Saccharum Alhagi polysaccharide-1 and -2 promote the immunocompetence of RAW264.7 macrophages in vitro

Li,Gairu; Xiang,Yang; Zhao,Jin; Chang,Junmin

doi:10.3892/etm.2018.5818

Saccharum Alhagi polysaccharide-1 and -2 promote the immunocompetence of RAW264.7 macrophages in vitro

Authors:
- Gairu Li
- Yang Xiang
- Jin Zhao
- Junmin Chang
View Affiliations

Affiliations: Department of Pharmacy, Xinjiang Medical University, Urumqi, Xinjiang 830011, P.R. China, Center for Medical Laboratory, Clinical Medical Research Institute, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830054, P.R. China, Department of Preclinical Medicine, Shihezi Health School, Shihezi, Xinjiang 832000, P.R. China
Published online on: January 31, 2018 https://doi.org/10.3892/etm.2018.5818
Pages: 3556-3562

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The in vitro immune activities of Saccharum Alhagi polysaccharides (SAP) have been previously studied. The present study aimed to investigate the effects of SAP‑1 and SAP‑2 on the activity of RAW264.7 mouse macrophages. RAW264.7 cells were treated with 150, 300 and 600 mg/l concentrations of SAP‑1 (a 50% alcohol precipitation) and SAP‑2 (an 80% alcohol precipitation) or with 10 mg/l lipopolysaccharide. Untreated cells were used as a negative control. An MTT assay was used to detect the proliferation of the cells, and Hoechst 33528 staining was conducted in order to visualize the cell nuclei. Additionally, the Griess method was used to measure nitric oxide (NO) levels. A neutral red uptake assay was performed to determine the phagocytic activity of the macrophages, and ELISAs were performed to detect cytokine levels. Furthermore, reverse transcription‑quantitative polymerase chain reaction was used to measure the mRNA expression of certain cytokines. The results demonstrated that SAP increased the proliferative activity and activated the immune function of RAW264.7 cells, and was lacking in cytotoxicity. In addition, SAP‑1 exhibited a stronger effect in promoting RAW264.7 cell proliferation than did SAP‑2. Furthermore, SAP‑1 and SAP‑2 significantly increased the level of NO, with the effect of SAP‑1 being stronger than that of SAP‑2. SAP‑1 increased the phagocytic activity of RAW264.7 cells and promoted the secretion of the cytokines interleukin (IL)‑1β, IL‑2 and tumor necrosis factor (TNF)‑α by RAW264.7 cells, with an effect that was stronger than that of SAP‑2. Finally, different concentrations of SAP‑1 or SAP‑2 had distinct effects in upregulating the expression of TNF‑α, IL‑1β, nuclear factor‑κB and inducible nitric oxide synthase mRNA. The results of the present study demonstrate that SAP is capable of enhancing the immune activity of mouse macrophages.

View Figures

View References

1	Rukeyamu Shadike: Commonly used herbs in Uygur medicine (Written in Uygur). Xinjiang Science and Technology Press; Urumqi: pp. 2841993
2	Wu J, Li GR and Chang JM: Study on the extraction technology of polysaccharides from Saccharum Alhagi. Chin Traditional Patent Med. 33:9032011.(In Chinese).
3	Gong Y, Wu J and Li ST: Immuno-enhancement effects of Lycium ruthenicum Murr. Polysaccharide on cyclophamide-induced immunosuppression in mice. Int J Clin Exp Med. 8:20631–20637. 2015.PubMed/NCBI
4	Sun H, Zhang J, Chen F, Chen X, Zhou Z and Wang H: Activation of RAW264.7 macrophages by the polysaccharide from the roots of Actinidia eriantha and its molecular mechanisms. Carbohydr Polym. 121:388–402. 2015. View Article : Google Scholar : PubMed/NCBI
5	Brynjolfsson SF, Henneken M, Bjarnarson SP, Mori E, Del Giudice G and Jonsdottir I: Hyporesponsiveness following booster immunization with bacterial polysaccharides is caused by apoptosis of memory B cells. J Infect Dis. 205:422–430. 2012. View Article : Google Scholar : PubMed/NCBI
6	Yao L, Wang Z, Zhao H, Cheng C, Fu X, Liu J and Yang X: Protective effects of polysaccharides from soybean meal against X-ray radiation induced damage in mouse spllen lymphocytes. Int J Mol Sci. 12:8096–8104. 2011. View Article : Google Scholar : PubMed/NCBI
7	Jin Y, Tachibana I, Takeda Y, He P, Kang S, Suzuki M, Kuhara H, Tetsumoto S, Tsujino K, Minami T, et al: Statins decrease lung inflammation in mice by upregulating tetraspanin CD9 in macrophages. PLoS One. 8:e737062013. View Article : Google Scholar : PubMed/NCBI
8	Wu M, Luo X, Xu X, Wei W, Yu M, Jiang N, Ye L, Yang Z and Fei X: Antioxidant and immunomodulatory activities of a polysaccharide from Flammulina velutipes. J Tradit Chin Med. 34:733–740. 2014. View Article : Google Scholar : PubMed/NCBI
9	Zhao J, Li GR and Zheng J: Screening of polysaccharides with antioxidant activities from Saccharum Alhagi. J Xinjiang Med Univ. 38:1479–1481. 2015.(In Chinese).
10	Han Z, He J and Chang J: The influence of polysaccharides from Saccharum Alhagi on the immune activity of macrophage cell line RAW264.7. J Xinjiang Med Univ. 40:361–365. 2017.(In Chinese).
11	Jian L, Li G and Chang J: Determination of monosaccharide composition in polysaccharide of alhagi-honey by pre-column derivatization-high performance capillary electrophoresis. Chin J N Drugs. 21:78–81. 2012.
12	Su KY, Yu CY, Chen YP, Hua KF and Chen YL: 3,4-dihydroxytoluene, a metabolite of rutin, inhibits inflammatory responses in lipopolysaccharide-activated macrophages by reducing the activation of NF-κB signaling. BMC Complement Altern Med. 14:212014. View Article : Google Scholar : PubMed/NCBI
13	Kim KS, Cui X, Lee DS, Sohn JH, Yim JH, Kim YC and Oh H: Anti-inflammatory effect of neoechinulin a from the marine fungus Eurotium sp.SF-5989 through the Suppression of NF-кB and p38 MAPK pathways in lipopolysaccharide stimulated RAW264.7 macrophages. Molecules. 18:13245–13259. 2013. View Article : Google Scholar : PubMed/NCBI
14	Etoh T, Kim YP, Ohsaki A, Komiyama K and Hayashi M: Inhibitory effect of erythraline on Toll-like receptor signaling pathway in RAW264.7 cells. Biol Pharm Bull. 36:1363–1369. 2013. View Article : Google Scholar : PubMed/NCBI
15	Gao Y, Liu F, Fang L, Cai R, Zong C and Qi Y: Genkwanin inhibits proinflammatory mediators mainly through the regulation of miR-101/MKP-1/MAPK pathway in LPS-activated macrophages. PLoS One. 9:e967412014. View Article : Google Scholar : PubMed/NCBI
16	Zhao Q, Qian Y, Li R, Tan B, Han H, Liu M, Qian M and Du B: Norcantharidin facilitates LPS-mediated immune responses by up-regulation of AKT/NF-kB signaling in macrophages. PLoS One. 7:e449562012. View Article : Google Scholar : PubMed/NCBI
17	Chen JL, Zhong WJ, Tang GH, Li J, Zhao ZM, Yang DP and Jiang L: Norditerpenoids from Flickingeria fimbriata and their inhibitory activities on nitric oxide and tumor necrosis factor-α production in mouse macrophages. Molecules. 19:5863–5875. 2014. View Article : Google Scholar : PubMed/NCBI
18	O'Mahony C, Scully P, O'Mahony D, Murphy S, O'Brien F, Lyons A, Sherlock G, MacSharry J, Kiely B, Shanahan F and O'Mahony L: Commensal-induced regulatory T cells mediate protection against pathogen-stimulated NF-kappaB activation. PLoS Pathog. 4:e10001122008. View Article : Google Scholar : PubMed/NCBI
19	Nishitani Y, Zhang L, Yoshida M, Azuma T, Kanazawa K, Hashimoto T and Mizuno M: Intestinal anti-inflammatory activity of lentinan: Influence on IL-8 and TNFR1 expression in intestinal epithelial cells. PLoS One. 8:e624412013. View Article : Google Scholar : PubMed/NCBI
20	Poormasjedi-Meibod MS, Jalili RB, Hosseini-Tabatabaei A, Hartwell R and Ghahary A: Immuno-regulatory function of indoleamine 2,3 dioxygenase through modulation of innate immune responses. PLoS One. 8:e710442013. View Article : Google Scholar : PubMed/NCBI
21	Ohnishi T, Bandow K, Kakimoto K, Kusuyama J and Matsuguchi T: Long-Time treatment by low-dose N-Acetyl-L-Cysteine enhances proinflammatory cytokine expressions in LPS-stimulated macrophages. PLoS One. 9:e872292014. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
23	Liang Z, Zeng Y, Huang X and Yang Z: The effect of Apigenin on proliferation and No secretion and phagocytosis of RAW264.7 cells. J Jinan Univ (Natural Sci). 29:95–98. 2008.(In Chinese).
24	Xie Y, Chen Q, Luo D and Zhong Z: Anti-fatigue and immunoregulatory functions of high-purity rubusoside. Lishizhen Med Materia Med Res. 21:1421–1422. 2010.(In Chinese).
25	Huang F, Guo Y, Zhang R, Yi Y, Deng Y, Su D and Zhang M: Effects of drying methods on physicochemical and immunomodulatory properties of polysaccharide-protein complexes from litchi pulp. Molecules. 19:12760–12776. 2014. View Article : Google Scholar : PubMed/NCBI
26	Luo J, Zhu R, Yi L Dong Y and Wang PX: Effect of sinomenine on mouse RAW264.7 macrophage cells line polarization induced by LPS or IL-4. Chin J Immunol. 31:56–60. 2015.
27	Byeon SE, Lee J, Kim JH, Yang WS, Kwak YS, Kim SY, Choung ES, Rhee MH and Cho JY: Molecular mechanism of macrophage activation by red ginseng acidic polysaccharide from Korean red ginseng. Mediators Inflamm. 2012:1–7. 2012. View Article : Google Scholar