Disrupting Fibrogenic Pathways: Insilico and Invitro Evaluation of Kaempferol and Coumestrol as Potential Anti-Fibrotic Agents

Background:Liver fibrosis is caused by chronic liver injury which is a reversible process of excessive extracellular matrix deposition. The continuous activation of hepatic stellate cells progresses to fibrosis, resulting in cirrhosis and hepatocellular carcinoma. Conventional treatment focuses only on symptoms, but natural phytochemicals as antifibrotic drugs offer a prospective option for novel therapeutic techniques that target fibrogenic pathways.Objective: To explore the hepatoprotective potential of selected phytochemicals using computational and experimental approaches involving fibrogenic pathways.Methodology:Bioactive compounds of medicinal plants Euphorbia hirta(whole plant) and Lepidium sativum(seeds) were studied. The 69 phytochemicals were molecular docked against IL6, AKT1, EGFR, and CASP3 proteins. Safety was assessed by ADMET profiling, while cytotoxicity, migration, invasion, and gene expression effects of Kaempferol and Coumestrol were assessed by in vitro assays. Statistical analysis was conducted using SPSS software.Results:Kaempferol and Coumestrol were chosen from 69 phytochemicals based on high binding energies and atomic interactions with proteins. Coumestrol showed superior anti-invasive properties, and higher binding affinities with AKT1 (-6.4 kcal/mol), EGFR (-7.1 kcal/mol), and CASP3 (-7.1 kcal/mol) when compared to Kaempferol, plus no AMES toxicity and exceeded total clearance (8.085) of Kaempferol (6.868). Kaempferol exhibited greater cytotoxicity, colony-building inhibition, and stronger inhibition of cancer cell viability (38.8%) compared to Coumestrol (53.6%). Both bioactive compounds downregulated EGFR, AKT1, and IL6 while upregulating Caspase-3 expression, to efficiently target fibrogenic pathways.Conclusion:The study concluded that Euphorbia hirta(whole plant) and Lepidium sativum(seeds) have the potential for hepatoprotection for liver fibrosis.