Aqueous Extracts of Liver Kidney CareTM and GingerTM Inhibit Intracellular Inflammatory Signaling Cascade

Abstract The study aims to assess the in vitro anti-inflammatory property of aqueous extracts of novel herbal formulations consisting of Phyllanthus niruri, Boerhavia diffusa and Picrorhiza kurroa (Liver Kidney CareTM– LKCTM,Organic IndiaTM) and Zingiber officinale (Ginger, Organic IndiaTM). The extracts were analyzed by Liquid Chromatography-Mass Spectrometry (LC-MS) for the detection of active ingredients. LC-MS confirmed the presence of active ingredients Phyllanthin, Boeravinone-B, Picroside I and Picroside II in LKCTM; and 6-GingerolinGinger. Cytotoxicity assessments were done by Trypan blue cell exclusion method. The cells were pre-incubated with extracts from two herbal formulations LKCTMor Ginger prior to TNFαor arachidonic acid (AA) stimulation to determine their effects on inflammatory signalling. Non-toxic doses of the extracts were used to determine their anti-inflammatory property by quantifying mRNA expression of IL-6, IKBA, ALOX5 and COX2 using qPCR and protein expression of IκBα and phospho-p65 by western blot.TNFα-induced mRNA expression of IL-6, IKBA and COX2, and IκBα protein degradation along with phosphorylation of p65 was inhibited by LKCTM and Ginger extracts. AA-induced mRNA expression of COX2, ALOX5 and IL-6, and phosphorylation of p65 was inhibited by LKCTM and Ginger extracts.LKCTM and Ginger aqueous extracts inhibited TNFα- or AA-induced intracellular inflammatory signalling pathways (NFκB, cyclooxygenase, lipooxygenase),suggesting the anti-inflammatory potential of these herbal formulations.