Garawadmath, S. | bioRxiv Science

bioRxiv2023

Multicomponent Ayurveda formulation Lodhrasavam ameliorates steatosis and lipotoxicity in HepG2 cell model of NAFLD

BackgroundMetabolic-dysfunction associated steatotic liver disease (MASLD) is a complex, multifactorial condition and a leading cause of liver-related morbidity worldwide. Due to its heterogeneous pathogenesis, conventional single-target therapies often show limited efficacy. Multicomponent ayurveda formulations offer a promising alternative through their multitargeted actions. This study explores the therapeutic potential and underlying mechanisms of Lodhrasavam (LS), a classical, polyherbal ayurveda formulation, in both in vitro and in vivo models of MASLD. MethodsLS was evaluated for its anti-steatotic and anti-obesogenic potential using in vitro (HepG2, 3T3-L1) and in vivo (HF-HFD-induced rat) models. Antioxidant, lipase inhibition, cytotoxicity, anti-steatotic, and anti-adipogenic activities were assessed via DPPH, MTT, ORO/BODIPY staining, triglyceride quantification, ROS assay, and qPCR. Lipidomic profiling was done in HepG2 cells. Biochemical analyses (GLP-1, insulin, lipid panel), histopathology and OGTT further validated efficacy. Data were statistically analyzed by one-way ANOVA (p<0.05). Results/discussion: In HepG2 cells, LS reduced PA-induced lipid accumulation, ROS, triglycerides, and restored viability, while downregulating lipogenic genes (PPAR{gamma}, SREBP-1c, FASN). Lipidomics confirmed lipid modulation. In 3T3-L1 cells, LS suppressed adipogenesis and key adipogenic genes. In HFD-fed rats, LS reduced weight gain, hepatic steatosis, serum lipids, and improved liver histology. LS enhanced insulin and GLP-1 secretion, improved glucose tolerance, and restored pancreatic islet structure. These findings highlight the multitargeted potential of LS in ameliorating MASLD by modulating lipid metabolism, oxidative stress, inflammation, and glucose homeostasis. ConclusionLodhrasavam ameliorates MASLD by modulating the liver-adipose-pancreas axis, improving lipid metabolism, reducing inflammation, and enhancing insulin and GLP-1 secretion through multitargeted mechanisms.

pharmacology and toxicology↗

bioRxiv2022

In vitro and in silico analysis proving DPP4 inhibition and diabetes associated gene network modulation by a polyherbal formulation - Nisakathakadi Kashaya.

Frontiers of disease biology started recognizing the importance of systems and network medicine approach for managing chronic disease like diabetes. Dipeptidyl-peptidase IV (DPP4) inhibitors are one such class of anti-diabetic drugs recognized for their systemic biological actions. Polyherbal preparations like Ayurveda formulations are ideal for identifying novel DPP4 inhibitors having greater efficacy and safety profile. Additionally, expanding the research on the multitargeted mode of action of these polyherbal formulations can render novel insights into the complex biology of disease manifestations. The current study aims at identifying DPP4 inhibitory potential of a clinically established Ayurveda anti-diabetic formulation Nisakathakadi Kashaya (NK) using in vitro and in silico methods as well as the modulation of diabetes associated gene network by NK. a. Standard enzyme inhibition assay was used to study the DPP4 inhibitory potential of NK, followed by bioinformatics and computational biology tools for identifying the potential bioactives and their molecular interactions involved in DPP4 inhibition. STITCH, CHEMBL and BindingDB databases were used for target mapping and depicting the multi-targeted network pharmacology interaction of NK and the formulation. EnrichR was used to depict a sub-network of diabetes proteins and their relationship with diabetes associated comorbidities. NK demonstrated a dose dependent DPP4 inhibition with an IC50 of 2.06 g GAE/mL. Molecular docking identified three compounds namely Terchebin, Locaracemoside B and 1,2,4,6 Tetra o Galloyl Beta D Glucose showing stable interactions with DPP4 similar to the standard drug Vildagliptin. The network pharmacology analysis of NK identified a number of targets like TNF, TGF{beta}1, SOD1, SOD2, AKT1, DPP4 and GLP1R in its protein-protein interaction network which are vital to diabetic progression and complications. The present work demonstrated that the polyherbal formulation NK has DPP4 inhibition potential and modulates a large number of diabetes related proteins and pathways. The approach adopted in the current study by combining in vitro and in silico methods allowed us to understand the mechanism of DPP4 inhibition by the formulation and also the possible pharmacological networking through which the formulation exert its systemic effect in diabetes management.

systems biology↗