422 Immunostimulant activity of Safed Musli Rate of Carbon Clearance Rate of carbon clearance is the measure of competency of the reticuloendothelial system and its granulopoetic activity (14), the faster removal of carbon particles has been correlated with the enhanced phagocytic activity. In the present study an increased phagocytic activity was observed in treated groups as compared to control. The rate of carbon clearance which was determined as phagocytic index was 20 % (1.2 ± 0.03) and 25% (1.36 ± 0.51) greater in sapogenins and ethanolic extract-treated groups, respectively, clearly indicating an enhanced in vivo phagocytic activity (Table 1). Table 1. Effect of administration of C. borivilianum ethanolic extract/ sapogenins on non-specific and specific immune systems Groups Control Test Group I Test Group II Survival rate percentage 33.3 ± 9.642 94.43 ± 5.56b 88.86 ± 5.56b DTH response (% increase in foot pad thickness) 0.89 ± 0.04 1.51 ± 0.02b (69.6%) 1.34 ± 0.07a (50.6%) Phagocytic Index (Ksample/ Kstandard) 1 1.36 ± 0.51b 1.2 ± 0.03b aP < 0.05 considered significant. bP < 0.005 considered extremely significant. Control: no treatment. Test Group II: C. borivilianum ethanol extract treated group (200 mg/kg b.w. p.o.). Test Group III: C. borivilianum sapogenin treated group (100 mg/kg b.w. p.o.). Table 2. Effect of administration of C. borivilianum ethanolic extract/ sapogenins on non-specific and specific immune systems Groups Test Group I Test Group II Haemoglobin concentration (g%) 14.5 ± 0.31 15.3 ± 0.25b Platelet count in thousand per cmm 468.5 ± 0.51 578.5 ± 0.56c RBC count in millions per cmm WBC count in thousand per cmm 4.7 ± 0.31 10.1 ± 0.38 5.9 ± 0.57a 11.3 ± 0.47a Test Group III 14.7 ± 0.48a Test Group IV 11.9 ± 0.32c 496.6 ± 0.56 5.2 ± 0.12 10.8 ± 0.28a 300.6 ± 0.24c 3.13 ± 0.43b 9.33 ± 0.45a Test Group V 14.3 ± 0.11a 560.45 ± 0.42c 5.99 ± 0.35b 10.8 ± 0.38a Test Group VI 14.0 ± 0.09a 524.65 ± 0.39b 5.86 ± 0.31b 10.5 ± 0.46a Test Group I: control (no treatment); Test Group II: ethanolic extract 200 mg per kg b.w; Test Group III: sapogenin extract 100 mg per kg b.w; Test Group IV: azathioprine 100 mg per kg b.w; Test Group V: azathioprine 100 mg per kg b.w; and ethanolic extract 200 mg per kg b.w; Test Group VI: azathioprine 100 mg per kg b.w. and Sapogenin extract 100 mg per kg b.w. aP > 0.5 not significant. bP < 0.05 significant. cP < 0.005 extremely significant. Effectiveness Against Drug-Induced Immunosuppression Administration of ethanolic extracts and sapogenins in general had a beneficial effect on hematological profile wherein all the parameters such as hemoglobin, platelets, RBC and WBC counts were increased. Simultaneous administration of etha- nolic extract and sapogenins along with azathioprine resulted in restoration of suppressed values observed after azathioprine treatment alone. Platelet, hemoglobin, RBC and WBC values observed were better than untreated control groups. Among the two test drugs ethanolic extract was pronouncedly more effective (P < 0.005) followed by sapogenin treatment (P < 0.05) (Fig. 1 and Table 2). Neutrophil Adhesion Test This test is an indicative of the marginalization of phagocytic cells in the blood vessels, i.e. an indication of immunostimula- tion. The % neutrophil adhesion in control group animals was 23.34 ± 2.1, in ethanolic extract-treated group it was 27.21 ± 1.6 whilst for sapogenin extract-treated group it was 25.62 ± 1.4. As is evident from the results of neutrophil adhesion test, nearly 10% (P < 0.5) increase in neutrophil adhesion is observed after administration of sapogenin extract, whilst a significant 17% (P < 0.05) increase in neutrophil adhesion is observed in ethanolic extract (Table 3). Discussion The scientific evidences collected from the study supports the traditional claims behind usage of the herb C. borivilianum, which is being cultivated and marketed extensively in India and abroad for medicinal purposes (12). The study affirms that C. borivilianum root extract is an effective immunostimulatory principle. The inference that can be drawn from the present study is that the total ethanolic extract is superior over sapogenin fraction of the plant as far as immunostimulatory activity is concerned. The extract does not only potentiate non- specific immune response, but is also effective in improving humoral as well as cell-mediated immunity. The effectiveness of extract-treated animals in overcoming the side effects of drug-induced immunosuppression provides evidence for bal- ancing and adaptogenic effectiveness of extracts. Table 3. Effect of C. borivilianum ethanolic extract/sapogenins on neutrophil adhesion in rats (mean ± SE) Animal group Group I Group II Group III TLC (103 mm 3) (X) % Neutrophil (Y) Neutrophil Index (X Y) Un B 6.4 ± 2.8 6.9 ± 1.4 6.8 ± 1.2 FTB 5.9 ± 2.2 6.1 ± 1.6 6.1 ± 1.4 Un B 44 ± 2.23 52 ± 2.21 51 ± 2.23 FTB 37 ± 3.2 43 ± 1.38 44 ± 1.47 Un B 284.8 ± 35.1 362.2 ± 32.4 356.6 ± 31.4 FTB 218.3 ± 25.4 265.1 ± 31.2 268.8 ± 32.1 % Neutrophil adhesion 23.34 ± 2.1 27.21 ± 1.6b 25.62 ± 1.4a UnB: untreated blood; FTB: nylon fiber treated blood; Group I: control (no treatment); Group II: C. borivilianum ethanolic extract 200 mg per kg b.w; Group III: C. borivilianum sapogenin extract 200 mg per kg b.w. aP < 0. 5 not significant. bP < 0.05 considered significant.