the participants but it was the highest in the earth group. In addition, this study found that elemicin, phenylpropionic acid, ricinoleic acid, and ß-sitosterol are important substances in a single herb of BKF. Regarding biochemical tests, the results indicated that BKF can decrease the lipid profile and it has no toxic effects on liver and kidney functions. Conclusion: The findings indicated that it is safe to use BKF which can help to improve health in chronic diseases by adjusting abnormality of the elements of the body. In addition, the information gathered from this study is valuable for further study in the field of Thai traditional medicine.

Prolonged Egg Supplement Advances Growing Child's Growth and Gut Microbiota.

Protein-energy malnutrition still impacts children's growth and development. We investigated the prolonged effects of egg supplementation on growth and microbiota in primary school children. For this study, 8-14-year-old students (51.5% F) in six rural schools in Thailand were randomly assigned into three groups: (1) whole egg (WE), consuming 10 additional eggs/week (n = 238) (n = 238); (2) protein substitute (PS), consuming yolk-free egg substitutes equivalent to 10 eggs/week (n = 200); and (3) control group (C, (n = 197)). The outcomes were measured at week 0, 14, and 35. At the baseline, 17% of the students were underweight, 18% were stunted, and 13% were wasted. At week 35, compared to the C group the weight and height difference increased significantly in the WE group (3.6 ± 23.5 kg, p < 0.001; 5.1 ± 23.2 cm, p < 0.001). No significant differences in weight or height were observed between the PS and C groups. Significant decreases in atherogenic lipoproteins were observed in the WE, but not in PS group. HDL-cholesterol tended to increase in the WE group (0.02 ± 0.59 mmol/L, ns). The bacterial diversity was similar among the groups. The relative abundance of Bifidobacterium increased by 1.28-fold in the WE group compared to the baseline and differential abundance analysis which indicated that Lachnospira increased and Varibaculum decreased significantly. In conclusion, prolonged whole egg supplementation is an effective intervention to improve growth, nutritional biomarkers, and gut microbiota with unaltered adverse effects on blood lipoproteins.

The safety of Homnawakod herbal formula containing Aristolochia tagala Cham. in Wistar rats.

BACKGROUND: A dried root of Aristolochia tagala Cham. (ATC) is often used in Thai traditional medicine as an antipyretic, anti-inflammatory agent, muscle relaxant, appetite-enhancing agent, and analeptic. Homnawakod, an important herbal recipe, originally contains ATC in its formula, however, some Aristolochia species have been reported to cause nephrotoxicity due to aristolochic acid (AA) and its derivatives, resulting in ATC removal from all formulae. Therefore, this study investigates the chemical profiles of ATC, the original (HNK+ATC) and the present Homnawakod Ayurved Siriraj Herbal Formulary™ (HNK), and investigates whether they could cause nephrotoxicity or aggravate LPS-induced organ injuries in vivo. METHODS: HPLC and LC/MS were used for chemical profile study. Male Wistar rats were randomly divided into groups in which the rats were intragastrically administered distilled water (2 groups), ATC (10 or 30 mg/kg), HNK+ATC (540 or 1,620 mg/kg), or HNK (1,590 mg/kg) for 21 days. A positive control group was administered with single dose 100 mg/kg standard AA-I intragastrically at day 1. Serum creatinine and urea were measured at baseline and at 7, 14 and 21 days of the treatment. On day 22, a model of lipopolysaccharide (LPS)-induced endotoxemia was used. One-way and two-way analyses of variance were performed and a P value of less than 0.05 was considered to be significant. RESULTS: The similarity of the HPLC chromatograms of HNK+ATC and HNK could suggest that the qualities of both formulae are nearly the same in terms of chemical profile. The amount of AA-I found in ATC is 0.24%w/w. All experimental groups exhibited similar levels of serum urea at baseline and 7 and 14 days of the treatment. At 21 days, rats received AA exhibited a significant increase in serum urea, whereas the others did not exhibit such toxicity. On day 22, there were no significant changes in LPS-induced renal and liver dysfunction, or LPS-induced mean arterial pressure (MAP) reduction upon administration of ATC, HNK+ATC, HNK or AA-I. CONCLUSIONS: These results suggest that ATC, HNK+ATC or HNK, at the animal dose equivalent to that used in human, do not cause the acute nephrotoxicity in rats and do not aggravate LPS-induced organ injuries even further.

Phase II clinical trial of Ayurved Siriraj Wattana Recipe for symptomatic relief in patients with osteoarthritis of the knee.

BACKGROUND: Osteoarthritis of the knee is one of the most common public health problems in Thailand and throughout the world. It causes disability, with pain and loss of function. Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most common treatment modality to relieve pain for OA, but can cause undesirable adverse effects. Effective and safer alternative treatments for OA are urgently needed. Among the many kinds, Ayurved Siriraj Wattana Recipe has been widely used in humans for a long time. The components of the recipe probably were related with many systems of the body such as the immune system and the gastrointestinal system and it may have anti-inflammatory activity as well as anti-oxidative capacity. OBJECTIVE: Determine the efficacy of Ayurved Siriraj Wattana Recipe for knee osteoarthritis in a randomized, open-label, diclofenac-controlled trial for 12 weeks. The trial has been approved from Siriraj Institutional Review Board (SIRB), Faculty of Medicine, Siriraj Hospital, Mahidol University. MATERIAL AND METHOD: Analyze the 12-item Oxford, defined as 5-point Likert scales of pain and functional activity. The secondary assessment included 10-cm VAS pain assessment tool as well as, patients' and physician's global efficacy assessment. RESULTS: "12-Item Oxford" revealed that patients had mild problems in terms of function and pain with an average function score observed between groups (p = 0.6). The average pain score at 12 weeks' follow-up was not significantly different in the Wattana group compared with the diclofenac group (p = 0.3). Responses with no differences included the mean 10-cm VAS that assessed pain at 12 weeks' follow-up in both groups. There were no significant differences in the patients' and physician's global efficacy assessment with patients in both groups. There were no differences in the frequent of adverse events between groups. In addition, there were no significant differences in the patients' and physician's global safety assessment between groups. CONCLUSION: The present, diclofenac-controlled trial concludes that Ayurved Siriraj Wattana Recipe treatment could be an effective treatment of pain in knee OA at 12 weeks.

The Phyllanthus emblica L. infusion carries immunostimulatory activity in a mouse model.

BACKGROUND: Phyllanthus emblica L. (Indian gooseberry, Ma khaam pom) has been an herbal component of Thai traditional recipes proposed to slow down the aging process. A number of methodologies have been employed to investigate the immunological aspects of the so called "anti-aging effects" of P. emblica in a BALB/c mice model. OBJECTIVE: 1) To investigate the immunological efficacy of the anti-aging effects of P. emblica infusion in a BALB/c mice model. 2) To verify the safety for the consumption of P. emblica infusion in BALB/c mice. MATERIAL AND METHOD: For in vitro studies, splenocytes were isolated from mice and examined in comparison with the human umbilical endothelial cells, fibroblasts and YAC-1 (mouse lymphoma) cells for proliferative activity upon the exposure to P. emblica infusion. For in vivo studies, mice were orally administered with P. emblica infusion at a dose range of 0, 50, 100, 200 mg/kg BW for 14 days. After the treatments, splenocytes isolated from these mice examined for proliferative and NK cell activities. RESULTS: For in vitro studies, the infusion of P. emblica could directly drive the proliferation of mouse splenocytes in a dose-dependent manner. The P. emblica infusion itself was already cytotoxic to YAC-1 in the studied dose, while sparing the human umbilical endothelial cells and fibroblasts. For in vivo studies, splenocytes isolated from these mice exhibited dose-dependent proliferative activities. Only the isolated splenocytes from mice ingesting 100 mg/kg BW exhibited an enhancement in NK cell activity. CONCLUSION: P. emblica infusion could drive proliferative activity of splenocyte in vitro and in vivo, with an enhancement in the NK cell-induced cytotoxic activity. The infusion in the aforementioned dose was safe throughout the study.

Utilization of UPLC-PDA and GC-MS/MS coupled with metabolomics analysis to identify bioactive metabolites in medicinal turmeric at different ages for the quality assurance.

BACKGROUND: Available monographs often suggest measurement of curcumin or curcuminoids for quality control of turmeric-based medicines/products. However, one compound is not enough to indicate the quality of traditional herbal medicines due to the holistic approach. Furthermore, to ensure high quality of such products, good harvesting practice plays an important role in the quality control of turmeric raw materials. PURPOSE: This study aimed to indicate quality markers of turmeric and to suggest optimum harvesting times for turmeric used for medicinal purposes by integration of Thai traditional medicine knowledge. METHODS: Turmeric rhizomes at 4, 6, 8, 9, 10, and 11 months old were analyzed. UPLC-PDA was used for quantitation of curcumin. GC-MS/MS was used to obtain chemical profiles of turmeric volatile oil. PCA, Volcano plot, and HCA were performed to identify similarities or differences of the data. RESULTS: Turmeric aged between 6 and 10 months old contained > 5%w/w of curcumin, which complied with Thai Herbal Pharmacopoeia 2021. GC-MS/MS analysis suggested suitable chemical markers, namely Ar-turmerone, turmerone, curlone, and zingiberene, because they exhibited pharmacological activities related to the traditional uses of turmeric. Eucalyptol, santalene, ß-caryophyllene, cis-ß-farnesene, α-caryophyllene, curcumene, ß-bisabolene, ß-sesquiphellandrene, and cis-sesquisabinene hydrate were also tentatively identified. Later, the multivariate analysis revealed that turmeric aged between 6 and 10 months old showed similar metabolite profiles. CONCLUSION: Based on curcumin content and chemical profiles, turmeric aged between 6 and 10 months old are appropriate for uses in Thai traditional medicine. Our study suggests additional information in the quality control of turmeric to assure its efficacy, especially for uses in traditional medicines.

Upregulation of CYP 450s expression of immortalized hepatocyte-like cells derived from mesenchymal stem cells by enzyme inducers.

BACKGROUND: The strenuous procurement of cultured human hepatocytes and their short lives have constrained the cell culture model of cytochrome P450 (CYP450) induction, xenobiotic biotransformation, and hepatotoxicity. The development of continuous non-tumorous cell line steadily containing hepatocyte phenotypes would substitute the primary hepatocytes for these studies. RESULTS: The hepatocyte-like cells have been developed from hTERT plus Bmi-1-immortalized human mesenchymal stem cells to substitute the primary hepatocytes. The hepatocyte-like cells had polygonal morphology and steadily produced albumin, glycogen, urea and UGT1A1 beyond 6 months while maintaining proliferative capacity. Although these hepatocyte-like cells had low basal expression of CYP450 isotypes, their expressions could be extensively up regulated to 80 folds upon the exposure to enzyme inducers. Their inducibility outperformed the classical HepG2 cells. CONCLUSION: The hepatocyte-like cells contained the markers of hepatocytes including CYP450 isotypes. The high inducibility of CYP450 transcripts could serve as a sensitive model for profiling xenobiotic-induced expression of CYP450.

The Protective Effect of Polyherbal Formulation, Harak Formula, on UVA-Induced Photoaging of Human Dermal Fibroblasts and Mouse Skin via Promoting Nrf2-Regulated Antioxidant Defense.

Polyherbal formulation combining multiple herbs is suggested to achieve enhanced therapeutic effects and reduce toxicity. Harak herbal formula (HRF) extracts were proposed to regulate skin responses to UVR through their ability to suppress UVA-induced matrix metalloproteinase-1 (MMP-1) and pigmentation via promoting antioxidant defenses in in vitro models. Therefore, natural products targeting Nrf2 (nuclear factor erythroid 2-related factor 2)-regulated antioxidant response might represent promising anti-photoaging candidates. Hesperetin (HSP) was suggested as a putative bioactive compound of the HRF, as previously shown by its chemical profiling using the liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). In this study, we explored the anti-photoaging effects of HRF extracts and HSP on normal human dermal fibroblasts (NHDFs) and mouse skin exposed to UVA irradiation. Pretreatment of NHDFs with HRF extracts and HSP protected against UVA (8 J/cm2)-mediated cytotoxicity and reactive oxygen species (ROS) formation. The HRF and HSP pretreatment also attenuated the UVA-induced MMP-1 activity and collagen depletion concomitant with an upregulation of Nrf2 activity and its downstream genes (GST and NQO-1). Moreover, our findings provided the in vivo relevance to the in vitro anti-photoaging effects of HRF as topical application of the extracts (10, 30 and 100 mg/cm2) and HSP (0.3, 1, and 3 mg/cm2) 1 h before UVA exposure 3 times per week for 2 weeks (a total dose of 60 J/cm2) mitigated MMP-1 upregulation, collagen loss in correlation with enhanced Nrf2 nuclear accumulation and its target protein GST and NQO-1 as well as reduced 8-hydroxy-2'-deoxyguanosine (8-OHdG) in irradiated mouse skin. Thus, our findings revealed that HRF extracts and HSP attenuated UVA-induced photoaging via upregulating Nrf2, together with their abilities to reduce ROS formation and oxidative damage. Our study concluded that the HRF and its bioactive ingredient HSP may represent potential candidates for preventing UVA-induced photoaging via restoration of redox balance.