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Effects of maternal l-proline supplementation on inflammatory cytokines at the placenta and fetus interface of mice

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Abstract

Dietary l-proline (proline) supplementation during gestation enhances fetal survival and placental development in mice. The objective of the present study was to test the hypothesis that this beneficial effect of proline was associated with alterations in inflammatory response at the placenta and fetus interface. Populations of immune cells present in peripheral blood mononuclear cells (PBMC) were determined by flow cytometry analysis. The concentrations of immunoglobulins in plasma, and the concentrations of cytokines in plasma, uterus, placenta, and amniotic fluid were measured using a bead-based immunoassay. The data showed that proline supplementation led to higher (P < 0.05) populations of B lymphocytes (CD3−CD19+), natural killer (NK) cells (CD3−NK1.1+), and dendritic cells (DCs, CD11c+MHCII+) in peripheral blood, as compared with the controls. Conversely, mice fed a proline-supplemented diet had a lower population of neutrophils (CD11b+F4/80−). Further study showed that proline supplementation decreased (P < 0.05) the concentrations of (1) interleukin (IL)-23, IL-1α, and IL-6 in plasma; (2) IL-6 in the uterus; and (3) tumor necrosis factor alpha (TNF-α), monocyte chemotactic protein (MCP)-1, and IL-17 in the placenta; and (4) interferon (IFN)-γ in amniotic fluid, compared with controls. Conversely, proline supplementation resulted in higher (P < 0.05) concentrations of (1) IL-10, IL-17 and granulocyte–macrophage colony-stimulating factor (GM-CSF) in plasma; (2) IL-10 and IL-1α in the uterus; and (3) IL-1α, IL-1β, IL-10, IL-27, and IFN-β in amniotic fluid, compared with controls. Moreover, concentrations of immunoglobulin (Ig) G2b and IgM were enhanced (P < 0.05) by proline administration. Taken together, our results reveal a regulatory effect of proline in the immunological response at the maternal–fetal interface, which is critical for embryonic development and fetal survival.

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Abbreviations

APC:

Allophycocyanin conjugated

DCs:

Dendritic cells

E:

Embryonic day

FITC:

Fluorescein isothiocyanate-conjugated

GM-CSF:

Granulocyte–macrophage colony-stimulating factor

IFN:

Interferon

Ig:

Immunoglobulin

IL:

Interleukin

MCP-1:

Monocyte chemotactic protein 1

MHC:

Major histocompatibility complex

NK:

Natural killer

PBMC:

Peripheral blood mononuclear cells

PE:

Phycoerythrin

PerCP:

Peridinin chlorophyll protein complex-conjugated

Proline:

L-Proline

RBC:

Red blood cell

TNF-α:

Tumor necrosis factor alpha

Th:

T helper

Treg:

Regulatory T

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 31572410, 31572412, 31625025), the Zhengzhou 1125 Talent Program, Jinxinnong Animal Science Development Foundation, 111 project (B16044), and Texas A&M AgriLife Research (H-8200).

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Authors and Affiliations

  1. Department of Animal Nutrition and Feed Science, State Key Laboratory of Animal Nutrition, China Agricultural University, Beijing, 100193, China

    Ning Liu, Jingqing Chen, Yu He, Hai Jia, Da Jiang, Shuai Li, Ying Yang, Zhaolai Dai, Zhenlong Wu & Guoyao Wu

  2. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China

    Ning Liu

  3. Department of Animal Science, Texas A&M University, College Station, TX, 77843, USA

    Guoyao Wu

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  1. Ning Liu
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Liu, N., Chen, J., He, Y. et al. Effects of maternal l-proline supplementation on inflammatory cytokines at the placenta and fetus interface of mice. Amino Acids 52, 587–596 (2020). https://doi.org/10.1007/s00726-020-02837-0

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