Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytes  

作　　者：Laman K.Mamedova Kirby C.Krogstad Paiton O.McDonald Laxman Pokhrel Duy H.Hua Evan C.Titgemeyer Barry J.Bradford 

机构地区：[1]Department of Animal Science,Michigan State University,East Lansing,Michigan 48824,USA [2]Department of Animal Sciences and Industry,Kansas State University,Manhattan,KS 66506,USA [3]Comparative Medicine and Integrative Biology,East Lansing,MI 48824,USA [4]Department of Chemistry,Kansas State University,Manhattan,KS 66506,USA

出　　处：《Journal of Animal Science and Biotechnology》2024年第4期1567-1577,共11页畜牧与生物技术杂志（英文版）

基　　金：the National Institutes of Health, National Institute of General Medical Sciences (R01 GM128659) for financial support;The Attune Cytpix, located in the MSU Flow Cytometry Core Facility, is supported by the Equipment Grants Program, award #2022-70410-38419, from the U.S. Department of Agriculture (USDA), National Institute of Food and Agriculture (NIFA)。

摘　　要：Background Dairy cows experiencing ketosis after calving suffer greater disease incidence and are at greater risk of leaving the herd. In vitro administration of beta-hydroxybutyric acid(BHBA;the primary blood ketone) has inhibitory effects on the function of bovine leukocytes. BHBA is a ligand of HCAR2 and the activation of these receptors promotes an anti-inflammatory response which may be related with immunosuppression observed in transition dairy cattle. The objective of this study was to identify and test antagonists for HCAR2 in bovine immune cells cultured with BHBA.Results We observed expression of HCAR2 at the protein level within lymphocytes, monocytes, and granulocytes. The proportion of cells expressing HCAR2 tended to be greater in mid-lactation compared to early lactation cows;the increase was a result of increased proportion of T and B cells expressing HCAR2. Stimulation of HCAR2 with niacin or BHBA promoted Ca^(2+) mobilization in neutrophils and mononuclear cells. Mononuclear cells treated with BHBA had diminished intracellular Ca^(2+) responses when HCAR2 was knocked down by si RNA silencing, indicating Ca^(2+) mobilization was mediated by HCAR2 signaling. Two candidate antagonists for HCAR2, synthesized from niacin(NA-1 and NA-5), were tested;monocytes and neutrophils pre-treated with NA-1 and NA-5 had reduced Ca^(2+) mobilization after incubation with BHBA. Furthermore, NA-5 but not NA-1 prevented BHBA-associated reductions in cyclic AMP.Conclusions We demonstrated that HCAR2 is present on bovine leukocytes and has greater expression later in lactation. We confirmed that BHBA and niacin derived HCAR2 antagonists alter bovine leukocyte activity. Our results demonstrate that both BHBA and niacin affect bovine leukocyte Ca^(2+) mobilization in a HCAR2-dependent manner.

关 键 词：Flow cytometry GPR109A HM74A KETOSIS 

分 类 号：S823[农业科学—畜牧学]