Antioxidant and anti-inflammatory properties of Sparassis crispa extracts

Abstract

Aim: This study aimed to compare the antioxidant and anti-inflammatory properties ofSparassiscrispa (S.crispa) extracts prepared using different extraction methods and to evaluate how extraction conditions influence bioactive component profiles and biological activities. Methods: S.crispa was extracted using hot water (SC-HWE), high-temperature and high-pressure water (SC-HPWE), and 70% ethanol (SC-EE). Total polyphenol and flavonoid contents, β-glucan content, and antioxidant activities [2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP), superoxide dismutase (SOD)-like activity, and catalase-related activity] were evaluated. Anti-inflammatory effects were assessed in lipopolysaccharide (LPS)-stimulated RAW 264.7 (murine macrophage cell line) macrophages by measuring cell viability, nitric oxide (NO) production, and pro-inflammatory cytokine [interleukin (IL)-1β, IL-6, and tumor necrosis factor alpha (TNF-α)] production using enzyme-linked immunosorbent assay (ELISA). Cytokine levels were expressed as a percentage of the LPS-treated control. Results: Extraction methods significantly affected the composition and bioactivities ofS.crispa extracts. SC-EE exhibited the highest total polyphenol and flavonoid contents and showed higher DPPH radical scavenging activity and NO inhibitory effects. SC-HPWE contained the highest β-glucan content and demonstrated superior FRAP values along with notable NO inhibitory activity. All extracts reduced LPS-induced IL-1β, IL-6, and TNF-α production; IL-1β showed greater responsiveness to extract treatment, whereas TNF-α exhibited relatively modest changes. At higher concentrations, the suppressive effect on cytokine production was attenuated, indicating a modulatory rather than a strictly monotonic response. Under the present experimental conditions, quercetin showed a limited reduction in cytokine production. Conclusions: These results demonstrate thatS.crispa extracts exhibit extraction method-dependent antioxidant and anti-inflammatory activities. The observed effects may reflect the combined contributions of phenolic compounds and β-glucan rather than a single bioactive component.S.crispa extracts may serve as promising natural materials for functional applications related to oxidative stress and inflammation regulation.

Keywords

Sparassis crispa / extraction methods / antioxidant activity / anti-inflammatory activity / β-glucan / phenolic compounds / RAW 264.7 macrophages

References

[1] Rashidinejad A. The road ahead for functional foods: promising opportunities amidst industry challenges. Future Postharvest Food. 2024;1:266-73.

[2] Gulcin İ. Antioxidants: a comprehensive review. Arch Toxicol. 2025;99:1893-997.

[3] Kumar K, Mehra R, Guiné RPF, Lima MJ, Kumar N, Kaushik R, et al. Edible Mushrooms: A Comprehensive Review on Bioactive Compounds with Health Benefits and Processing Aspects. Foods. 2021;10:2996.

[4] You SW, Hoskin RT, Komarnytsky S, Moncada M. Mushrooms as functional and nutritious food ingredients for multiple applications. ACS Food Sci Technol. 2022;2:1184-95.

[5] Zhou Y, Chu M, Ahmadi F, Agar OT, Barrow CJ, Dunshea FR, et al. A comprehensive review on phytochemical profiling in mushrooms: occurrence, biological activities, applications and future prospective. Food Rev Int. 2024;40:924—51.

[6] Azmir J, Zaidul ISM, Rahman MM, Sharif KM, Mohamed A, Sahena F, et al. Techniques for extraction of bioactive compounds from plant materials: A review. J Food Eng. 2013;117:426—36.

[7] Dai J, Mumper RJ. Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules. 2010;15:7313—52.

[8] Cheung LM, Cheung PCK, Ooi VEC. Antioxidant activity and total phenolics of edible mushroom extracts. Food Chem. 2003;81:249-55.

[9] Ohno N, Miura NN, Nakajima M, Yadomae T. Antitumor 1,3—beta—glucan from cultured fruit body of Sparassis crispa. Biol Pharm Bull. 2000;23:866-72.

[10] Harada T, Miura N, Adachi Y, Nakajima M, Yadomae T, Ohno N. Effect of SCG, 1,3—β—D—Glucan fromSparassiscrispa on the Hematopoietic Response in Cyclophosphamide Induced Leukopenic Mice . Biol Pharm Bull. 2002;25:931—9.

[11] Oh DS, Kim HS, Shim BS, Wi AJ, Yoon BS, Kim KW, et al. Effect of mycelial culture of cauliflower mushroom (Sparassis crispa) using LED lighting. J Mushrooms. 2013;11:24-31.

[12] Kim MY, Seguin P, Ahn JK, Kim JJ, Chun SC, Kim EH, et al. Phenolic compound concentration and antioxidant activities of edible and medicinal mushrooms from Korea. J Agric Food Chem. 2008;56:7265—70.

[13] Choi MH, Han HK, Lee YJ, Jo HG, Shin HJ. In vitro anti—cancer activity of hydrophobic fractions of Sparassis latifolia extract using AGS, A529, and HepG2 cell lines. J Mushrooms. 2014;12:304-10. Korean.

[14] Folin O, Denis W. On phosphotungstic phosphomolybdic compounds as color reagents. J Biol Chem. 1912;12:239-43.

[15] Moreno MI, Isla MI, Sampietro AR, Vattuone MA. Comparison of the free radical—scavenging activity of propolis from several regions of Argentina. J Ethnopharmacol. 2000;71:109-14.

[16] Blois MS. Antioxidant determination by use of a stable free radical. Nature. 1958;181:1199-200.

[17] Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice—Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999;26:1231—7.

[18] Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem. 1996;239:70-6.

[19] Marklund S, Marklund G. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem. 1974;47:469-74.

[20] Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121-6.

[21] Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of a tetrazolium—based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 1987;47:936—42.

[22] Do QD, Angkawijaya AE, Tran—Nguyen PL, Huynh LH, Soetaredjo FE, Ismadji S, et al. Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica. J Food Drug Anal. 2014;22:296-302.

[23] Lee DS, Kim KH, Yook HS. Antioxidant activities of different parts of Sparassis crispa depending on extraction temperature. J Korean Soc Food Sci Nutr. 2016;45:1617—22. Korean.

[24] Shahidi F, Zhong Y. Measurement of antioxidant activity. J Funct Foods. 2015;18:757-81.

[25] Mishra KK, Pal RS, Arunkumar R, Chandrashekara C, Jain SK, Bhatt JC. Antioxidant properties of different edible mushroom species and increased bioconversion efficiency of Pleurotus eryngii using locally available casing materials. Food Chem. 2013;138:1557-63.

[26] Jeż M, Wiczkowski W, Zielińska D, Szawara—Nowak D, Białobrzewski I, Błaszczak W. The impact of high pressure processing on the phenolic profile, hydrophilic antioxidant and reducing capacity of purée obtained from commercial tomato varieties. Food Chem. 2018;261:201-9.

[27] Ferreira ICFR, Barros L, Abreu RMV. Antioxidants in wild mushrooms. Curr Med Chem. 2009;16:1543-60.

[28] Tanaka T, Narazaki M, Kishimoto T. IL—6 in inflammation, immunity, and disease. Cold Spring Harb Perspect Biol. 2014;6:a016295.

[29] Bradley JR. TNF—mediated inflammatory disease. J Pathol. 2008;214:149-60.

[30] Li Y, Yao J, Han C, Yang J, Chaudhry MT, Wang S, et al. Quercetin, Inflammation and Immunity. Nutrients. 2016;8:167.