Background: The current study was aimed to investigate the protective effect ofmethanolic extract ofJuniperussquamata leaf (JSME) against cyclophosphamide-induced immunosuppression in rats and the characterization of its bioactive compounds. JSME was first screened for its preliminary immunomodulatory activity throughin vitro lymphocyte proliferation assay and phagocytic activity of polymorphonuclear cells.
Methods: Immunoprotective of JSME was assessed through analysis of hematologicalparameters, phagocytic carbon clearance, and HA titer values in cyclophosphamide treated immunosuppressed Wistar albino rats. Silica gel column chromatography was used to isolate the bioactive fractions from JSME.
Results:Our in vitro studies revealed that JSME significantly enhanced lymphocyte proliferation as depicted from increased optical density values. It was also found to augment the phagocytic effect of the polymorphonuclear leucocytes by causing increased engulfment of Candida albicans cells. In the case of in vivo studies, cyclophosphamide was found to cause a noteworthy decline in the hemoglobin, RBCs, WBCs, and platelets count.Administration of JSME resulted in combating this myelosuppression by restoring hematological parameters to normal. Pretreatment with JSME also augmented the rate of clearance of carbon particles from the blood as depicted by a considerable reduction in the phagocytic index compared to cyclophosphamide treated group. JSME showed phagocytic index of 0.0497 ± 0.0063 and 0.0687±0.0029 at concentration of 100mg/kg and 200mg/kg body weight, respectively.
Conclusions: Three compounds (stigmasterol, apigenin, and luteolin) were isolated via chromatography techniques using different organic solvents.Further, the structures of the isolated compounds were elucidated by NMR. These compounds are being reported for the first time in Juniperus squamata. This work has to an extent, provided the pivot upon which future isolations and characterizations of the constituents of Juniperus squamata will anchor. Therefore, the constituents can serve as lead molecules for the development of agents with therapeutically useful immunogenic properties.