Steroidal saponins of Trillium govanianum: Quality control, pharmacokinetic analysis, and anti-inflammatory activity

Abstract

The rhizomes of Trillium govanianum have traditionally been used in the Ayurvedic system of medicine to treat inflammation, pain, burn, and reproductive malfunctions. However, the ethanopharmacology and chemical constituents of T. govanianum have not been effusively explored to date. The present study attempted to uncover the phytochemical constituents and quality parameters of T. govanianum rhizomes, along with an evaluation of the anti-inflammatory potential of the extract, fractions, and isolated steroidal saponins. The quality of T. govanianum rhizomes was examined by standard guidelines of the Association of Official Agricultural Chemists (AOAC). The phytochemical composition of rhizomes was estimated by the spectroscopic method. The extract, fractions, and pure molecules were tested for their anti-inflammatory potential using lipopolysaccharide-stimulated RAW 264.7 macrophages. Besides that, computational pharmacokinetics properties of bioactive compounds were screened by using the Schrödinger software. The quality control parameters such as moisture content, ash value, pH, heavy metals, alpha toxins, residual pesticides, and microbial load were observed within the permitted limits as per AOAC guidelines, suggesting good quality and safe nature of T. govanianum rhizomes. Phytochemical analysis revealed the presence of steroidal saponins (78.3 ± 0.61 mg/g), phenolics (12.3 ± 2.76 mg/g), flavonoids (3.54 ± 0.24 mg/g), and carbohydrates (473 ± 2.27 mg/g) in the rhizomes. Macrophages cytocompatibility demonstrated that cell viability was not affected at any tested concentration of extract, fractions and pure compounds [diosgenin ( 1 ), borassoside D ( 2 ), and govanoside B ( 7 )]. The LPS-stimulated macrophages secreted nitric oxide (NO) and pro-inflammatory cytokines (TNF α , IL-1 β, and IL-6), which were inhibited by the extract, fractions, and compounds 1 , 2 , and 7 in a concentration-independent manner. Additionally, in silico pharmacokinetics revealed that compound 1 has good membrane-permeability followed by compound 2 , while compound 7 showed the non-membrane permeability. This study validates the traditional uses of rhizomes of T. govanianum in the treatment of inflammation, and this activity might be due to the presence of steroidal saponins . • Assessment of different quality control parameters of Trillium govanianum and all were observed within the permitted limits as per AOAC guidelines. • Phytochemical analysis revealed saponins (78.3 ± 0.61 mg/g), as the major constituents of this plant. • Anti-inflammatory and in silico pharmacokinetics activity evaluation of the extract, fractions, and three pure steroidal saponins of T. govanianum using lipopolysaccharide-stimulated RAW 264.7 macrophages. • Pure molecules effectively inhibit the production of cytokines (TNF- α , IL1 β , and IL-6) as compared with extract and fractions.