Effects of the Thai Herbal Wattana Formula and Its Ingredients in the Human Hepatocarcinoma HepG2 Cells: Safety and Efficacy Considerations

Booranasubkajorn, S orcid iconORCID: 0000-0002-3192-878X, Lumlerdkij, N orcid iconORCID: 0000-0001-5142-8507, Chaisri, SMATM orcid iconORCID: 0000-0001-7433-2110, Akarasereenont, P and Prieto Garcia, JM orcid iconORCID: 0000-0002-2649-1691 (2025) Effects of the Thai Herbal Wattana Formula and Its Ingredients in the Human Hepatocarcinoma HepG2 Cells: Safety and Efficacy Considerations. Natural Product Communications, 20 (10). ISSN 1934-578X

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Abstract

Introduction: The Thai Herbal Wattana formula (WNF) is a multi-ingredient remedy used to promote overall health and mitigate age-related physiological degeneration, suggesting a potential adjuvant use in oncological treatments. Aims: This ethnopharmacological study aimed to evaluate cytotoxic and antimigratory effects of the Ayurveda Siriraj WNF AVS073 variation (ASW) and its active/s botanical constituents in human liver cancer (HepG2) cells. Methods: ASW and its ingredients were evaluated for cytotoxicity (Alamar blue), anti-migratory activity (2D gap closure). Mechanistic studies included cell death, apoptosis and cell cycle arrest (flow cytometry), and intracellular glutathione levels. Bioguided isolation was used to identify the active compound/s. Results: ASW did not inhibit HepG2 cell proliferation at 200 µg/mL although it halved intracellular glutathione levels and reduced cell migration similarly to paclitaxel 0.01 nM. In contrast, the water extract from Biancaea sappan (syn. Caesalpinia sappan L.) (CSL) was the only ingredient showing cytotoxicity (IC50=44 µg/ml). It induces apoptosis and G2/M phase cell cycle arrest and significantly reduced 2D cell migration without modifying glutathione levels. Brazilein was dereplicated as the active cytotoxic component in CSL but it did not show any effect on glutathione levels or 2D cell migration. Conclusion: This preclinical study demonstrates that the ASW formula lacks direct in vitro cytotoxicity towards HepG2 cells, but effectively reduced cell mobility and intracellular glutathione although at non-physiological concentrations. B. sappan L., exhibits potent cytotoxic and anti-migratory activities. Brazilein is its primary cytotoxic compound although is not endowed with the glutathione-depleting or anti-migratory effects observed in the ASW formula. These findings suggest that ASW’s benefits in oncology may primarily link up with its established immunological and anti-inflammatory effects, while its lack of toxicity to HepG2 cells, a proxy for hepatocytes, plus its clinically proven lack of major adverse effects might indicate a positive safety profile

Item Type: Article
Uncontrolled Keywords: 0305 Organic Chemistry; 0602 Ecology; 0607 Plant Biology; 3214 Pharmacology and pharmaceutical sciences; 3405 Organic chemistry
Subjects: R Medicine > RS Pharmacy and materia medica
Divisions: Pharmacy and Biomolecular Sciences
Publisher: SAGE Publications
Date of acceptance: 19 September 2025
Date of first compliant Open Access: 8 October 2025
Date Deposited: 08 Oct 2025 10:31
Last Modified: 08 Oct 2025 10:45
DOI or ID number: 10.1177/1934578x251385040
URI: https://researchonline.ljmu.ac.uk/id/eprint/27295
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