Phytochemical screening and effects on cell-mediated immune response of Pleurotus fruiting bodies powder

Figures & data

Table 1. Phytochemical profile of Pleurotus sp. fruiting bodies powder.

Metabolites	Assays	Aqueous extract	Ethanolic extract
Alkaloids	Dragendorff	+++	+++
	Wagner	++	++
Terpenoids	Solkowski	−	+
	Lieberman–Burchard	−	+++
Reducing sugar	Fehling	+	+
	Benedict	+	+
Phenols and tannins	FeCl3	+	+
Amino acids	Ninhidrine	+	+++
Flavonoids	Concentrated H2SO4	+	+
	Rosemheim	+	+

Note: The phytochemicals contained in both aqueous and ethanolic extracts were estimated qualitatively according to Harbourne (1984). Three replicates were used for each assay.Legend: (−) none, (+) present, (++) mild, (+++) marked.

Table 2. Effect of therapeutic administration of Pleurotus powder on the delayed-type hypersensitivity response (foot pad thickness) of cyclophosphamide treated Balb/c mice.

	Foot pad thickness (mm)
	24 h	48 h	72 h
Control	0.48±0.07^a	0.46±0.04^a	0.38±0.07^a
CY-Pleurotus	0.14±0.05^b	0.43±0.02^a	0.29±0.05^a
CY-Saline	0.11±0.06^b	0.39±0.01^b	0.12±0.04^b

Note: Mice were immunised by an intradermal (i.d.) injection of 50 µl of 5 mg/ml bovine serum albumin (BSA) emulsified in Complete Freund Adjuvant (CFA) at two sites on the abdomen. Eight days after immunisation, the mice were rechallenged by injection of 20 µl of 5 mg/ml BSA into one rear foot pad, while the other received a comparable volume of PBS. Measurements of foot pad swelling were taken at 24, 48 and 72 h after challenge. The magnitude of the DTH response was determined as the differences in foot pad thickness between the antigen and PBS-injected foot pads. All values are expressed as the arithmetic mean±SD of five mice. Different letters indicate significant differences among groups (Kruskal-Wallis, Student–Newman–Keuls, p<0.05).

Figure 1. Mass index of popliteal lymph nodes of cyclophosphamide treated Balb/c mice administered or not with Pleurotus powder. The popliteal lymph nodes (right and left) of the antigen sensitised and rechallenged animals of DTH experiment were removed and weighed separately. The mass index was expressed as the relation between the weight of the popliteal node belonging to BSA-injected foot pad with respect to that of PBS-injected pad. The (*) reflects significant differences among the groups in the Mann–Whitney test (p<0.05).

Figure 2. In vitro lymphoproliferative-stimulating response of aqueous and ethanolic extracts obtained from Pleurotus powder on murine spleen cells. Phytohaemagglutinin (PHA; 200 µl) was added to tubes containing 5 ml of supplemented RPMI-1640 to a final concentration of 5 µg/tube, followed by the addition of 2×10⁶ cells and 100 µl of powder extracts (aqueous or ethanolic). The tubes were incubated at 37 °C for 72 h and then 500 µl of MTT (5 mg/ml) was added. After the incubation of the resulting mixture at 37 °C for 4 h, the tubes were centrifuged, the supernatants were discarded and 1 ml of isopropanol was added. The absorbance was measured at 570 nm. The experiments were done in triplicate. The stimulation index was calculated considering the absorbance of control cultures without PHA as the unit.