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.

Antibacterial, anti-inflammatory and antioxidant activities of Mahanintangtong and its constituent herbs, a formula used in Thai traditional medicine for treating pharyngitis.

BACKGROUND: Mahanintangtong is listed in the Thailand's National List of Essential Medicines (NLEM). It is used to treat non-specific fevers and illnesses such as pharyngitis and chickenpox. In this study, we investigated the biological activities of the different medicinal plants used in the Mahanintangtong formula. METHODS: The plant materials were extracted by maceration and decoction. Antimicrobial activity, assessed by disc diffusion method, the minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) were compared with commercially available standard antibiotics. To elucidate the anti-inflammatory mechanisms, inhibition of nitric oxide (NO), tumor necrosis factor α (TNF-α), and interleukin 6 (IL-6) production was tested by Griess and ELISA techniques. Antioxidant activity was measured by ABTS and DPPH scavenging assays. RESULTS: The extracts with the best antimicrobial activities were carbonized Tectona grandis showing against Streptococcus pyogenes, Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. The ethanol extract of Dracaena loureiroi wood exhibited the highest NO and IL-6 inhibitory activity with IC50 values of 9.42 ± 1.81 and 12.02 ± 0.30 µg/mL, respectively. The ethanol extract of Pogostemon cablin had the highest TNF-α inhibitory with IC50 values of 10.68 ± 0.02 µg/mL. In anti-free radical testing, the ethanol extract of D. loureiroi displayed high antioxidant activity by both ABTS and DPPH assays. CONCLUSION: The ethanol extracts from carbonized T. grandis and Mahanintangtong showed good antimicrobial activity, especially against S. pyogenes, and good anti-inflammatory activity. These findings are relevant to the pathogenesis of pharyngitis and justify additional studies to see if Mahanintangtong could have clinical utility.

Antioxidant activities and phytochemical constituents of Antidesma thwaitesianum Müll. Arg. leaf extracts.

OBJECTIVE: To investigate the antioxidant activities as well as phytochemical constituents of Antidesma thwaitesianum Müll. Arg. leaf extracts. METHODS: The leaves of A. thwaitesianum were extracted using three different methods: blending with distilled water, maceration with ethanol and decoction. The chemical antioxidant activity of the plant leaf extracts was evaluated using 2,2-diphenyl-1-picryhydrazyl (DPPH) radical and 2,2'-azinobis(3-ethylbenzo-thiazoline-6-sulfonic acid) diammonium salt (ABTS⁺) radical scavenging assays, as well as the ferric reducing antioxidant power assay. Cellular antioxidant activity was determined by superoxide and nitric oxide scavenging assays. The cytotoxicity of the leaf extracts in RAW 264.7 and differentiated HL-60 cells was tested in parallel using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assays, respectively. The total phenolic and flavonoid contents were also assessed by spectrophotometric analysis. Phytochemical constituents of the most potent extract were investigated by liquid chromatography with an electrospray ionization quadrupole time-of-flight mass spectrometer (LC-ESI-QTOF-MS/MS). RESULTS: The ethanolic (ME) and decoction (LW) extracts of dried leaves had the highest chemical scavenging activity against DPPH and ABTS⁺ free radicals with half maximal effective concentration (EC50) values ranging from 3.54 to 6.44 µg/mL. ME and LW exerted moderate ferric reducing activity, with ferric reducing antioxidant power values of 847.41 and 941.26 mg Fe2+/g extract, respectively. Similarly, ME showed potent cellular scavenging activity against superoxide and nitric oxide radicals with EC50 values of 58.12 and 71.90 µg/mL, respectively. However, LW exhibited only strong nitric oxide scavenging activity with an EC50 value of 91.20 µg/mL. The cell viability of RAW 264.7 and HL-60 cells was greater than 70% in all tested concentrations of both extracts, thus confirming the absence of their cytotoxicity. ME and LW contained high total phenolic contents of 231.14 and 274.42 mg gallic acid equivalents per gram, respectively, as well as high total flavonoid contents of 18.82 and 22.17 mg quercetin equivalents per gram, respectively. LC-ESI-QTOF-MS/MS analysis revealed the presence of 52 structurally characterized compounds in ME, 43 of which were tentatively identified. Hydroxycinnamic acids such as caffeic acid and its derivatives were the predominant phenolic compounds. CONCLUSION: This is the first report describing potent chemical and cellular antioxidant effects of the ethanolic leaf extract of A. thwaitesianum. The extract contained high total phenolic and flavonoid contents. LC-ESI-QTOF-MS/MS analysis further revealed an abundance of caffeic acid derivatives and flavonoids. These data support its potential use as dietary supplements in oxidative stress prevention.

Cytotoxic, Anti-inflammatory and Antioxidant Activities of Heliotropium indicum Extracts.

Background: Heliotropium indicum Linn., or 'Indian heliotrope' is very common in India with a long history of traditional medicinal uses in many countries in the world. In Thailand, the plant has been traditionally use to cure various diseases such as fever, insect bites, stings, diarrhea, skin rashes, menstrual disorder and urticaria. In addition, the plant is commonly used by Thai folk doctors as a component in remedies for treatment of lung cancer. Objective: In the present study, we investigated cytotoxicity against two types of lung cancer cell lines (A549 and NCI-H226), anti-inflammatory effect and antioxidant activity of Heliotropium indicum extracts. Material and Method: The water and ethanolic extracts of Heliotropium indicum were tested. The cytotoxic activity against two types of human lung cancer cell lines (A549 and NCI-H226) was evaluated by sulforhodamine B (SRB) assay. The antiinflammatory effect was investigated on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. LPS-induced nitric oxide (NO) production was determined by Griess reagent. The antioxidant activity was performed by 1, 1-diphenyl-picrylhydrazyl (DPPH) radical scavenging method. Results: The ethanolic extract showed cytotoxic activity only against NCI-H226 (IC50 = 51.90±2.35 µg/ml) whereas the water extract had no cytotoxic activity against both A549 and NCI-H226 (IC50 >100 µg/ml). For anti-inflammatory effect, the results revealed that the ethanolic extract exhibited the most potent inhibitory activity on nitric oxide production (IC50 = 24.17±2.12 µg/ml), followed by Indomethacin (positive control) with an IC50 value of 34.67±6.23 µg/ml while water extract was apparently inactive (IC50 >100 µg/ml). For antioxidant activity, the ethanolic extract showed high antioxidant activity (EC50 = 28.91±4.26 µg/ml) but the water extract showed no antioxidant activity (EC50 >100 µg/ml). Conclusion: These results can support using Heliotropium indicum Linn. for component in lung cancer remedy by Thai folk doctors. However, more studies are required.

Development and validation of RP-HPLC method to determine anti-allergic compound in Thai traditional remedy called Benjalokawichien.

Benjalokawichien (BLW) or Ya-Ha-Rak (HR) is a traditional remedy in the Nationaldrug list of herbal medicinal products AD 2012 of Thailand. For traditional use, BLW is used as antipyretic agent. It also has anti-allergic effect, particularly treating allergic rash. The ethanolic extract of BLW exhibited anti-allergic activity via inhibitory effect against a release ofbeta-hexosaminidase in RBL-2H3 cell line. Pectolinarigenin has been identified as the active compound ofBLW extract. In this study, a reversed-phase high performance liquid chromatography (RP-HPLC) method was developed in order to control quality ofpreparation in three aspects such as chemical fingerprint, quantification and stability of the ethanolic extract. The RP-HPLC was performed with a gradient mobile phase composed of 0.1% ortho phosphoric acid and acetronitrile, and peaks were detected at 331 nm. Based on validation results, this analytical method is precise, accurate and stable for quantitative determination ofpectolinarigenin. The amount ofpectolinarigenin in Benjalokawichien extract determined by this method was 18.50 mg/g ofextract. Therefore, this method could be consideredfor quality control ofBLWextract.

Anti-allergic, anti-inflammatory and antioxidant activities of the different extracts of Thai traditional remedy called prabchompoothaweep for allergic rhinitis treatment.

BACKGROUND: Prabchompoothaweep remedy (PT) has long been used in Thai traditional medicine to treat allergic rhinitis and asthma. It is composed of 23 plants. It is on National herbal drug list of Thailand, but there is no reportfor anti-allergic, anti-inflammatory and antioxidant activities. OBJECTIVE: To investigate anti-allergic, anti-inflammatory and antioxidant activities of the crude extract from PTby different extraction method. MATERIAL AND METHOD: The method of extract used was maceration in 95% ethanol and 50% ethanol; the residue of these extracts were continued extracted by boiling water, they obtained PTE95, PTE50, PTR95 andPTR50, respectively. The other method of extraction was boiling and drying by lyophilizer that obtained PTW Five crude extracts were determined anti- allergic activity by the inhibition of ß-hexosaminidase release from RBL-2H3 cell lines, anti-inflammatory activity were determined by the inhibition ofnitric oxide (NO) production from RA W264. 7 cell lines induced by lipopolysaccharide (LPS) and antioxidant activity were tested by DPPH radical scavenging assay. RESULTS: PTE95 showed the most potent ofanti-allergic activity, anti-inflammatory activity and antioxidant activity (IC5 = 12.97, 22.51 and EC50 = 14.62 µg/ml, respectively). CONCLUSION: These results suggest that the method of extraction PT that showed the best anti-allergy, anti-inflamation and antioxidant activity was maceration in 95% ethanol.

Inhibitory Effect on ß -Hexosaminidase Release from RBL-2H3 Cells of Extracts and Some Pure Constituents of Benchalokawichian, a Thai Herbal Remedy, Used for Allergic Disorders.

Introduction. Benchalokawichian (BCW), a Thai traditional herbal formulation, has long been used as antipyretic and to treat skin disorders. It comprises roots from five herbs: Ficus racemosa, Capparis micracantha, Clerodendrum petasites, Harrisonia perforata, and Tiliacora triandra. This polyherbal remedy has recently been included in the Thailand National List of Essential Medicines (Herbal Products list). Methodology. A Bioassay-guided fractionation technique was used to evaluate antiallergy activities of crude extracts, and those obtained by the multistep column chromatography isolation of pure compounds. Inhibitory effect on the release of ß-hexosaminidase from RBL-2H3 cells was used to determine antiallergic activity. Results. Two pure compounds from BCW formulation showed higher antiallergic activity than crude or semipure extracts. Pectolinarigenin showed the highest antiallergic activity, followed by O-methylalloptaeroxylin, with IC50 values of 6.3 µg/mL and 14.16 µg/mL, respectively. Moreover, the highest activities of pure compounds were significantly higher than chlorpheniramine (16.2 µg/mL). Conclusions. This study provides some support for the use of BCW in reducing itching and treatment of other skin allergic disorders. The two isolated constituents exhibited high antiallergic activity and it is necessary to determine their mechanism of action. Further phytochemical and safety studies of pure compounds are required before development of these as antiallergy commercial remedies.