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.

Anti-Helicobacter pylori, Anti-Inflammatory, Cytotoxic, and Antioxidant Activities of Mace Extracts from Myristica fragrans.

The aril (mace) of Myristica fragrans, known as Dok-Chan, is a spice that has long been used for treating stomach discomfort, peptic ulcer, and nausea. It is an ingredient in many remedies in Thai traditional medicine, e.g., Ya-Hom-Thep-Bha-Jit, Ya-Hom-Nao-Wa-Kot, and Ya-That-Bun-Job, which are used to treat dyspepsia and other gastrointestinal tract symptoms. The aqueous and ethanolic extracts of mace were used for all tests. Anti-H. pylori activities were determined by the disc diffusion method and agar dilution. Anti-inflammatory activity was determined by the LPS-induced nitric oxide (NO) inhibition in a RAW264.7 cell line, and cytotoxicity was determined against gastric cancer cell lines (Kato III) using the sulphorhodamine B (SRB) assay. The DPPH radical scavenging and ABTS radical cation decolorization assays were used to determine the antioxidant activities. The result found that the ethanolic extract of mace exhibited antimicrobial activity against H. pylori ATCC 43504 and six clinical strains with MIC values of 125-250 µg/ml. The aqueous extract MICs against H. pylori ATCC reference strain and six clinical strains were 500 µg/ml compared with 0.5 µg/ml for the positive control, clarithromycin. The inhibitory effect of LPS-induced NO release and cytotoxic activity of the ethanolic extract had IC50 values of 82.19 µg/ml and 26.06 µg/ml, respectively, and the EC50 values for the DPPH and ABTS antioxidant assays were 13.41 µg/ml and 12.44 µg/ml, respectively. The mace extract also had anticancer properties. In conclusion, the ethanolic mace extract had anti-H. pylori, anti-inflammatory, antioxidant, and anticancer activities. These data support further preclinical and clinical investigation to see if the mace extract could have a role in treating patients with dyspepsia, peptic ulcers, and possibly gastric cancer.

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.

Comparison of antimicrobial, antioxidant activities and total phenolic content of Antidesma thwaitesianum fruit extracts by different methods.

BACKGROUND: Antidesma thwaitesianum Mull. Arg. is a tropical fruit in Northeast Thailand and its fruits are used for soft drink and healthy food. The biological activities of extracts from Antidesma thwaitesianum using different extraction methods have not been reported. OBJECTIVE: Extracts from Antidesma thwaitesianum using different extraction methods were tested for antibacterial and antioxidant activities as well as were determined for total phenolic content. Ten extracts were tested for antimicrobial activity by disc diffusion, MIC, and MBC methods. DPPH assay was used to test antioxidant activity and Folin-Ciocalteu 's reagent was used to determine total phenolic content. RESULTS: The extract obtained by decocting residue after maceration process of dried marc (MRW) exhibited stronger antioxidative power (EC50 = 11.73 microg/ml) than BHT (EC50 = 13.36 microg/ml). This antioxidant activity was related to total phenolic content of 85.77 +/- 0.34 mg GAE/g. This extract also exhibited antimicrobial activity against Staphylococcus aureus (inhibition zone = 8 mm, MIC = 2.5 mg/ml). Moreover the extract obtained by macerating dried marc (MME) exhibited antimicrobial activity against Bacillus subtilis with the same MIC value of 10 mg/ml. CONCLUSION: All of the extracts of Antidesma thwaitesianum had less potential for antimicrobial activity than Gentamicin and Amphotericin B. On the other hand, the water extract especially obtained by decocting residue after maceration of dry marc had good antioxidant power and the highest total phenolic content. Thus, such water extract should be recommended for a good source of natural antioxidants for commercial uses.