Individual Botanicals Perform Better than AYUSH-64 Formulation in Suppressing TLR4-Driven Inflammatory Cytokines

Area of Science:

• Immunology and Pharmacology
• Natural Product Chemistry

Background:

• Dysregulated macrophage activation via Toll-like receptor 4 (TLR4) signaling is implicated in inflammatory diseases.
• AYUSH-64, a polyherbal formulation, is being investigated for its immunomodulatory properties.
• The specific anti-inflammatory contributions of AYUSH-64's individual botanicals (Alstonia scholaris, Picrorhiza kurroa, Swertia chirata, Caesalpinia crista) are not well understood.

Purpose of the Study:

• To compare the prophylactic immunomodulatory effects of AYUSH-64 and its individual extracts on lipopolysaccharide (LPS)-induced inflammation in macrophage-like cells.
• To investigate the impact of these compounds on Toll-like receptor 4 (TLR4) signaling pathways.

Main Methods:

• THP-1 monocytes were differentiated and stimulated with LPS to activate TLR4.
• Cells were pre-treated with AYUSH-64 or individual extracts (Alstonia scholaris, Picrorhiza kurroa, Swertia chirata, Caesalpinia crista) or dexamethasone (positive control).
• Gene expression and protein secretion of pro-inflammatory (TNF-α, IL-1β) and regulatory (IL-10) cytokines were quantified using qRT-PCR and ELISA.

Main Results:

• AYUSH-64 demonstrated moderate suppression of TNF-α and IL-10, with minimal effect on IL-1β.
• Swertia chirata (SC), Picrorhiza kurroa (PK), and Caesalpinia crista (CC) extracts showed more pronounced and selective inhibition of inflammatory markers.
• SC and PK significantly reduced IL-10 secretion, surpassing dexamethasone's efficacy; CC exhibited dose-dependent IL-10 reduction.

Conclusions:

• Individual components of AYUSH-64, especially SC, PK, and CC, possess stronger and more selective in vitro anti-inflammatory properties than the whole formulation.
• These botanicals, particularly SC and PK, are more effective in reducing IL-10 secretion.
• Further in vivo and clinical studies are necessary to validate the therapeutic potential of these individual plant extracts.