MOLECULAR AND CELLULAR NUTRITION

Effects of Ganoderma lucidum Polysaccharides on Immune Function of IPEC-1 Cells Infected with Escherichia coli

  • ZHAI Junlei ,
  • LIN Yin ,
  • WANG Meiyan ,
  • GAO Yuyun ,
  • WANG Changkang ,
  • JIN Ling
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  • 1. College of Animal Sciences(College of Bee Science), Fujian Agriculture and Forest University, Fuzhou 350002, China;
    2. China National Engineering Research Center of Juncao Technology, Fujian Agriculture and Forestry University, Fuzhou 350002, China

Received date: 2021-05-06

  Online published: 2021-12-16

Abstract

To explore the effects of Ganoderma lucidum polysaccharides on the proliferation, cycle and immune function of IPEC-1 cells, and to provide theoretical basis for the application of Ganoderma lucidum polysaccharides in intestinal health of piglets. The experiment was divided into blank group and different concentrations (5, 10, 20, 50, 100, 200, 400 and 800 μg/mL) of Ganoderma lucidum polysaccharides groups. The cell viability of IPEC-1 cells was detected by CCK-8 method, and the effects of Ganoderma lucidum polysaccharides on the cell cycle of IPEC-1 cells was detected by PI staining and flow cytometry. IPEC-1 cell inflammation model was constructed by enterohemorrhagic Escherichia coli (EHEC) and was used for subsequent experiments, which were divided into blank group, EHEC group, and different concentrations (5, 10, 20, 50 and 100 μg/mL) of Ganoderma lucidum polysaccharides+EHEC groups. The antimicrobial activity of Ganoderma lucidum polysaccharides treated IPEC-1 cells against EHEC was determined. The secretion amounts of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and interleukin-10 (IL-10) in the supernatant of IPEC-1 cells were determined by enzyme-linked immunosorbent assay (ELISA) method. The mRNA relative expression levels of IL-6, IL-1β, TNF-α and IL-10 in IPEC-1 cells were detected by fluorescence quantitative PCR. The results showed as follows:1) compared with the blank group, the cell viability decreased after 48 h of Ganoderma lucidum polysaccharides treatment, and the cell viability at 24 h was better than that at 12 h, so 24 h was selected as the best action time. Cell viability at 5, 10, 20, 50 and 100 μg/mL was better than 200, 400 and 800 μg/mL. 2) Ganoderma lucidum polysaccharides with the concentrations of 5, 10, 20, 50 and 100 μg/mL could significantly inhibit the proliferation of EHEC; IPEC-1 cells treated with 50 and 100 μg/mL Ganoderma lucidum polysaccharides for 24 h significantly improved the anti-EHEC activity (P<0.05). 3) Compared with the blank group, the secretion amounts of IL-1β, IL-6 and IFN-γ in EHEC group were significantly increased (P<0.05), while the secretion amount of IL-10 was significantly decreased; compared with the EHEC group, the secretion amounts of IL-6, TNF-α and IFN-γ in the 5, 10, 20, 50 and 100 μg/mL Ganoderma lucidum polysaccharides+EHEC groups was significantly decreased (P<0.05), the secretion amounts of IL-1β in the 20, 50 and 100 μg/mL Ganoderma lucidum polysaccharides+EHEC group were significantly decreased (P<0.05), and the secretion amount of IL-10 in 5 and 50 μg/mL Ganoderma lucidum polysaccharides+EHEC groups was significantly increased (P<0.05). 4) Compared with the blank group, the mRNA relative expression levels of IL-1β and TNF-α in EHEC group were significantly increased (P<0.05); compared with EHEC group, the mRNA relative expression levels of IL-1β in 50 and 100 μg/mL Ganoderma lucidum polysaccharides+EHEC groups was significantly decreased (P<0.05). It is concluded that Ganoderma lucidum polysaccharides can inhibit the proliferation of Escherichia coli, alleviate the inflammatory response of IPEC-1 cells and enhance the cellular immune function.

Cite this article

ZHAI Junlei , LIN Yin , WANG Meiyan , GAO Yuyun , WANG Changkang , JIN Ling . Effects of Ganoderma lucidum Polysaccharides on Immune Function of IPEC-1 Cells Infected with Escherichia coli[J]. Chinese Journal of Animal Nutrition, 2021 , 33(12) : 7118 -7130 . DOI: 10.3969/j.issn.1006-267x.2021.12.051

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