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Int J Mol Sci
2017 Oct 08;1810:. doi: 10.3390/ijms18102110.
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Peptides from Colochirus robustus Enhance Immune Function via Activating CD3ζ- and ZAP-70-Mediated Signaling in C57BL/6 Mice.
Du X
,
Lian F
,
Li Y
,
Li D
,
Wu D
,
Feng Q
,
Feng Z
,
Li Y
,
Bu G
,
Meng F
,
Cao X
,
Chen Z
,
Zeng X
.
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Colochirus robustus, a species of sea cucumber, has long been used in East and Southeast Asia as nutritious food as well as for certain medicinal purpose. Studies have shown a number of biological functions associated with consumption of sea cucumber, many of which are attributed to its major component, sea cucumber peptides (SCP). However, how SCP impacts immune system, which is critical for host defense, has not been defined. To address this issue, in the present study, we conducted comprehensive analysis of immune function after oral administration of SCP (0, 25, 50, and 75 mg/kg body weigh) for eight weeks in C57BL/6 mice. We found that SCP treatment significantly enhanced lymphocyte proliferation, serum albumin (ALB) levels, and the natural killer (NK) cell activity. Moreover, SCP promoted functions of helper T cells (Th) as indicated by increased production of Th1 type cytokines of Interleukin (IL)-1β, IL-2, Interferon (IFN)-γ and TNF-α and Th2 type cytokines (IL-4, IL-6, and IL-10). To determine the effective components, SCP was hydrolyzed into 16 types of constituent amino acids in simulated gastrointestinal digestion and these hydrolytic amino acids (HAA) were used for the mechanistic studies in the in vitro models. Results showed that HAA enhanced lymphocyte proliferation and production of IL-2, IL-10 and IFN-γ. Furthermore, CD3ζ (CD3ζ) and ζ-chain-associated protein kinase 70 (ZAP-70), the signaling molecules essential for activating T lymphocytes, were significantly up-regulated after HAA treatment. In summary, our results suggest that SCP is effective in enhancing immune function by activating T cells via impacting CD3ζ- and ZAP-70-mediated signaling pathway.
Figure 7. Effect of in vitro HAA supplementation on expression of: CD3ζ (A) and ζ -chain-associated protein kinase 70 (ZAP-70) (B) Splenocytes were incubated in the presence of HAA at 0, 0.25, 0.5 or 1 mg/mL for 4 h and then stimulated by CD3/CD28 for 48 h. CD3ζ and ZAP-70 expression was determined using flow cytometry; (A,B) Statistical summary of CD3ζ and ZAP-70 expression presented as mean fluorescence intensity (MFI), respectively; (C,D) Representative histograms for CD3ζ and ZAP-70, respectively. Values are means ± SD, n = 10. Means in a row without a common letter significantly differ as determined by one-factor ANOVA, p < 0.05.
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