In Vitro Immune Response of Saponin Rich Fraction of Bacopa Monnieri, Linn

The saponin rich fraction (SRF) of Bacopa monnieri were prepared and evaluated for in vitro immune response using murine immune cells. Thioglycollate medium elicited peritoneal exudates cells (PEC) were used to study in vitro effects of the fraction (SRF) on release of immune mediators. SRF was evaluated at different concentrations (832 – 6.5μg/ml) for release of nitric oxide (NO), superoxide (NBT reduction), lysosomal and myeloperoxidase assays on PEC. SRF was also screened for cytotoxicity in sulforhodamine B (SRB) assay on murine isolated peritoneal macrophages, splenocytes and bone marrow cells. The data was statistically compared by using one way ANOVA followed by Dunnet’s multiple comparisons test. The fraction showed significant (P<0.05) stimulation of release of NO at 416μg/ml (stimulation index, SI=1.67) and 208μg/ml (SI=2.14). NBT reduction were significantly stimulated at 52μg/ml (SI=1.34) by the fraction as compared to control (SI=1). Lysosomal enzyme activity was stimulated by the fraction at 104μg/ml (SI=1.89). Myeloperoxidase activity were significantly increased at 832μg/ml (SI=1.43) and 416μg/ml (SI=1.36) of the fraction with respect to control wells (SI=1). Phytohemagglutinin-M (positive control) showed stimulation of release of all the tested mediators. The SRF showed stimulation of macrophages (208μg/ml), splenocytes (832 and 416μg/ml) and bone marrow cells (208μg/ml) in SRB assay. In conclusion, saponin rich fraction of Bacopa exhibited in vitro stimulatory effects on release of immune mediators from murine peritoneal cells and proliferative effects on immune cells in vitro.