Fig 1.
Assessing anti-inflammatory potential of I. obliquus in LPS-induced TNF-α production in RAW 264.7 macrophage cells.
E1 = 80% ethanol extract, E2 = 50% methanol extract, E3 = Water extract, E4 = 5% NaOH extract. Fungal conc. 0.25 μg/μL, PMB (100 units) was used as a positive control; DMEM and solvents were used as negative controls. Error bars are standard deviations. Value are means derived from triplicates (*, P < 0.05).
Fig 2.
Inhibition of histamine-induced TNF-α production.
In RAW 264.7 macrophage cells the mushroom extract I. obliquus (E2) was used. PMB (100 units) was used as a positive control and DMEM was used as negative control. The histamine concentration used is the dose for inducing maximum TNF-α- was 10−6 M. Error bars represent standard deviation (*, P < 0.05).
Table 1.
Baseline (resting) and maximum diameters (ACh and SNP) in 2A arteriole.
Fig 3.
Dose response curve of Acetylcholine (ACh) and Phenylephrine (PE) on second-order arterioles of gluteus maximus of male C57BL/6 mice (n = 10).
Values represent mean (± SEM) vessel diameter change calculated based on peak dilation or constriction as a percentage of resting diameter.
Fig 4.
Representative changes of arteriolar diameter following exposure of ACh or PE.
(A) baseline, (B) ACh 10-5M, (C) baseline, (D) PE 10-5M. Green arrow (arteriole), Blue arrow (Venule) and scale bar (μm).
Fig 5.
Dose response curve of histamine on second-order arterioles of gluteus maximus of male C57BL/6 mice (n = 5).
Values represent mean (± SEM) vessel diameter change calculated based on peak dilation or constriction as a percentage of resting diameter.
Fig 6.
Conducted vasodilation in arterioles of C57BL6 mice (n = 5).
(A)Responses to ACh (1psi, 500 ms) were recorded at the remote site upstream (with respect to blood flow) along the arteriole through a distance of 500 μm (conducted response). At each site, “diameter change” was calculated as peak response diameter–resting diameter. (B) I. obliquus (Chaga) reverses the reduction of the conducted response by histamine. There is significant difference between the control conducted response and histamine (a; p <0.05) and reversal of histamine vs. histamine + I. obliquus (Chaga) (b; P <0.07).
Fig 7.
Vasoactivity of compound 48/80 on second-order arterioles of gluteus maximus of male C57BL/6 mice.
C48/80 exhibits a biphasic dose-response pattern. Values represent mean (± SEM) vessel diameter change calculated based on peak dilation or constriction as a percentage of resting diameter (n = 5).
Fig 8.
Local and conducted vasodilation of second-order arterioles of male C57BL/6 mouse gluteus maximus.
In response to 1 M Ach in the presence of 10−6 M compound 48/80 the conducted vasodilation response is reduced and restored upon washout of the compound (n = 5). Values represent mean (± SEM) vessel diameter change calculated based on peak dilation or constriction as a percentage of resting diameter. *P < 0.05 vs. local dilation.
Fig 9.
Effect of histamine and compound 48/80 on the conduction of vasodilation in the presence of pyrilamine.
Pyrilamine abolished the effect of histamine on reducing conducted vasodilation but combination of C48/80 and pyrilamine, a significant reduction in conducted vasodilation was still observed. Values represent mean (± SEM) vessel diameter change calculated based on peak dilation or constriction as a percentage of resting diameter. Control value represents conducted dilation of vessels prior to treatment (n = 4 for each treatment). *P < 0.05 vs. control and histamine + pyrilamine treatment.