Chemical profiling and in-vitro anti-inflammatory activity of bioactive fraction(s) from Trichodesma indicum (L.) R.Br. against LPS induced inflammation in RAW 264.7 murine macrophage cells.

Ethnopharmacological relevance
Trichodesma indicum (L.) R.Br. (family: Boraginaceae) is a medicinal herb largely used to
treat arthralgia, rheumatoid arthritis, wound healing, dysentery, etc. It’s mechanism of antiinflammatory
activity has not been systematically analyzed yet.
Aim of the study
The present study was undertaken to examine the anti-inflammatory effects of successive
solvent extracts (n-hexane extract (HE), ethyl acetate extract (EA), ethanol extract (EE),
aqueous extract (AE) and fractions of HE) from the aerial parts of Trichodesma indicum (TI)
against lipopolysaccharide (LPS) stimulated inflammatory reaction using mouse macrophage
RAW 264.7 cells.
Materials and methods
Cytotoxic effects of the extracts and fractions of TI were assessed by MTT assay. The effect
of extracts and fractions on the production of nitric oxide (NO) in RAW 264.7 macrophages
was measured using Griess reagent method. IL - 6, IL - 1β, TNF-α, iNOS and COX-2 gene
expressions were examined by a qRT-PCR method.
Results
RAW 264.7 macrophages pretreated with HE, EA, EE and AE of TI showed significant
decrease in the production of proinflammatory cytokines and NO without exhibiting
cytotoxicity. The potent HE was fractionated using flash chromatography into FA, FB, FC,
FD and FE. Among the five fractions, FE displayed a stronger ability to reduce IL - 1β, TNF-
α, iNOS, COX2 and NO importantly no cytotoxicity was observed. The phytochemical
compounds present in FE was further screened by Gas chromatography - Mass spectroscopy
(GC-MS). GC-MS analysis revealed that 1,2-benzenedicarboxylic acid diisooctyl ester as the
major compound in FE. Molecular docking analysis showed good inhibition of 1,2-
benzenedicarboxylic acid diisooctyl ester against TLR-4, NIK and TACE.
Conclusion
Our results suggested that 1,2-benzenedicarboxylic acid diisooctyl ester could be a potential
candidate in alleviating inflammatory reactions in TI.