Context: Chatuphalatika (CTPT), is a Thai herbal formulation mixture of Phyllanthus emblica Linn. (Euphorbiaceae), Terminalia belerica Linn. (Combretaceae), T. chebula and the fruit of T. arjuna (Roxb.) Wight & Arn. CTPT is considered to exert anti-inflammatory and antihyperuricemic effects, but there have been no reports to demonstrate these pharmacological effects in a quantitative manner. Objectives: To investigate the antioxidative, anti-inflammatory and antihyperuricemic effects of CTPT. Materials and methods: Antioxidant activities of CTPT extracts were measured in vitro by DPPH, ABTS and FRAP assays, and anti-inflammatory effect by measuring inflammatory mediator production induced by lipopolysaccharide (LPS) in RAW264.7 macrophages. The mechanism of the hypouricemic effect was investigated using oxonate-induced hyperuricemic ddY mice treated with oral administrations of CTPT at 250, 500 and 1000 mg/kg. Results: Antioxidant activities of CTPT measured by ABTS and FRAP assays were 1.35 g TEAC/g extract and 10.3 mmol/100 g extract, respectively. IC50 for the inhibition of DPPH radical was 13.8 µg/mL. CTPT (10 µg/mL) significantly downregulated the mRNA expression of TNF-α and iNOS in RAW 264.7 cells. Lineweaver–Burk analysis of the enzyme kinetics showed that CTPT inhibited xanthine oxidase (XOD) activity in a noncompetitive manner with the Ki of 576.9 µg/mL. Oral administration of CTPT (1000 mg/kg) significantly suppressed uric acid production by inhibiting hepatic XOD activity, and decreased plasma uric acid levels in hyperuricemic mice by approximately 40% (p < 0.05). Conclusions: This study demonstrated for the first time the antioxidative, anti-inflammatory and antihyperuricemic effects of CTPT in vivo and in vitro, suggesting a possibility of using CTPT for the treatment of hyperuricemia in gout.

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The objectives of the present study were to demonstrate the antihyperuricemic effect of triphala, a formulation of traditional Ayurvedic medicine, in potassium oxonate-induced hyperuricemic mice in vivo, and to examine its inhibitor effects on xanthine oxidase (XOD), inflammatory mediators and DPPH radicals in vitro. The water extract of triphala was determined to contain the total phenolics and total flavonoids of 317.6 ± 9.2 mg GAE/g and 7.73 ± 0.26 mg QE/g, respectively. Oral administrations of triphala significantly reduced the plasma uric acid levels of potassium oxonate-induced hyperuricemic mice at doses of 1,000 and 1,500 mg/kg, as compared with control ( p<0.05). Moreover, both doses of triphala treatments markedly inhibited the formation of uric acid due to inhibition of XOD activity in liver homogenates extracted from the hyperuricemic mice by about 70-80%. Lineweaver-Burk analysis of enzyme-kinetic data showed that triphala exhibited non-competitive inhibition on XOD activity in vitro with an inhibitory constant (Ki) of 590 μg/mL. Furthermore, triphala significantly suppressed the mRNA expressions of COX-II, TNF-α and iNOS in LPS-stimulated RAW 264.7 cells, as compared with control ( p <0.05), and decreased the expression of TGF-β. IC50 values for inhibition of DPPH radical formation was calculated to be 21.9 ± 2.50 μg/mL. Antioxidant activities of triphala were determined to be 0.81 ± 0.07 g TEAC/g and 6.78 ± 0.29 mmol/100g, respectively, as assessed by ABTS and FRAP assays. In conclusion, this study provided in vivo and in vitro mechanistic evidence for the antihyperuricemic, antioxidative and anti-inflammatory effects of triphala for the first time, rationalizing its therapeutic usage for the treatment of hyperuricemia of gout.

Croton stellatopilosus Ohba (Plau-noi), a well-known Thai medicinal plant, was investigated for its genetic variation by analyzing three DNA regions, one nuclear internal transcribed spacer (ITS) region and two chloroplast trnL-F intergenic spacer and trnK intron regions. The results of ITS sequencing from 30 leaf samples showed that there were two major genotypes of C. stellatopilosus which were designated as STEL Type A and B. In addition, various nucleotide additive sequences which had presumably arisen from these two groups were also found. These so-called "putative hybrids", interestingly, displayed trnK intron sequences identical to the STEL Type B but different from the Type A. For the trnL-F region, all the 30 samples showed identical sequences. Thus, it was suggested that in the hybridization of C. stellatopilosus, the Type A genotype acts as paternal parent whereas the Type B genotype acts as maternal parent. In addition, all C. stellatopilosus samples were analyzed for their plaunotol content using TLC densitometry. We found that the Type A genotype, hybrid group and Type B genotype had plaunotol content in the ranges 0.209-0.492, 0.319-0.896 and 0.442-1.000% (w/w) dry weight, respectively. The results indicated that there is a correlation between the plaunotol contents and non-coding DNA sequences of ITS, trnK and trnL-F regions of C. stellatopilosus.

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