Water Soluble Antioxidative Crude Polysaccharide From Russula senecis Elicits TLR Modulated NF-κB Signaling Pathway and Pro-inflammatory Response in Murine Macrophages

Abstract

Russula senecis has recently been reported as a new addition to macrofungal flora of West Bengal. Besides, it also emerged as a seasonal health promoting nutrient to local ethnic people and enlisted for the first time as tribal food in our previous publication. In this context, the present work was designed to establish such usefulness scientifically and to meet the aim, crude polysaccharide, Rusenan, was prepared using conventional heated water reflux. Initially, the polymers were characterized to determine chemical composition and for that spectrophotometry along with Fourier-transform infrared spectroscopy (FT-IR), high-performance thin-layer chromatography (HPTLC), and gas chromatography-mass spectrometry (GC-MS) were performed. Analysis indicated that Rusenan was consisted mainly of carbohydrate conjugated with trace amount of protein. Furthermore, glucose was detected as the major monosaccharide (mainly in β-type glycosidic linkage) while other monomers were presented in the order of galactose > mannose > xylose > rhamnose. Conversely, antioxidant potential was determined following eight in vitro systems where the fraction evidenced strong superoxide, hydroxyl, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activity, high affinity to Fe2+ as well as instant ability to donate electron with EC50 values ranging from 80 to 3885 μg/ml concentration. In addition, effect on murine macrophages was also investigated where the polysaccharide treatment increased cell proliferation, phagocytic activity, filopodia or lamellipodia formation, nitric oxide (NO) production and reactive oxygen species (ROS) synthesis. Thereafter, through reverse transcriptase polymerase chain reaction (RT-PCR) analysis, significant increase in the expression of Toll like receptor (TLR)-4, TLR-2 and nuclear factor kappa B (NF-κB) was observed; as a result alleviated level of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, IκB-α, and interferon (IFN)-γ were also noticed explaining definite immune-stimulatory activity of the fraction. Thus, overall finding suggests that R. senecis can be considered as a functional food and may be used in preparation of dietary supplement to enhance general health.

Hot alkali‐extracted antioxidative crude polysaccharide from a novel mushroom enhances immune response via TLR‐mediated NF‐κB activation: A strategy for full utilization of a neglected tribal food

“…Thus, α‐linked glucan was perceived in trace as the extent was computed as 2.08 ± 0.17 g/100 g dry polysaccharide. Predominance of β‐glucan in carbohydrate backbone was in agreement with previous literatures (Khatua, & Acharya, 2016, 2018; Khatua, Dutta, et al., 2017); although the extents were found to be lesser than RualaHap.…”

“…Phenol sulfuric acid method was followed to detect polysaccharide distribution (Patra et al., 2013). Finally, helical structure of carbohydrate backbone was analyzed by characterizing Congo red‐polysaccharide reaction (Khatua & Acharya, 2018).…”

“…Effect of the polysaccharides on macrophage viability and phagocytic uptake was determined by using water soluble tetrazolium (WST) and neutral red reagents, respectively, after 24 as well as 48 hr incubation (Khatua, & Acharya, 2019b; Khatua, Dutta, et al., 2017). After 1 day treatment, nitric oxide (NO) production was estimated by Griess reagent (Khatua & Acharya, 2018). Real‐time NO production was also visualized using fluorochorme 4–5 diaminofluorescein diacetate (DAF‐2DA) dye (Zhou & He, 2011).…”

Evaluation of Polish Wild Mushrooms as Beta-Glucan Sources

“…The high β-glucan content of the genus Lactarius was also confirmed by Ruthes et al [ 72 ] and Hou et al [ 73 ]. Mushrooms of the genus Russula are rarely assessed for their β-glucan content; however, a few scientific reports confirm the high β-glucan content in their cell walls compared to other mushrooms [ 30 , 67 , 75 ].…”

Anti-Inflammatory and Antioxidant Effects of Carpesium cernuum L. Methanolic Extract in LPS-Stimulated RAW 264.7 Macrophages

“…HO-1 is an antioxidant enzyme induced by oxidative stress; however, overexpression of HO-1 prior to stimulation with LPS remarkably declined the production of subsequent inflammatory mediators such as NO, interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) [ 15 ]. In this respect, antioxidants are the first line of defense against oxidative damage induced by scavenging radicals and thus act to maintain optimal health [ 16 ].…”