Assessment of Antioxidant and Anti-Inflammatory Activities of T. erecta Leaf (Methanolic Extract ) Ointment in LPS-Stimulated RAW 264.7 Macrophages

ABSTRACT:Inflammation is natural response of the body to cell damage and stimuli. Inflammation andoxidative stress are key contributors to the cause and worsening of many chronic illnesses,such as arthritis, heart problems, and brain disorders.ManyThunbergiaspecies have long beenused in traditional medicine to treat a variety of conditions, such as fever andinflammation.Nevertheless, additional pharmacological validation is required to convert thisknowledge into standardized clinical practices, despite the potential of conventionalapplications and experimental studies.Present study was designed to carry out photochemicalscreening of methanolic extract of T. erecta(T. erecta) and to access its anti-inflammatoryactivity with the aid of in-vitro tests. Presence of secondary metabolites in test drug wasinvestigatedusing phytochemical screening tests.Antioxidant potential was evaluated usingFerric Reducing Antioxidant Power (FRAP) assay and nitric oxide scavenging assay (NO).By assessing the test drugs ability to prevent protein denaturation, anti-inflammatory activitywas determined preliminarily.To validate the anti-inflammatory potential at the cellular andmolecular level, RAW 264.7 macrophage cells were challenged with lipopolysaccharide(LPS) to establish an inflammatory microenvironment, followed by treatment with theT.erecta ointment (methanolic extract) test extract to evaluate its capacity to modulate theinflammatory response.Preliminary phytochemical screening confirmed the occurrence ofseveral bioactive secondary metabolites, including glycosides, steroids, flavonoids, alkaloids,phenols, proteins and carbohydrates alongside a notable capacity for free radical scavengingactivity.In two methods tested to evaluate the antioxidant activity, methanolic extractdisplayed the moderate antioxidant activity(IC50 value: 149.855 μ g/ml in nitrate scavenging assay).Less activity was seen with IC50value:287.88± 12.509 μ M/200μ gin FRAP assay). LPS-induced inflammatory mediators like COX-2 were inhibited by thetest drug in a dose-dependent manner.The test drug demonstrated anti-inflammatory propertyalong with antibacterial effect against Clostridium perfringens (gram-positive) bacteriathough its potency was lower than that of the standard drug.Nevertheless, the extractexhibited a notable anti-inflammatory response.