Safety Evaluation of the Polyherbal Formulation NawaTab: Acute and Subacute Oral Toxicity Studies in Rats.

NawaTab is a tablet formulation developed from the Nawametho polyherbal formula used in Surat Thani province, Southern Thailand, for the treatment of hyperlipidemia. This study aims at evaluating the acute and subacute toxicity of NawaTab in rats. In the acute toxicity study, NawaTab was evaluated in female rats following the OECD Guideline No. 423. In the subacute toxicity study, NawaTab was tested in both male and female rats following the OECD Guideline No. 407. In the acute toxicity study, no lethal effects or toxic signs were observed during the duration of the study. In the subacute toxicity study, there was no mortality and no abnormality in clinical signs, body weight, food consumption, relative organ weight, and hematological parameters of NawaTab-treated rats. Significantly increased water consumption by male rats (500 mg/kg BW) and female rats (250, 500, and 1000 mg/kg BW) was observed. In addition, globulin and total cholesterol of female rats (1000 mg/kg BW) significantly increased. These alterations were within normal physiological ranges. Moreover, necropsy and histopathological findings of NawaTab-treated rats demonstrated no obvious alterations attributable to NawaTab administration. The present study revealed that NawaTab has no significant acute oral toxicological effects. The lethal dose with a 50% mortality rate (LD50) was higher than 5000 mg/kg BW in rats. The subacute oral administration of NawaTab for 28 days did not have any major toxicological effects. Based on this study, NawaTab could be safe to use with caution pending its chronic toxicity study.

Supplementation with the Traditional Thai Polyherbal Medicine NawaTab Ameliorates Lipid Profiles in High-Fat Diet-Induced Hyperlipidemic Rats.

A tablet formulation developed from Nawametho decoction (NawaTab), a traditional Thai herbal mixture described in the Worayokasan scripture, has been used clinically for several years in the management of borderline hyperlipidemic individuals. Nevertheless, scientific evidence supporting its claims has never been identified. This study aimed to describe the antihyperlipidemic properties of NawaTab in a well-described model of high-fat diet (HFD)-induced hyperlipidemic rats. Decoction of Nawametho containing equal quantities of Aegle marmelos (L.), Carthamus tinctorius L., Hibiscus sabdariffa Linn., Phyllanthus emblica L., Piper longum L., Piper nigrum L., Terminalia bellirica (Gaertn.) Roxb., Terminalia chebula Retz., and Zingiber officinale Roscoe were prepared. The HFD-fed rats were administered NawaTab for 4 consecutive weeks starting from the ninth week of HFD treatment at a dose of 125 mg/kg (p.o.). Anthropometric parameters, serum lipid profiles, lipase activity, and liver and renal functional markers were measured. Histopathological examination of the liver and any steatosis was carried out at the end of this study. Consecutive treatment with NawaTab (125 mg/kg/day) in HFD-induced hyperlipidemic rats caused a significant reduction in serum total cholesterol, triglyceride, low-density lipoprotein cholesterol, and very low-density lipoprotein cholesterol levels. However, at the tested dose, NawaTab failed to prevent the onset of hepatic steatosis and adipose tissue accumulation. No adverse events due to the consumption of NawaTab on liver and kidney function markers were noted. These findings are the first suggestive evidence for the lipid-lowering capability of NawaTab. However, to promote the use of this formulation in the management of borderline hyperlipidemic patients, elucidation of the underlying mechanisms of action, quantification of biological markers, and clinical trials of NawaTab are urgently needed.

The Standardized Wood Extract of Albizia myriophylla: Its Potential as an Active Ingredient in an Anti-inflammatory Herbal Gel Formulation.

Albizia myriophylla has been used in Thai folk medicine for treating inflammation-related diseases. The wood of this medicinal plant is traditionally used as a single herbal drug in the form of an aqueous decoction and as a component in several Thai herbal formulations for the remedy of fever, sore throat, and aphthous ulcers. This study aimed to evaluate in vivo the anti-inflammatory potential and possible mechanism of action of the standardized wood extract of A. myriophylla as well as to investigate the anti-inflammatory activity and physicochemical properties of the developed herbal gel formulation containing standardized wood extract of A. myriophylla. Results of quantitative HPLC analysis demonstrated that the standardized wood extract of A. myriophylla contained 22.95 mg/g of 8-methoxy-7,3',4'-trihydroxyflavone, a bioactive marker compound of A. myriophylla. The standardized wood extract of A. myriophylla (1% w/v) exhibited remarkable inhibition (54.4 - 80.3%) in the croton oil model of topical inflammation at all assessment times, comparable to standard indomethacin (55.3 - 63.6%). Real-time quantitative reverse transcription-polymerase chain reaction was performed to clarify the anti-inflammatory mechanism of standardized wood extract of A. myriophylla, and the result showed that this standardized extract decreased lipopolysaccharide-induced nitric oxide synthase mRNA levels in a dose-dependent manner. The developed herbal gel containing standardized wood extract of A. myriophylla (1% w/w) had good physicochemical characteristics and exhibited potent inhibition (51.4 - 77.8%) of inflammation in a rat ear edema model at all assessment times, comparable to indomethacin gel (33.3 - 40.5%). The notable anti-inflammatory activity of standardized wood extract of A. myriophylla and its developed herbal gel formulation indicates their potential application as natural anti-inflammatory agents.

Traditional tonifying polyherbal infusion, Jatu-Phala-Tiga, exerts antioxidant activities and extends lifespan of Caenorhabditis elegans.

BACKGROUND: The imbalance between the generation of free radicals and natural cellular antioxidant defenses, known as oxidative stress, can cause oxidation of biomolecules and further contribute to aging-associated diseases. The purpose of this study was to evaluate the antioxidant capacities of Thai traditional tonifying preparation, Jatu-Phala-Tiga (JPT) and its herbal ingredients consisting of Phyllanthus emblica, Terminalia arjuna, Terminalia chebula, and Terminalia bellirica and further assess its effect on longevity. METHOD: Antioxidant activities of various extracts obtained from JPT and its herbal components were carried out using well-established methods including metal chelating, free radical scavenging, and ferric reducing antioxidant power assays. Qualitative analysis of the chemical composition from JPT water extract was done by high-performance liquid chromatography tandem with electrospray ionisation mass spectrometry. The effect of JPT water extract on the lifespan of Caenorhabditis elegans were additionally described. RESULTS: Among the extracts, JPT water extract exerted remarkable antioxidant activities as compared to the extracts from other solvents and individual constituting plant extract. JPT water extract was found to possess the highest metal chelating activity, with an IC50 value of 1.75 ± 0.05 mg/mL. Moreover, it exhibited remarkable scavenging activities towards DPPH, ABTS, and superoxide anion radicals, with IC50 values of 0.31 ± 0.02, 0.308 ± 0.004, and 0.055 ± 0.002 mg/mL, respectively. The ORAC and FRAP values of JPT water extract were 40.338 ± 2.273 µM of Trolox/µg of extract and 23.07 ± 1.84 mM FeSO4/mg sample, respectively. Several well-known antioxidant-related compounds including amaronols, quinic acid, gallic acid, fertaric acid, kurigalin, amlaic acid, isoterchebin, chebulagic acid, ginkgolide C, chebulinic acid, ellagic acid, and rutin were found in this extract. Treatment with JPT water extract at 1 and 5 mg/mL increased C. elegans lifespan under normal growth condition (7.26 ± 0.65 vs. 10.4 0± 0.75 (p < 0.01) and 10.00 ± 0.73 (p < 0.01) days, respectively). CONCLUSIONS: The results indicated that JPT and its herbal ingredients exhibited strong antioxidant activities, in particular the water extract of the polyherbal tonic. These findings rationalize further investigation in JPT infusion as a promising agent for anti-aging and oxidative stress prevention.

Effects of a traditional Thai polyherbal medicine 'Ya-Samarn-Phlae' as a natural anti-biofilm agent against Pseudomonas aeruginosa.

Ya-Samarn-Phlae (YaSP) is a traditional Thai polyherbal formula for the treatment of chronic wounds. Although its ethanol extract has been proven to possess several wound-related biological activities, there is no scientific information available for the infused oil of YaSP, which is its traditionally prepared form. This present study therefore aimed to evaluate the efficacy of different infused oils obtained from either fresh or oven-dried herbal parts of YaSP (F-YaSP and D-YaSP) against biofilms of Pseudomonas aeruginosa, which reside in chronic wounds. Its main active herbal component, Garcinia mangostana (F-GM and D-GM), as well as α-mangostin were also tested in this study. All infused oils significantly inhibited the biofilm formation of P. aeruginosa with a percentage of reduction ranging from 50 to 90%. Visualization of the inhibition of biofilm development was confirmed using scanning electron and atomic force microscopes. All tested agents resulted in a reduction in the mean average roughness of the biofilm, whereas only treating with D-YaSP, D-GM, and α-mangostin led to a decrease in both peak height and peak-valley height. MTT reduction assays revealed that the metabolic activity of P. aeruginosa mature biofilms decreased considerably up to 50% after only 3 h of incubation and after only 9 h of exposure to D-YaSP. Confocal laser scanning micrographs illustrated that a maximum biofilm eradication was found when treated with the extracts for 3 h, whereas the biomass, the average thickness, maximum thickness, and the surface to volume ratio of the treated biofilm was reduced after up to 18 h of contact time. It can be concluded that D-YaSP can effectively inhibit biofilm formation and eradicate mature biofilms of P. aeruginosa. It should be noted that G. mangostana and α-mangostin contribute in YaSP as principle active agents for anti-biofilm efficacy.

Bioactive components, antioxidant, and anti-inflammatory activities of the wood of Albizia myriophylla

Abstract The scientific basis corresponding with the folkloric use of Albizia myriophylla Benth., Fabaceae, for the treatment of inflammation-related diseases was established by measuring antioxidant potential using 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) free radicals, and ferric reducing antioxidant power assays as well as anti-inflammatory effect using nitrite assay and ethyl phenylpropiolate (EPP)-induced rat ear edema model. Both ethanol extract (DPPH, IC50 46.23 µg/ml; ABTS, IC50 57.14 µg/ml; FRAP, 950.14 mM Fe (II)/g) and dichloromethane fraction (DPPH, IC50 29.54 µg/ml; ABTS, IC50 40.36 µg/ml; FRAP, 946.69 mM Fe (II)/g) from A. myriophylla demonstrated a promising antioxidant activity. Furthermore, it was found that the ethanol extract of A. myriophylla showed significant inhibitory activity against lipopolysaccharide (LPS)-induced nitric oxide production in murine macrophage cells (IC50 13.8 µg/ml). The ethanol extract (15% w/v) also exhibited the maximum percentage inhibition (81-95%) of inflammation in the ear edema model at all assessment times comparable to indomethacin (0.5 mg/ear). Among all isolates 1-5 from the active extract of A. myriophylla, indenoic acid (1) (DPPH, IC50 8.96 µg/ml; ABTS, IC50 10.12 µg/ml) and 8-methoxy-7,3′,4′-trihydroxyflavone (2) (DPPH, IC50 5.05 µg/ml; ABTS, IC50 7.89 µg/ml) had potent free radical scavenging effects comparable to those of ascorbic acid (DPPH, IC50 2.12 µg/ml; ABTS, IC50 3.26 µg/ml). Compound 2 also displayed remarkable reducing power in FRAP test (261.81 mg QE/g) and showed a marked inhibition of the cellular nitric oxide production (IC50 27.7 µg/ml). Our results suggest that the anti-inflammatory mechanism of A. myriophylla is most probably based on its capacity to suppress nitric oxide production as well as to be free radical scavenger.

Antioxidant capacities and total phenolic contents of 20 polyherbal remedies used as tonics by folk healers in Phatthalung and Songkhla provinces, Thailand.

BACKGROUND: Uses of polyherbal formulations have played a major role in traditional medicine. The present study is focused on the formulations used in traditional Thai folkloric medicine as tonics or bracers. Twenty documented polyherbal mixtures, used as nourishing tonics by the folk healers in Phatthalung and Songkhla provinces in southern Thailand, are targeted. Despite traditional health claims, there is no scientific evidence to support the utilization of polyherbal formulations. METHODS: The phenolic and flavonoid contents of the polyherbal formulations and a series of antioxidant tests were applied to measure their capability as preventive or chain-breaking antioxidants. In addition, the cytotoxic activity of effective formulations was assayed in Vero cells. RESULTS: Ninety-eight plant species belonging to 45 families were used to prepare the tested formulation. The preliminary results revealed that water extracts of THP-R016 and THP-R019 contain a high level of total phenolic and flavonoid contents and exhibit remarkable antioxidant activities, as tested by DPPH, ABTS, and FRAP assays. The extract of THP-R019 also showed the strongest metal chelating activities, whereas THP-R016 extract possessed notable superoxide anion and peroxyl radical scavenging abilities. CONCLUSIONS: The data provide evidence that the water extracts of folkloric polyherbal formulations, particularly THP-R016, are a potential source of natural antioxidants, which will be valuable in the pharmaceutical and nutraceutical industries. The free radical scavenging of THP-R016 may be due to the contribution of phenolic and flavonoid contents. Useful characteristics for the consumer, such as the phytochemical profiles of active ingredients, cellular based antioxidant properties and beneficial effects in vivo, are under further investigation.

Longan (Dimocarpus longan Lour.) inhibits lipopolysaccharide-stimulated nitric oxide production in macrophages by suppressing NF-κB and AP-1 signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Flower, seed, and fruit of longan (Dimocarpus longan Lour.) have been used in the Traditional Chinese Medicine (TCM) serving as a common herb in relief of swelling which can be applied in cases of inflammatory diseases. However, the scientific evidence related to their effects on inflammation especially the possible cellular and molecular mechanisms of longan need to be clarified. AIM OF THE STUDY: To evaluate the anti-inflammatory effect of the various parts of longan including flower, seed, and pulp. The mechanisms and molecular targets involved in their effects were also investigated. MATERIALS AND METHODS: Different longan extracts were analyzed for their bioactive compounds and evaluated for anti-inflammation. Corilagin, ellagic acid, and gallic acid were detected using HPLC-DAD. In vitro anti-inflammatory effect of longan extracts and their polysaccharides were examined by analyzing nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Expression of the proteins that involved in NO production was detected by Western blot. RESULTS: Flower extract contained the highest amounts of total phenolics, total flavonoids, proanthocyanidins, corilagin and ellagic acid when compared to seed and pulp extracts. The extracts of different longan parts inhibited LPS-induced NO production with different potency. The highest potency for the inhibition of NO production was shown with flower extract follow by seed and pulp (IC50=128.2, 1127.4, and 1260.2µgmL(-1), respectively). The mechanisms of this anti-NO production were associated with their NO scavenging effect and their decreasing the expression and catalytic activity of an inducible nitric oxide synthase (iNOS). Moreover, these longan extracts suppressed LPS-induced degradation of IκBα and activation of NF-κB, activator protein-1 (AP-1), Akt, and mitogen activated protein kinases (MAPKs). CONCLUSION: These results suggest that the longan extracts possess anti-inflammatory property. Therefore, longan could provide potential dietary supplement for the treatment of inflammatory-related diseases.