Downregulation of pro-inflammatory markers NF-κB1, RelA and COX-2 using Aconitum chasmanthum Stapf ex Holmes -in vitro and in-silico study

Abstract

Nuclear factor kappa B signaling pathway and cyclooxygenase-2 are considered molecular prototypes for targeting various diseases including inflammation. Aconitum chasmanthum has been used for decades to cure different ailments as its anti-inflammatory and anti-analgesic effects are well documented, but the mechanism remains unknown. Thus, in current study, we attempted to assess the anti-inflammatory characteristics of Aconitum chasmanthum which can be regarded as an appealing therapeutic candidate. In present study, High-Resolution Liquid Chromatography and Mass Spectrophotometry (HRLCMS-QTOF-MS) method were used to determine the phytoconstituents in the methanolic extract of wild rhizome of Aconitum chasmanthum. The in vitro studies were done on HCT-116 cell line and an MTT assay was used to assess the cell viability (IC50 value). In the present study, mRNA expression of pro-inflammatory genes (nuclear factor kappa B1, nuclear factor kappa B3 (RelA), and cyclooxygenase-2) was determined via quantitative polymerase chain reaction and western blot. Molecular docking analysis was performed by Auto Dock Vina to find potential compounds having anti-cyclooxygenase-2 activity. The current investigations found that methanolic extract strongly downregulates the gene expression of pro-inflammatory genes at a concentration of 200 μg/mL. While as western blot results showed a significant decrease in cyclooxygenase-2 protein level at 200 μg/mL concentration for 48 hrs. ADMET analysis revealed the physiochemical properties of bioactives and a drug. Molecular docking revealed staurosporine was the most effective compound against COX-2 with the highest binding affinity of − 9.7 kcal/mol. Thus the findings of current study reveal that Aconitum chasmanthum possesses substantial anti-inflammatory/anti-analgesic properties and therefore can be used to develop novel anti-inflammatory and anti-analgesic drugs in future.