A Study Of Bi-directional Immunomodulatory Activity Of Trametes Versicolor Intra-and Exo-mycelial Polysaccharide Extracts On Raw264.7 Cells

This research studied the bi-directional immunomodulatory activity of Trametes versicolor intra- and exo- mycelial extracts- PSP, PSK and YZLY on murine macrophage cell line Raw264.7 in vitro. Their stimulation activities on NO production and phagocytosis on mouse Raw264.7 cell line were measured using Griess reaction and neural red method, and the inducible nitric oxide synthase (iNOS) levels were detected by Western Blotting method. Tumor necrosis factor-α(TNF-α)productions were measured by using Enzyme-Linked Immunosorbent Method (ELISA).The results showed that the phagocytic activities of the Raw264.7 cells were promoted after incubated with 5, 25, 100μg/mL of PSP, PSK, YZLY along for 24 hours respectively (p<0.01), and the highest phagocytic indexes were 1.29 (100μg/mL, PSP), 1.29 (5μg/mL, PSK), and 1.32 (5μg/mL, YZLY). These extracts also promoted NO release, and the extract concentration at 100μg/mL did the best, which the NO productions were significantly higher than that of control group (p<0.01). At the same time, the expression of iNOS was increased significantly. Compared to the control group, the TNF-αproduction was also promoted by PSP, PSK and YZLY significantly, while PSP, PSK and YZLY at the concentration 100μg/mL were the best. The productions of TNF-αat these respective extract concentrations were 24400, 30838, 26947 pg/mL respectively.LPS (1μg/mL) stimulated Raw 264.7 cells to produce a large amount of NO (23.11μM) after 24 hours. This action could be suppressed to some extent by the 5, 25, 100μg/mL of the PSP, YZLY (p<0.01). The maximum effective concentrations of PSP and YZLY were 25μg/mL, which could reduce the NO production to only 9.66 and 9.98μM respectively. PSK could also decreased the over NO production by LPS at concentrations of 25, 100μg/mL (20.86μM, 21.45μM, p<0.05). Similarly, these extracts could suppress over iNOS expression and TNF-αproduction stimulated by LPS. PSP, PSK and YZLY could reduce the LPS stimulated TNF-αproduction from 35923pg/mL to 29291 pg/mL (25μg/mL PSP), 31540 pg/mL (25μg/mL PSK) and 30697pg/mL (100μg/mL YZLY) respectively.In conclusion, the Trametes versicolor intra- and exo- mycelial extracts PSP, PSK and YZLY can regulate the mouse macrophage immune reactions in a bi-directional way. The up-regulation immunomodulatory activities in a strong to weak order is PSK, YZLY and PSP. The suppressing activity of these extracts on over immune reaction induced by LPS are however in a reverse order.