Alpha-Glucosidase Inhibition, Antioxidant Activities, and Molecular Docking Study of Krom Luang Chumphon Khet Udomsak, a Thai Traditional Remedy.

Krom Luang Chumphon Khet Udomsak remedy (KKR) has traditionally been used as an alternative treatment, particularly for hyperglycemia; however, its therapeutic efficacy has not been scientifically validated. Thus, this study aims to investigate the potential inhibitory and antioxidant effects of α-glucosidase enzyme and characterize the chemical profile of KKR extracts using gas chromatography-mass spectrometry (GC-MS). The investigation highlights both KKR extracts as potent inhibitors of α-glucosidase, with the ethanolic extract of KKR (KKRE) displaying an IC50 value of 46.80 µg/mL and a noncompetitive mode of action. The combination of ethanolic and aqueous extracts of KKR (KKRE and KKRA, respectively) with acarbose exhibited a synergistic effect against the α-glucosidase. The KKRE extract displayed strong scavenging effects in the DPPH assay (IC50 156.3 µg/mL) and contained significant total phenolic (172.82 mg GAE/g extract) and flavonoid (77.41 mg QE/g extract) contents. The major component of KKRE is palmitic acid (15.67%). Molecular docking revealed that the major compounds interacted with key amino acid residues (ASP215, GLU277, HIS351, ASP352, and ARG442), which are crucial for inhibiting α-glucosidase. Notably, campesterin had a more significant influence on α-glucosidase than acarbose, with low binding energy. These findings underscore the significance of KKR in traditional medicine and suggest that it is promising treatment for diabetes mellitus. Further studies using animal model will provide valuable insights for advancing this research.

Experimental design approach for the quantitative analysis of multicomponents by single marker and HPLC fingerprinting of Thunbergia laurifolia aqueous extract.

INTRODUCTION: Thunbergia laurifolia is used in traditional Thai medicine to reduce fever and treat mouth ulcers. However, the quantitative analysis of chemical markers has not yet been officially defined. OBJECTIVE: The objective of this study is to develop a high-performance liquid chromatography (HPLC) method using a design of experiment (DoE) for the quantitative analysis of multicomponents by single marker (QAMS) and fingerprinting of the T. laurifolia aqueous extract. MATERIALS AND METHODS: Critical variables were screened using a two-level fractional factorial design, followed by the optimization of the selected variables using a central composite design. The validated method was applied for quality assessment based on QAMS and fingerprinting of the extract. RESULTS: Optimum conditions of DoE for the analysis of caffeic acid, vicenin-2, and rosmarinic acid were determined. The relative correction factors for caffeic acid and vicenin-2 were calculated using rosmarinic acid as an internal reference standard, and their contents in 30 samples were determined. The differences between the external standard method (ESM) and QAMS were compared. No significant difference was observed in the quantitative determination, proving the consistency QAMS and ESM. HPLC fingerprints of T. laurifolia were established with 8 of 12 characteristic peaks that were structurally characterized using HPLC-diode array detection-electrospray ionization/tandem mass spectrometry. The similarity of the fingerprints in all samples was ≥0.74, and the pattern recognition of the characteristic peaks was satisfied. CONCLUSION: The proposed method efficiently detected multiple components of the T. laurifolia extract. Thus, the method is beneficial in providing references for enhancing the quality control of other herbal medicines.

A Potential Therapeutic Approach According to Traditional Thai Medicine Theory for Post-COVID-19 Syndrome Treatment: An Observational Retrospective Clinical Study.

Background: Most post-COVID-19 patients had physiological health problems, which differed for each patient. The potential treatment for post-COVID-19 syndrome must require multi-disciplinary approaches that focus on individualized treatment. Therefore, traditional Thai Medicine (TTM) clinical practice guidelines for post-COVID-19 treatment are developed by the Traditional Thai Medicine Hospital, Prince of Songkla University, to care for post-COVID-19 patients. Objective: This current study aimed to investigate the effect of the guidelines on post-COVID-19 patient's physical health. Methods: This research was a retrospective clinical study, carried out from June 2022 to July 2022. Setting: The study was conducted at the Traditional Thai Medicine Hospital, Prince of Songkla University, and La Flora Khao Lak TTM clinic, Thailand. Participants: Data (n=17) were collected from the medical record. They were included if they met the following criteria: patients were more than 18 years old, had a medical history of COVID-19 disease, had fully recovered from the disease (ATK for COVID-19 is negative) at least a week before starting the guideline, and had complete medical information such as post-COVID-19 symptoms, vital signs, pain score, cough severity score, and quality of life examination. Intervention: Approaches for post-COVID-19 treatment included herbal Thai medicine recipes, Thai massage, herbal steam inhalation, herbal hot steam, herbal hot water immersion, acupressure massage for breathing stimulation, and a breathing exercise. All procedures were applied for 7 days continually or until they have recovered. Primary Outcome Measures: The effects of TTM approaches on the symptoms of post-COVID-19 patients on pain score, quality of life, cough severity were measured using a visual analog score (VAS), SF-36 questionnaire, and cough severity index (CSI), respectively. Results: The average duration time of post-COVID-19 syndrome was 25.76 days, and the most common symptom was muscle pain/stress (10; 71.43%). Yahom-Navakot (6; 42.86%) is the most common recipe used for the treatment. Interestingly, the pain score was significantly reduced by 3 days of the treatment (4.88±2.03 vs 2.29±2.08 ). Moreover, the cough severity index (12.86±11.55 vs 3.31±6.38) was significantly lessened by 7 days of the treatment. Remarkably, most symptoms vanished entirely after 7 days of the intervention, leading to a better quality of life for the patients (53.24±22.15 vs 65.59±23.64). Conclusions: The TTM clinical practice guideline for treating post-COVID-19 syndrome effectively improved the overall physical health capacity and symptoms associated with post-COVID-19 syndrome. Therefore, the implementation of this guideline, together with other approaches, could strengthen the treatment for post-COVID-19 to be more effective.

Anti-inflammatory effects of the Thai herbal remedy Yataprasen and biflavonoids isolated from Putranjiva roxburghii in RAW264.7 macrophages.

ETHNOPHARMACOLOGICAL RELEVANCE: Yataprasen is a topical Thai herbal remedy for the treatment of musculoskeletal pain and is included in Kumpe Thart Phra Narai, the first Thai textbook of traditional medicine. The herbal preparation is made from a hydroethanolic extract of a mixture of 13 medicinal plants, of which Putranjiva roxburghii Wall. leaves are the major ingredient. AIM OF THE STUDY: In this study, we investigated the underlying mechanism of action for the anti-inflammatory effects of the Yataprasen remedy, its main ingredients, and the phytochemicals isolated from P. roxburghii leaves. MATERIALS AND METHODS: The anti-inflammatory effects of the Yataprasen remedy, along with its main ingredients, including the leaves of Baliospermum solanifolium (Burm.) Suresh, Melia azedarach L., P. roxburghii, Senna siamea (Lam.) Irwin & Barneby, and Tamarindus indica L. were determined by measuring prostaglandin E2 (PGE2) secretion, nitric oxide (NO) production, and the synthesis of inflammatory biomarkers in lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells. The active ingredients of the P. roxburghii leaves were separated by chromatography and spectroscopic measurements were used to identify their chemical structures. RESULTS: Ethanol extracts of the Yataprasen remedy and some of its ingredients significantly suppressed LPS-induced PGE2 secretion and NO production in a dose-dependent manner. Treatment of RAW264.7 cells with ethanolic extracts of the Yataprasen remedy (50 µg/mL) significantly inhibited LPS-induced mRNA expression of TNF-α, COX-2, iNOS, and NF-κB. Among the plant ingredient extracts, P. roxburghii leaf extract exhibited the highest inhibitory effects on LPS-induced TNF-α and iNOS expression. Moreover, T. indica leaf extract showed the highest activity on the inhibition of LPS-induced COX-2 and NF-κB expression. Putraflavone, podocarpusflavone A, and amentoflavone were isolated biflavonoids from P. roxburghii leaf extract and showed the inhibitory effects on LPS-induced PGE2 secretion and NO synthesis in RAW264.7 cells. Of the isolated biflavonoids, amentoflavone exhibited the strongest anti-inflammatory activity by inhibiting the expression of TNF-α, COX-2, and iNOS. CONCLUSION: The results support reported the anti-inflammatory effects of the Yataprasen remedy, which are associated with the downregulation of proinflammatory mediators. P. roxburghii, along with its biflavonoids, are the impact components that contribute to the anti-inflammatory effects of the herbal remedy.

Thai traditional medicines reduce CD147 levels in lung cells: Potential therapeutic candidates for cancers, inflammations, and COVID-19.

ETHNOPHARMACOLOGICAL RELEVANCE: The cluster of differentiation 147 (CD147) is identified as the signaling protein relevant importantly in various cancers, inflammations, and coronavirus disease 2019 (COVID-19) via interacting with extracellular cyclophilin A (CypA). The reduction of CD147 levels inhibits the progression of CD147-associated diseases. Thai traditional medicines (TTMs): Keaw-hom (KH), Um-ma-ruek-ka-wa-tee (UM), Chan-ta-lee-la (CT), and Ha-rak (HR) have been used as anti-pyretic and anti-respiratory syndromes caused from various conditions including cancers, inflammations, and infections. Thus, these medicines would play a crucial role in the reduction of CD147 levels. AIM OF THE STUDY: This article aimed to investigate the effects of KH, UM, CT, and HR for reducing the CD147 levels through in vitro study. Additionally, in silico study was employed to screen the active compounds reflexing the reduction of CD147 levels. MATERIALS AND METHODS: The immunofluorescent technique was used to evaluate the reduction of CD147 levels in human lung epithelial cells (BEAS-2B) stimulated with CypA for eight extracts of KH, UM, CT, and HR obtained from water decoction (D) and 70% ethanol maceration (M) including, KHD, UMD, CTD, HRD, KHM, UMM, CTM, and HRM. RESULTS: UM extracts showed the most efficiency for reduction of CD147 levels in the cytoplasm and perinuclear of BEAS-2B cells stimulated with CypA. Phenolic compounds composing polyphenols, polyphenol sugars, and flavonoids were identified as the major chemical components of UMD and UMM. Further, molecular docking calculations identified polyphenol sugars as CypA inhibitors. CONCLUSIONS: UMD and UMM are potential for reduction of CD147 levels which provide a useful information for further development of UM as potential therapeutic candidates for CD147-associated diseases such as cancers, inflammations, and COVID-19.

Anti-proliferative Effects of Pinocembrin Isolated From Anomianthus dulcis on Hepatocellular Carcinoma Cells.

BACKGROUND: Hepatocellular carcinoma (HCC) is the most prevalent primary liver cancer. Anomianthus dulcis (Dunal) J.Sinclair (syn. Uvaria dulcis) has been used in Thai traditional medicine in various therapeutic indications. Phytochemical constituents of A. dulcis have been isolated and identified. However, their effects on liver cancer and the associated mechanisms have not been elucidated. METHODS: Dry flowers of A. dulcis were extracted using organic solvents, and chromatographic methods were used to purify the secondary metabolites. The chemical structures of the pure compounds were elucidated by analysis of spectroscopic data. Cytotoxicity against HCC cells was examined using SRB assay, and the effects on cell proliferation were determined using flow cytometry. The mechanisms underlying HCC inhibition were examined by molecular docking and verified by Western blot analysis. RESULTS: Among 3 purified flavonoids, pinocembrin, pinostrobin, and chrysin, and 1 indole alkaloid (3-farnesylindole), only pinocembrin showed inhibitory effects on the proliferation of 2 HCC cell lines, HepG2 and Li-7, whereas chrysin showed specific toxicity to HepG2. Pinocembrin was then selected for further study. Flow cytometric analyses revealed that pinocembrin arrested the HCC cell cycle at the G1 phase with a minimal effect on cell death induction. Pinocembrin exerted the suppression of STAT3, as shown by the molecular docking on STAT3 with a better binding affinity than stattic, a known STAT3 inhibitor. Pinocembrin also suppressed STAT3 phosphorylation at both Tyr705 and Ser727. Cell cycle regulatory proteins under the modulation of STAT3, namely cyclin D1, cyclin E, CDK4, and CDK6, are substantially suppressed in their expression levels. CONCLUSION: Pinocembrin extracted from A. dulcis exerted a significant growth inhibition on HCC cells via suppressing STAT3 signaling pathways and its downstream-regulated genes.

The interaction of Suk-Saiyasna remedy with GABAA and CB1 receptor-targeting drugs: Enhancing hypnotic and sedative effects in in vivo models.

The Suk-Saiyasna remedy, an herbal treatment, was historically used but ceased due to its cannabis content. After a relaxation of drug control laws in Thailand, its use re-emerged. This study examines the Suk-Saiyasna remedy's impact on rodent behavior and its receptor effects. This study was conducted to assess the sedative-like effects of the remedy on mice. The mice were divided into groups receiving 0.6, 3, 30, and 60 mg/kg extracts, with negative controls for comparison. We also investigated the impact on receptors, utilizing negative controls and pretreatment with receptor blockers, followed by either a negative control or a 60 mg/kg extract. Furthermore, this study investigated the behavioral aspects of mice, including anxiolytic effects, antidepressant-like effects, and motor coordination, utilizing the elevated plus-maze, open-field, and rotarod performance tests. The Suk-Saiyasna remedy (P < 0.05) decreased significantly in the latent period and increased sleeping time in the treated groups. Moreover, the Suk-Saiyasna remedy also showed efficacy in reaction to GABAA receptors and cannabinoid CB1 receptors (P < 0.05). In addition, positive effects were observed regarding the animal behavior in the arena, as the animal activity, behavior, and muscle coordination were reduced (P < 0.05). The Suk-Saiyasna remedy may be involved in a sedative-hypnotic-like effect in rodents under normal conditions through the modulation of GABAergic neurons and induction of the cannabinoid CB1 receptor mechanism.

Anti-inflammatory potential of a Thai traditional remedy called Prabchompoothaweep in an animal model of acute and sub-acute inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: Prabchompoothaweep (PCT) is a Thai remedy which is composed of 23 herbs and has been added onto the National List of Essential Medicines (NLEM) of Thailand. This remedy has been used to treat allergic rhinitis and asthma in Thai traditional medicine for many years. Furthermore, a few studies have reported anti-allergic, anti-malarial, anti-inflammatory and antioxidant activities. AIM OF THE STUDY: This study aims to evaluate anti-inflammatory activity of PCT extract in an animal model. MATERIALS AND METHODS: The animal model of acute inflammation was studied over a 24-h period, utilizing the method of carrageenan-induced paw oedema. In addition, sub-acute inflammation was examined over 7 days, using the formalin-induced paw oedema method. The treatment groups received PCT extracts, via the oral route, at 1-h prior to injection and then the sub plantar of the rat right paw was injected with the named substances to generate paw oedema. The paw thickness was measured by vernier caliper at regular intervals after injection. At the end of experiment, the blood and paw tissues were collected for measurement of pro-inflammatory cytokines and histological examination respectively. RESULTS: In acute inflammation, all doses of PCT extract (250, 500 and 1000 mg/kg p.o.) significantly reduced paw thickness after the first 3 h in a dose-dependent manner and the percentage of inhibition was 38.7%, 47.8% and 49.5% respectively. The pro-inflammatory cytokines, including TNF-α and IL-1ß, statistically decreased with all doses of the extracts. However, the histological examination did not reveal significant results due to the short time duration. As regards to sub-acute inflammation, all doses of PCT extract significantly reduced paw thickness with 12.78%, 23.64% and 35.78%, in a dose dependent manner. Also, the pro-inflammatory cytokines (TNF-α and IL-1ß) significantly decreased at day 7. Interestingly, the histological examination of paw tissue demonstrated reductions of mononuclear infiltrations of inflammatory cells, this was observed in the group receiving PCT extracts, also in a dose-dependent manner. CONCLUSION: Therefore, PCT exerted anti-inflammatory activity in an animal model of acute and sub-acute inflammation, suggesting that it could be used as a new source for treatment of inflammatory diseases.

Phytochemical Profiling and Antioxidant Capacity of Traditional Plants, Northern Thailand.

Traditional plants have played a significant role in human culture and medicine throughout history. These plants have the capability to synthesize a diverse range of chemical compounds that serve essential biological functions. This study's objective was to analyze the phytochemical composition of five traditional plants, namely Emilia sonchifolia, Chloranthus erectus, Caesalpinia mimosoides, Acacia concinna, and Tacca chantrieri, native to northern Thailand, using LC-QTOF/MS analysis and assess their potential bioactivity through various assays including DPPH radical scavenging activity, ABTS radical scavenging activity, ferric ion reducing antioxidant power, total phenolic compounds, and total flavonoid content. The findings revealed the presence of natural bioactive compounds in each plant extract, which exhibited pharmacological activity. Notably, Caesalpinia mimosoides displayed the highest antioxidant capacity across all plant extracts (IC50 in DPPH with the methanol extract was 0.03 and 898.18 mg AAE/100 g with the ethanol extract), along with elevated levels of total phenolic and flavonoid content, which showed the highest TFC at 46.79 µgRE/g in the methanol extract. In conclusion, traditional plants possess notable biological constituents and antioxidant properties, suggesting their potential for bioactive applications. Based on these findings, these indigenous plants can serve as a valuable resource in traditional medicine, offering the possibility of uncovering new products with similar capabilities and additional therapeutic attributes worthy of future exploration.

Clinacanthus nutans genetic diversity and its association with anti-apoptotic, antioxidant, and anti-bacterial activities.

Clinacanthus nutans (Burm. f.) Lindau has been extensively utilized in Thai folk medicine. However, there has been no prior exploration of its genetic diversity or its correlation with biological activity and phytochemical profiles. Herein, a total of 10 samples of C. nutans were collected from different geographic locations in different environments of Thailand, encompassing Northern, Northeastern, and Central regions. The genetic diversity study using sequence-related amplified polymorphism (SRAP) markers showed that all C. nutans samples were closely related, as indicated by UPGMA cluster analysis. When comparing the biological activities of C. nutans extracts, our findings demonstrated that those sourced from Northern Thailand exhibited the most potent activity in reducing lipopolysaccharide-inducing cell death, as accessed by cell viability assay. Furthermore, they showed remarkable antioxidant and antibacterial activities against Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. High-performance liquid chromatography (HPLC) analysis of phytochemical profiles revealed consistent chromatography peak patterns across all C. nutans extracts. However, they exhibited varying levels of phenolic contents, as judged by the Folin-Ciocalteu assay, which positively correlated with their observed activities. In conclusion, this study highlights the limited genetic variation within C. nutans population in Thailand. Furthermore, it underscores the association between the biological activity and the total phenolic contents which might be mainly impacted by environmental conditions.

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.

Predictors of and healthcare utilisation of depressive symptoms among middle-aged and older adults in Thailand: a national cross-sectional community-based study in 2015.

OBJECTIVE: The prevalence of depressive symptoms and healthcare utilisation among the ageing population in Thailand is unclear. The study assessed the predictors of and healthcare utilisation of depressive symptoms among an ageing population in Thailand. DESIGN: Cross-sectional, population-based study. SETTING: National sample of people aged 45 years and older in Thailand from the 2015 Health, Ageing and Retirement in Thailand (HART) study. PARTICIPANTS: The sample included 5135 individuals (≥45 years) with complete depressive symptoms data from the 2015 HART study. OUTCOME MEASURES: Depressive symptoms were measured with the Centre for Epidemiological Studies Depression Scale. Logistic regression was used to estimate the associations between sociodemographic factors, health factors and depressive symptoms, and the associations between depressive symptoms and different types of healthcare utilisation. RESULTS: The study population included 5135 participants with a median age of 66 years. The prevalence of depressive symptoms was 13.9%. The proportion of past 2 year hospital admission was 12.8%, hospital outpatient 49.7%, health centre outpatient 26.2%, private clinic 8.4%, traditional medical practitioner 2.8%, medical home visit 4.9% and past-year medical check-up 50.3%. In adjusted logistic regression analysis, low income, residing in the Southern region, history of a cardiovascular disease, brain diseases and/or psychiatric problems and functional disability were positively associated with depressive symptoms. Male sex, being a Buddhist, urban residence, high religious involvement, moderate and high physical activity were negatively associated with depressive symptoms. In fully adjusted logistic regression models, depressive symptoms were positively associated with hospital inpatient utilisation, hospital outpatient utilisation, health centre utilisation and utilisation of a traditional medicine practitioner. CONCLUSIONS: More than one in 10 participants had depressive symptoms. Chronic conditions, sociodemographic factors, physical inactivity and low religious involvement increased the odds of depressive symptoms. Furthermore, depressive symptoms increased the odds of conventional and traditional healthcare utilisation.

In vitro callus induction and evaluation of antioxidant activity of Rhinacanthus nasutus (L.) Kurz.

Rhinacanthus nasutus (L.) Kurz is used in Thai traditional medicine for the treatment of skin diseases, ringworm, and eczema. This research studied the effects of cytokinin and auxins on callus induction and evaluated antioxidant activity of R. nasutus. Nodes, young, and mature leaf explants were cultured on MS medium containing 0, 1, 2, 3, and 4 mg/l kinetin (6-furfurylaminopurine) and 0, 1 mg/l 1-naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), and 2,4-dichlorophenoxyacetic acid (2,4-D) for 6 weeks to induce callus. Calli derived from nodes, young and mature leaves, and other plant parts were ultrasonically extracted with methanol to determine total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity by ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhtdrazyl (DPPH), and 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) assays. Nodal explants on MS medium containing 1 mg/l kinetin combined with 1 mg/l 2,4-D were most efficient in callus production with the average fresh and dry weight per explant of 2.29 ± 0.14 and 0.18 ± 0.01 g, respectively. Addition of kinetin combined with NAA or 2,4-D had a positive effect on callus induction from young and mature leaf explants. The leaf extract showed the highest TPC, TFC, FRAP, and IC50 of DPPH and ABTS assays (ca 113 mg GAE/g extract, 45 mg QE/g extract, 121 mg TE/g extract, 53 µg/ml and 14 µg/ml, respectively), followed by callus derived from nodes. Overall, phenolic content was higher than flavonoid content. A strong positive correlation was found between FRAP assay, TPC (r = 0.973), and TFC (r = 0.798), indicating that phenolic and flavonoid compounds are responsible for antioxidant activity of R. nasutus.

Phytochemical Screening on Phenolic, Flavonoid Contents, and Antioxidant Activities of Six Indigenous Plants Used in Traditional Thai Medicine.

The antioxidant activity of a traditional Thai formula has been studied and compared to each plant. The formula comprised the roots of Caesalpinia digyna Rottler, Huberantha cerasoides (Roxb.) Benth), Oxyceros horridus Lour, Antidesma ghaesembilla Gaerth, Combretum quadrangulare Kurz, and Ziziphus cambodiana Pierre. The stem was also studied in comparison. The ethanolic extract from each plant part and the mixed plants mimicking the traditional formula were prepared and investigated for antioxidant capability in vitro via DPPH radical scavenging and ferric-reducing antioxidant power assays. The phytochemical constituents were determined by chemical screening, total phenolic (TPC) and flavonoid contents (TFC), and high-performance liquid chromatography. The relationship between antioxidant activity and the contributed phytochemicals was determined using correlation analysis and principal component analysis (PCA). Results showed that extracts from both parts of the plant formula showed the highest antioxidant activity compared to a single plant extract. Among the six plants, C. digyna exhibited the highest TPC and antioxidant activity. TPC had a strong positive correlation with antioxidant activity. PCA revealed that gallic acid contributed to the antioxidant activity. In conclusion, the ethanolic extracts of the traditional formula and C. digyna have the potential for further chemical characterization and study related to antioxidant activity.

Thai traditional massage modulates urinary MCP-1 and relevant inflammatory biomarkers in lower urinary tract symptom patients.

Lower urinary tract symptoms (LUTS) resulting from benign prostatic hyperplasia are a common complaint among elderly men worldwide. Our previous study reported alleviative efficacy of Thai traditional massage (TTM) on LUTS patients. However, underlying mechanism at cellular level remained elusive. Herein, we investigated the effect of TTM on urinary monocyte chemotactic protein-1 (MCP-1) and associative inflammatory biomarkers. Forty-three patients were randomized into two groups: Tamsulosin (n = 23) and TTM (n = 20). The urinary MCP-1 and interferon-gamma (IFN-γ) levels as well as gene expression levels of MCP-1, Chemotactic protein receptor 2b (CCR2b), IFN-γ, interleukin-1 beta (IL-1ß), and transforming growth factor-beta 1 (TGF-ß1) were evaluated before and after a four-week treatment. The urinary MCP-1 and IFN-γ levels as well as gene expression levels of MCP-1, CCR2b, IFN-γ, IL-1ß, and TGF-ß1 were evaluated before and after treatment with Tamsulosin or TTM group. Urinary MCP-1 and IFN-γ levels and the expression levels of five genes from sedimented urine samples were measured using Enzyme-Linked Immunosorbent Assay and quantitative Reverse Transcription Polymerase Chain Reaction, respectively. We observed significant (p < 0.05) reduction in the ratio of urinary MCP-1 and creatinine (Cr); MCP-1/Cr levels in subjects given only TTM. There were no significant differences (p < 0.05) in IFN-γ/Cr levels in both groups. TTM group down-regulated the expression of IFN-γ whereas up-regulated IL-1ß and TGF-ß1 mRNA. Our findings suggested TTM had alleviative effects in LUTS patients, which were partially mediated by a reduction of urinary inflammatory cytokines and inflammatory gene expression.

Study of Optimal Conditions to Grow Thai Ganoderma, Fruiting Test, Proximate and Their Alpha Glucosidase Inhibitory Activity.

Ganoderma (Ganodermataceae) has a worldwide distribution and has been widely used in traditional medicines. In this study, we report wild strains of Ganoderma that include two G. sichuanense and one G. orbiforme from northern Thailand. Optimal conditions for mycelium growth were ensured. The most favourable medium was potato sucrose agar for G. sichuanense and oatmeal agar for G. orbiforme and at 25 °C and 30 °C and pH 4-8. All types of cereal grains can be used to promote the growth of the mycelia of Ganoderma species. Fruiting tests were performed. All strains of Ganoderma produce fruiting bodies successfully in bag culture at 28 ± 1 °C with 75-85% relative humidity. Only G. orbiforme produced fruiting bodies in field cultivation at the laboratory scale. In the first flush yields, the G. sichuanense strain MFLUCC 22-0064 gave better production (the B.E was 152.35 ± 6.98 g). This study is the first to document the bag and field cultivation of wild Thai G. orbiforme. Ganoderma species are revealed to contain high amounts of fiber (47.90-52.45% d.b.), protein (12.80-14.67% d.b.), fat (4.90-5.70% d.b.), and carbohydrates (3.16-4.02% d.b.). Additionally, G. sichuanense and G. orbiforme were preliminarily screened for biological activity for inhibition of alpha-glucosidase enzyme activity. The IC50 values of G. orbiforme (MFLUCC 22-0066) was 105.97 ± 1.36 µg/mL and G. sichuanense (MFLUCC 22-0064) was 126.94 ± 0.87 µg/mL. Both strains had better inhibition than acarbose (168.18 ± 0.89 µM). These results on wild strains of Ganoderma will be useful for further studies on the applications of Ganoderma. Later the species can be introduced to domestic markets for cultivation and medicinal use.

In vitro evaluation of the anti­breast cancer properties and gene expression profiles of Thai traditional formulary medicine extracts.

Breast cancer is a leading cause of cancer-related deaths worldwide. Moreover, standard treatments are limited, so new alternative treatments are required. Thai traditional formulary medicine (TTFM) utilizes certain herbs to treat different diseases due to their dominant properties including anti-fungal, anti-bacterial, antigenotoxic, anti-inflammatory and anti-cancer actions. However, very little is known about the anti-cancer properties of TTFM against breast cancer cells and the underlying molecular mechanism has not been elucidated. Therefore, the present study, evaluated the metabolite profiles of TTFM extracts, the anti-cancer activities of TTFM extracts, their effects on the apoptosis pathway and associated gene expression profiles. Liquid chromatography with tandem mass spectroscopy analysis identified a total of 226 compounds within the TTFM extracts. Several of these compounds have been previously shown to have an anti-cancer effect in certain cancer types. The MTT results demonstrated that the TTFM extracts significantly reduced the cell viability of the breast cancer 4T1 and MDA-MB-231 cell lines. Moreover, an apoptosis assay, demonstrated that the TTFM extracts significantly increased the proportion of apoptotic cells. Furthermore, the RNA-sequencing results demonstrated that 25 known genes were affected by TTFM treatment in 4T1 cells. TTFM treatment significantly up-regulated Slc5a8 and Arhgap9 expression compared with untreated cells. Moreover, Cybb, and Bach2os were significantly downregulated after TTFM treatment compared with untreated cells. Reverse transcription-quantitative PCR demonstrated that TTFM extract treatment significantly increased Slc5a8 and Arhgap9 mRNA expression levels and significantly decreased Cybb mRNA expression levels. Moreover, the mRNA expression levels of Bax and Casp9 were significantly increased after TTFM treatment in 4T1 cells compared with EpH4-Ev cells. These findings indicated anti-breast cancer activity via induction of the apoptotic process. However, further experiments are required to elucidate how TTFM specifically regulates genes and proteins. This study supports the potential usage of TTFM extracts for the development of anti-cancer drugs.

Curcumin and proton pump inhibitors for functional dyspepsia: a randomised, double blind controlled trial.

OBJECTIVE: To compare the efficacy of curcumin versus omeprazole in improving patient reported outcomes in people with dyspepsia. DESIGN: Randomised, double blind controlled trial, with central randomisation. SETTING: Thai traditional medicine hospital, district hospital, and university hospitals in Thailand. PARTICIPANTS: Participants with a diagnosis of functional dyspepsia. INTERVENTIONS: The interventions were curcumin alone (C), omeprazole alone (O), or curcumin plus omeprazole (C+O). Patients in the combination group received two capsules of 250 mg curcumin, four times daily, and one capsule of 20 mg omeprazole once daily for 28 days. MAIN OUTCOME MEASURES: Functional dyspepsia symptoms on days 28 and 56 were assessed using the Severity of Dyspepsia Assessment (SODA) score. Secondary outcomes were the occurrence of adverse events and serious adverse events. RESULTS: 206 patients were enrolled in the study and randomly assigned to one of the three groups; 151 patients completed the study. Demographic data (age 49.7±11.9 years; women 73.4%), clinical characteristics and baseline dyspepsia scores were comparable between the three groups. Significant improvements were observed in SODA scores on day 28 in the pain (-4.83, -5.46 and -6.22), non-pain (-2.22, -2.32 and -2.31) and satisfaction (0.39, 0.79 and 0.60) categories for the C+O, C, and O groups, respectively. These improvements were enhanced on day 56 in the pain (-7.19, -8.07 and -8.85), non-pain (-4.09, -4.12 and -3.71) and satisfaction (0.78, 1.07, and 0.81) categories in the C+O, C, and O groups, respectively. No significant differences were observed among the three groups and no serious adverse events occurred. CONCLUSION: Curcumin and omeprazole had comparable efficacy for functional dyspepsia with no obvious synergistic effect. TRIAL REGISTRATION NUMBER: TCTR20221208003.

Gastroprotective effects of Pikad Tri-phol-sa-mut-than herbal remedy on multiple gastric ulcer models in rats.

In Thai traditional medicine, Pikad Tri-phol-sa-mut-than has long been used to alleviate gastrointestinal symptoms, renal disease, inflammation-related disorders, and severe malady. This recipe is composed of dried fruits of Morinda citrifolia L., Coriandrum sativum L., and Aegle marmelos (L.) Corrêa. The aim of this study was to assess the anti-gastric ulcer property of the water extract of Pikad Tri-phol-sa-mut-than (TS), using various animal models with different inducers, including restraint water immersion stress, indomethacin, and ethanol/hydrochloric acid (EtOH/HCl). Its mechanisms of anti-gastric ulcer actions were also elucidated using both in vitro and in vivo experiments. When compared with the control groups, the oral pretreatment of TS at the doses of 150, 300 and 600 mg/kg significantly reduced the gastric ulcer formation in all models. It was also found that TS at the dose of 600 mg/kg could increase gastric wall mucus in rats but could not produce the significant reduction of the gastric volume or total acidity of gastric content. Results from hematoxylin and eosin (H&E) and Periodic acid-Schiff (PAS) staining examinations of gastric tissues confirmed that TS visibly reduced gastric mucosal damage, while immunohistochemistry revealed that TS remarkably suppressed the protein expression of Bcl-2-associated X (BAX), a regulator of apoptosis, compared to those of the control group. The DPPH, ABTS, and FRAP assays showed antioxidant effects of TS. All of these findings demonstrated that TS has gastroprotective effects, which may be related to the increase in the gastric wall mucus secretion, not anti-secretory activity, as well as its antioxidant and antiapoptotic activities.

Formulation design and physicochemical evaluation of an anti-inflammatory hydrogel patch containing Crinum asiaticum L. extract.

Background and purpose: Crinum asiaticum L. has long been used in Thai traditional medicine to treat osteoarthritis and inflammation by placing it on painful areas without further formulation design which is suboptimal for therapeutic use. Thus, this research aims to formulate a topical hydrogel patch containing C. asiaticum L. extracts (CAE) for anti-inflammatory effects. Experimental approach: The hydrogel patches are made from carrageenan, locust bean gum, with glycerin as a plasticizer and contain CAE formulated by using response surface methodology based on Box-Behnken design for design, determination of the effect of independent factors on the tensile strength, and optimization of the hydrogel patch formulation. In vitro release and skin permeation studies using a modified Franz diffusion cell and anti-inflammatory activity were evaluated. Findings/Results: The optimized CAE hydrogel patch showed a good correlation between predicted and observed tensile strength values and exerted its maximum cumulative lycorine release and permeation at 69.38 ± 2.78% and 48.51 ± 0.45%, respectively which were fit to Higuchi's kinetic model. The release rates were found to decrease with an increase in the polymer proportion of carrageenan and locust bean gum. In addition, the patch exerted potent in vitro anti-inflammatory activity with an IC50 value of 21.36 ± 0.78 µg/mL. Conclusion and implication: The optimized CAE hydrogel patch application was successfully formulated with excellent mechanical properties, cumulative release, permeation, and anti-inflammatory effects. Thus, it has the potential to be further developed as a herbal application to relieve pain and inflammation. The in vivo anti-inflammatory effect of this delivery system should be further investigated.