The polysaccharide isolated from <i>Pleurotus nebrodensis</i> (PN-S) shows immune-stimulating activity in RAW264.7 macrophages

CUI Hai-Yan; WANG Chang-Lu; WANG Yu-Rong; LI Zhen-Jing; ZHANG Ya-Nan

Volume 13 Issue 5

May 2015

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Article Navigation > Chinese Journal of Natural Medicines > 2015 > 13(5): 355-360

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The polysaccharide isolated from Pleurotus nebrodensis (PN-S) shows immune-stimulating activity in RAW264.7 macrophages

Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China

Corresponding author:
Received Date: 02-Apr.-2014

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 31101357 and 31271915).

Abstract

A novel Pleurotus nebrodensis polysaccharide (PN-S) was purified andcharacterized, and its immune-stimulating activity was evaluated in RAW264.7 macrophages. PN-S induced the proliferation of RAW264.7 cells in a dose-dependent manner, as determined by the MTT assay. After exposure to PN-S, the phagocytosis of the macrophages was significantly improved, with remarkable changes in morphology being observed. Flow cytometric analysis demonstrated that PN-S promoted RAW264.7 cells to progress through S and G2/M phases. PN-S treatment enhanced the productions of interleukin-6 (IL-6), nitric oxide (NO), interferon gamma (INF-), and tumor necrosis factor- (TNF-) in the macrophages, with up-regulation of mRNA expressions of interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), interferon gamma(INF-) and tumor necrosis factor- (TNF-) being observed in a dose-dependent manner, as measured by qRT-PCR. In conclusion, these results suggest that the purified PN-S can improve immunity by activating macrophages.
- Pleurotus nebrodensis,
- Polysaccharide,
- Immunization,
- Cytokines

References

[1]	Kim HG, Yoon DH, Kim CH, et al. Ethanol extracts of Inonotus obliquus inhibits lipopolysaccharide-induced inflammation in RAW 264.7 macrophage cells[J]. J Med Food, 2007, 10(1):80-89.
[2]	Song Y, Hui J, Kou W, et al. Identification of Inonotus obliquus and analysis of antioxidation and antitumor activities of polysaccharides[J]. Curr Microbiol, 2008, 57(5):454-462.
[3]	Qi C, Cai Y, Gunn L, et al. Differential pathways regulating innate and adaptive antitumor immune responses by particulate and soluble yeast-derived beta-glucans[J]. Blood, 2011, 117(25):6825-6836.
[4]	Ladanyi A, Timar J, Lapis K, et al. Effect of lentinan on macrophage cytotoxicity against metastatic tumor cells[J]. Cancer Immunol Immun, 1993, 36(2):123-126.
[5]	Chen X, Zhong HY, Zeng JH, et al. The pharmacological effect of polysaccharides from Lentinus edodes on the oxidative status and expression of VCAM-1mRNA of thoracic aorta endothelial cell in high-fat-diet rats[J]. Carbohyd Polym, 2008, 74(3):445-450.
[6]	Ai H, Wang FR, Yang Q, et al. Preparation and biological activities of chitosan from the larvae of housefly, Musca domestica[J]. Carbohyd Polym, 2008, 72(3):419-423.
[7]	Liu CH, Wang CH, Xu ZL, et al. Isolation, chemical characterization and antioxidant activities of two polysaccharides from the gel and the skin of Aloe barbadensis miller irrigated with sea water[J]. Process Biochem, 2007, 42(6):961-970.
[8]	Pang XB, Yao WB, Yang XB, et al. Purification, characterization and biological activity on hepatocytes of a polysaccharide from Flammulina velutipes mycelium[J]. Carbohyd Polym, 2007, 70(3):291-297.
[9]	Bao XF, Wang XS, Dong Q, et al. Structural features of immunologically active polysaccharides from Ganoderma lucidum[J]. Phytochemistry, 2002, 59(2):175-181.
[10]	Noriko M, Mitsuyo O, Shoji O, et al. Antihypertensive effect of Pleurotus nebrodensis in spontaneously hypertensive rat[J]. J Oleo Sci, 2008, 57(12):675-681.
[11]	Wang C, Cui H, Wang Y, et al. Bidirectional immunomodulatory activities of polysaccharides purified from Pleurotus nebrodensis[J]. Inflammation, 2014, 37(1):83-93.
[12]	Lee SM, Yoon MY, Park HR, et al. Protective effects of paeonia lactiflora pall on hydrogen peroxide-induced apoptosis in PC12 cells[J]. Biosci Biotechnol Biochem, 2008, 72(5):1272-1277.
[13]	Gordon S, The role of the macrophage in immune regulation[J]. Res Immunol, 1998,149(7-8):685-688.
[14]	Kim SA, Lim SS. T lymphocyte subpopulations and interleukin-2, interferon-c, and interleukin-4 in rat pulpitis experimentally induced by specific bacteria[J]. J Endodontics, 2002, 28(3):202-205.
[15]	Apetoh L, Ghiringhelli F, Tesniere A, et al. Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy[J]. Nat Med, 2007, 13(9):1050-1059.
[16]	Tamir S, Tannenbaum SR. The role of nitric oxide (NO-) in the carcinogenic process[J]. Biochim Biophys Acta (BBA)-Rev Cancer, 1996, 1288(2):F31-F36.
[17]	Lin H, Wei RQ, Bolling SF. Tumor necrosis factor- and interferon-cmodulation of nitric oxide and allograft survival[J]. J Surg Res, 1995, 59(1):103-110.
[18]	Yang AP, Zhang XL, Liu AP, et al. The effect of complex aloe polysaccharide films on oral ulceration of rat[J]. J Foshan Univ (Natural Science Edition), 2007, 25(3):46-48(in Chinese).
[19]	Djeraba A, Quere P. In vivo macrophage activation in chickens with acemannan, a complex carbohydrate extracted from Aloe vera[J]. Int J Immunopharmaco, 2000, 22(5):365-372.
[20]	Samodova AV, Dobrodeeva LK. Role of shedding in the activity of immunocompetent cells with reagin protection mechanism[J]. Fiziol Cheloveka, 2012, 38(4):114-120.
[21]	Bielak-Zmijewska A, Koronkiewicz M, Skierski J, et al. Effect of curcumin on the apoptosis of rodent and human nonproliferating and proliferating lymphoid cells[J]. Nutr Cancer, 2000, 38(1):131-138.

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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The polysaccharide isolated from Pleurotus nebrodensis (PN-S) shows immune-stimulating activity in RAW264.7 macrophages

Corresponding author:

1. Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China

Received Date: 2014-04-02

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 31101357 and 31271915).

Abstract: A novel Pleurotus nebrodensis polysaccharide (PN-S) was purified andcharacterized, and its immune-stimulating activity was evaluated in RAW264.7 macrophages. PN-S induced the proliferation of RAW264.7 cells in a dose-dependent manner, as determined by the MTT assay. After exposure to PN-S, the phagocytosis of the macrophages was significantly improved, with remarkable changes in morphology being observed. Flow cytometric analysis demonstrated that PN-S promoted RAW264.7 cells to progress through S and G2/M phases. PN-S treatment enhanced the productions of interleukin-6 (IL-6), nitric oxide (NO), interferon gamma (INF-), and tumor necrosis factor- (TNF-) in the macrophages, with up-regulation of mRNA expressions of interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), interferon gamma(INF-) and tumor necrosis factor- (TNF-) being observed in a dose-dependent manner, as measured by qRT-PCR. In conclusion, these results suggest that the purified PN-S can improve immunity by activating macrophages.

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Reference (21)

[1]	Kim HG, Yoon DH, Kim CH, et al. Ethanol extracts of Inonotus obliquus inhibits lipopolysaccharide-induced inflammation in RAW 264.7 macrophage cells[J]. J Med Food, 2007, 10(1):80-89.
[2]	Song Y, Hui J, Kou W, et al. Identification of Inonotus obliquus and analysis of antioxidation and antitumor activities of polysaccharides[J]. Curr Microbiol, 2008, 57(5):454-462.
[3]	Qi C, Cai Y, Gunn L, et al. Differential pathways regulating innate and adaptive antitumor immune responses by particulate and soluble yeast-derived beta-glucans[J]. Blood, 2011, 117(25):6825-6836.
[4]	Ladanyi A, Timar J, Lapis K, et al. Effect of lentinan on macrophage cytotoxicity against metastatic tumor cells[J]. Cancer Immunol Immun, 1993, 36(2):123-126.
[5]	Chen X, Zhong HY, Zeng JH, et al. The pharmacological effect of polysaccharides from Lentinus edodes on the oxidative status and expression of VCAM-1mRNA of thoracic aorta endothelial cell in high-fat-diet rats[J]. Carbohyd Polym, 2008, 74(3):445-450.
[6]	Ai H, Wang FR, Yang Q, et al. Preparation and biological activities of chitosan from the larvae of housefly, Musca domestica[J]. Carbohyd Polym, 2008, 72(3):419-423.
[7]	Liu CH, Wang CH, Xu ZL, et al. Isolation, chemical characterization and antioxidant activities of two polysaccharides from the gel and the skin of Aloe barbadensis miller irrigated with sea water[J]. Process Biochem, 2007, 42(6):961-970.
[8]	Pang XB, Yao WB, Yang XB, et al. Purification, characterization and biological activity on hepatocytes of a polysaccharide from Flammulina velutipes mycelium[J]. Carbohyd Polym, 2007, 70(3):291-297.
[9]	Bao XF, Wang XS, Dong Q, et al. Structural features of immunologically active polysaccharides from Ganoderma lucidum[J]. Phytochemistry, 2002, 59(2):175-181.
[10]	Noriko M, Mitsuyo O, Shoji O, et al. Antihypertensive effect of Pleurotus nebrodensis in spontaneously hypertensive rat[J]. J Oleo Sci, 2008, 57(12):675-681.
[11]	Wang C, Cui H, Wang Y, et al. Bidirectional immunomodulatory activities of polysaccharides purified from Pleurotus nebrodensis[J]. Inflammation, 2014, 37(1):83-93.
[12]	Lee SM, Yoon MY, Park HR, et al. Protective effects of paeonia lactiflora pall on hydrogen peroxide-induced apoptosis in PC12 cells[J]. Biosci Biotechnol Biochem, 2008, 72(5):1272-1277.
[13]	Gordon S, The role of the macrophage in immune regulation[J]. Res Immunol, 1998,149(7-8):685-688.
[14]	Kim SA, Lim SS. T lymphocyte subpopulations and interleukin-2, interferon-c, and interleukin-4 in rat pulpitis experimentally induced by specific bacteria[J]. J Endodontics, 2002, 28(3):202-205.
[15]	Apetoh L, Ghiringhelli F, Tesniere A, et al. Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy[J]. Nat Med, 2007, 13(9):1050-1059.
[16]	Tamir S, Tannenbaum SR. The role of nitric oxide (NO-) in the carcinogenic process[J]. Biochim Biophys Acta (BBA)-Rev Cancer, 1996, 1288(2):F31-F36.
[17]	Lin H, Wei RQ, Bolling SF. Tumor necrosis factor- and interferon-cmodulation of nitric oxide and allograft survival[J]. J Surg Res, 1995, 59(1):103-110.
[18]	Yang AP, Zhang XL, Liu AP, et al. The effect of complex aloe polysaccharide films on oral ulceration of rat[J]. J Foshan Univ (Natural Science Edition), 2007, 25(3):46-48(in Chinese).
[19]	Djeraba A, Quere P. In vivo macrophage activation in chickens with acemannan, a complex carbohydrate extracted from Aloe vera[J]. Int J Immunopharmaco, 2000, 22(5):365-372.
[20]	Samodova AV, Dobrodeeva LK. Role of shedding in the activity of immunocompetent cells with reagin protection mechanism[J]. Fiziol Cheloveka, 2012, 38(4):114-120.
[21]	Bielak-Zmijewska A, Koronkiewicz M, Skierski J, et al. Effect of curcumin on the apoptosis of rodent and human nonproliferating and proliferating lymphoid cells[J]. Nutr Cancer, 2000, 38(1):131-138.

The polysaccharide isolated from Pleurotus nebrodensis (PN-S) shows immune-stimulating activity in RAW264.7 macrophages

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The polysaccharide isolated from Pleurotus nebrodensis (PN-S) shows immune-stimulating activity in RAW264.7 macrophages

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1. Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China

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The polysaccharide isolated from Pleurotus nebrodensis (PN-S) shows immune-stimulating activity in RAW264.7 macrophages

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The polysaccharide isolated from Pleurotus nebrodensis (PN-S) shows immune-stimulating activity in RAW264.7 macrophages

Corresponding author:

1. Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China

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