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Anti-fatigue effect of fermented porcine placenta through the regulation of fatigue-associated inflammatory cytokines
  • 비영리 CC BY-NC
  • 비영리 CC BY-NC
ABSTRACT

Fatigue is a common complaint and affects the quality of life in modern people. Physical stress may induce activation of certain immune cells. Fermented porcine placenta (FPP) has been used to alleviate fatigue. Inflammatory cytokines are produced by physical stress and results in symptoms of fatigue. However, the role of FPP on fatigue-associated inflammatory cytokine production has not been elucidated yet. Thus, we estimated the anti-fatigue effect of FPP and its active components, leucine (Leu) and lysine (Lys) in activated RAW264.7 macrophages and forced swimming test (FST) fatigue animal model. Pretreatment with FPP, Leu, or Lys significantly inhibited the lipopolysaccharide (LPS)-induced tumor necrosis factor-α, interleukin (IL)-1β, and IL-6 production without inducing cytotoxicity on LPS-stimulated RAW264.7 macrophages. FPP, Leu, or Lys inhibited the production of nitric oxide and downregulated the expression of inducible nitric oxide synthase on LPS-stimulated RAW264.7 macrophages. Furthermore, caspase-1 activities increased by LPS were significantly reduced by FPP, Leu, or Lys. In the FST, inflammatory cytokine levels of the mice administrated with FPP, Lys, and Leu were significantly reduced compared with the control group at 21 days. Collectively, these results show that anti-fatigue effect of FPP and its active components, Leu and Lys might be derived from the down-regulating of inflammatory mediators.


KEYWORD
fermented porcine placenta , inflammatory cytokine , caspase-1 , nitric oxide , forced swimming test
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이미지 / 테이블
  • [ Fig. 1. ]  FPP inhibits the productions of TNF-α, IL-1β, and IL-6 in RAW264.7 macrophages. Cells were pretreated with FPP, Leu, or Lys for 1 h and then stimulated with LPS (10 μg/ml) for 24 h. (A-C) The levels of TNF-α, IL-1β, and IL-6 were measured by ELISA method. (D) Cell viability was determined with an MTT assay. Data are mean ± SEM values of three independent experiments performed in duplicate. #p< 0.05: significantly different from the unstimulated cells; *p< 0.05: significantly different from the LPS-stimulated cells. LPS, lipopolysaccharide; FPP, fermented porcine placenta; Leu, leucine; Lys, lysine.
    FPP inhibits the productions of TNF-α, IL-1β, and IL-6 in RAW264.7 macrophages. Cells were pretreated with FPP, Leu, or Lys for 1 h and then stimulated with LPS (10 μg/ml) for 24 h. (A-C) The levels of TNF-α, IL-1β, and IL-6 were measured by ELISA method. (D) Cell viability was determined with an MTT assay. Data are mean ± SEM values of three independent experiments performed in duplicate. #p< 0.05: significantly different from the unstimulated cells; *p< 0.05: significantly different from the LPS-stimulated cells. LPS, lipopolysaccharide; FPP, fermented porcine placenta; Leu, leucine; Lys, lysine.
  • [ Fig. 2. ]  FPP inhibits the NO production and iNOS expression in RAW264.7 macrophages. Cells were pretreated with FPP, Leu, or Lys for 1 h and then stimulated with LPS (10 μg/ml) for 48 h. (A) NO production was measured by Griess method. Data are mean ± SEM values of three independent experiments performed in duplicate. Cells were pretreated with FPP, Leu, or Lys for 1 h and then stimulated with LPS (10 μg/ml) for 24 h. The total proteins were determined for iNOS expression by Western blotting. Results are representative of three independent experiments. #p< 0.05: significantly different from the unstimulated cells; *p< 0.05: significantly different from the LPS-stimulated cells. LPS, lipopolysaccharide; FPP, fermented porcine placenta; Leu, leucine; Lys, lysine.
    FPP inhibits the NO production and iNOS expression in RAW264.7 macrophages. Cells were pretreated with FPP, Leu, or Lys for 1 h and then stimulated with LPS (10 μg/ml) for 48 h. (A) NO production was measured by Griess method. Data are mean ± SEM values of three independent experiments performed in duplicate. Cells were pretreated with FPP, Leu, or Lys for 1 h and then stimulated with LPS (10 μg/ml) for 24 h. The total proteins were determined for iNOS expression by Western blotting. Results are representative of three independent experiments. #p< 0.05: significantly different from the unstimulated cells; *p< 0.05: significantly different from the LPS-stimulated cells. LPS, lipopolysaccharide; FPP, fermented porcine placenta; Leu, leucine; Lys, lysine.
  • [ Fig. 3. ]  FPP inhibits the caspase-1 activity in RAW264.7 macrophages. Cells were pretreated with FPP, Leu, or Lys for 1 h and then stimulated with LPS (10 μg/ml) for 2 h. (A) Caspase-1 activity was measured by a caspase-1 assay kit. Data are mean ± SEM values of three independent experiments performed in duplicate. (B) Caspase-1 protein levels were analyzed by Western blotting. Results are representative of three independent experiments. #p< 0.05: significantly different from the unstimulated cells; *p< 0.05: significantly different from the LPS-stimulated cells. LPS, lipopolysaccharide; FPP, fermented porcine placenta; Leu, leucine; Lys, lysine.
    FPP inhibits the caspase-1 activity in RAW264.7 macrophages. Cells were pretreated with FPP, Leu, or Lys for 1 h and then stimulated with LPS (10 μg/ml) for 2 h. (A) Caspase-1 activity was measured by a caspase-1 assay kit. Data are mean ± SEM values of three independent experiments performed in duplicate. (B) Caspase-1 protein levels were analyzed by Western blotting. Results are representative of three independent experiments. #p< 0.05: significantly different from the unstimulated cells; *p< 0.05: significantly different from the LPS-stimulated cells. LPS, lipopolysaccharide; FPP, fermented porcine placenta; Leu, leucine; Lys, lysine.
  • [ Fig. 4. ]  FPP inhibits the fatigue-related inflammatory cytokines in serum and spleen after FST. After the FST, (A-C) the levels of TNF-α, IL-1β, and IL-6 in the serum were analyzed by ELISA method. (D-F) The levels of TNF-α, IL-1β,and IL-6 in the spleen were analyzed by ELISA method. Cytokine levels in the spleen were divided according to the total protein levels. Values are the mean ± SEM. FPP: fermented porcine placenta, Lys: lysine, Leu: leucine. *p< 0.05, significantly different from the control mice.
    FPP inhibits the fatigue-related inflammatory cytokines in serum and spleen after FST. After the FST, (A-C) the levels of TNF-α, IL-1β, and IL-6 in the serum were analyzed by ELISA method. (D-F) The levels of TNF-α, IL-1β,and IL-6 in the spleen were analyzed by ELISA method. Cytokine levels in the spleen were divided according to the total protein levels. Values are the mean ± SEM. FPP: fermented porcine placenta, Lys: lysine, Leu: leucine. *p< 0.05, significantly different from the control mice.
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