Identification of spathulenol in Salvia mirzayanii and the immunomodulatory effects

The methanol extract of Salvia mirzayanii has shown an immunomodulatory effect on peripheral blood lymphocytes. Bioassay-guided fractionation using a lymphocyte proliferation assay on Salvia mirzayanii was performed in order to purify and identify the active compounds. Fractionation of the methanol extract and purification of the components using normal column chromatography and preparative thin layer chromatography resulted in identification of the bioactive compound, spathulenol, with an immunoinhibitory effect. Identification of this compound was performed by 1D and 2D NMR methods and HRMS. Treatment of activated lymphocytes with a concentrated fraction containing 62% of spathulenol (SP) showed a decrease in the proliferation of lymphocytes with an IC(50) of 85.4 ± 11.08 µg/mL. Flow cytometry analysis using annexin V and propidium iodide staining of the stimulated peripheral blood lymphocytes in the presence of SP demonstrated a dose dependent increase in the percentage of apoptotic cells (IC(50) ; 77.2 ± 5.31 µg/mL). No significant increase in caspase 3 activity in a 20 h treatment of stimulated lymphocytes compared with the control was observed. In conclusion, this study identified the possible activity of spathulenol as one of the immunomodulatory compounds present in Salvia mirzayanii. SP showed the capacity to inhibit proliferation in the lymphocytes and to induce apoptosis in these cells possibly through a caspase-3 independent pathway.