SOCE调控亚临床低钙血症奶牛单核巨噬细胞吞噬和黏附功能的研究

doi:10.16431/j.cnki.1671-7236.2025.08.039

Abstract

Abstract: 【Objective】 The purpose of this experiment was to explore the effect of store operated Ca^2＋ entry (SOCE) on the phagocytic and adhesion functions of mononuclear macrophages in dairy cows. 【Method】 Peripheral blood of healthy (control，Con)and subclinical hypocalcemia dairy cows (hypocalcemia,Hyp) was collected,and CD14^＋ mononuclear macrophages in the blood were isolated and cultured in vitro using magnetic beads.The mononuclear macrophages of subclinical hypocalcemia dairy cows were divided into three groups: Hyp, thapsigarsin (TG) treatment group (Hyp+TG) and 2-aminoethyl diphenylborinate (2-APB) treatment group (Hyp＋2-APB).The intracellular Ca^2＋ level was detected by flow cytometry.Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expression of SOCE-related factors.The phagocytosis and adhesion of mononuclear macrophages were detected by laser confocal and immunofluorescence,and the expression of phagocytosis and adhesion factors of mononuclear macrophages was detected. 【Result】 Flow cytometry analysis showed that,compared with Con group,the intracellular Ca^2＋ level of mononuclear macrophages in Hyp group was extremely significantly decreased (P＜0.01).Real-time quantitative PCR results showed that,compared with Con group,the expression of calcium release-activated calcium modulator 1 (ORAI1)，stromal interaction molecule 1 (STIM1),STIM2 and selectin L (SELL) genes of mononuclear macrophages in Hyp group were extremely significantly decreased (P＜0.01)，expression of ORAI2,ORAI3 and myeloperoxidase (MPO) genes were significantly decreased (P＜0.05).Western blotting results showed that,compared with Con group,the expression of ORAI2,ORAI3,STIM2,ORAI1 and STIM1 proteins of mononuclear macrophages in Hyp group were extremely significant or significantly decreased (P＜0.01 or P＜0.05).Meanwhile,compared with Con group,the phagocytic and adhesion abilities of mononuclear macrophages in Hyp group were significantly or extremely significantly decreased (P＜0.05 or P＜0.01).After TG treatment,compared with Hyp group,the expression of ORAI1,ORAI2,MPO,ORAI3,STIM1,STIM2 and SELL genes in mononuclear macrophages of subclinical hypocalcemia dairy cows were significantly or extremely significantly increased (P＜0.05 or P＜0.01)，the expression of ORAI1,ORAI3,STIM1,STIM2 and ORAI2 proteins were significantly or extremely significantly upregulated (P＜0.05 or P＜0.01).The phagocytic and adhesion abilities of cells were significantly or extremely significantly enhanced (P＜0.05 or P＜0.01).After 2-APB treatment,compared with Hyp group,the expression of ORAI1,ORAI2,MPO,ORAI3,STIM1,STIM2 and SELL genes in mononuclear macrophages of subclinical hypocalcemia dairy cows were significantly or extremely significantly decreased (P＜0.05 or P＜0.01)，the expression of ORAI2,STIM2,ORAI1,ORAI3 and STIM1 proteins were significantly or extremely significantly decreased (P＜0.05 or P＜0.01).The phagocytic and adhesion abilities of cells were extremely significant or significantly weakened (P＜0.01 or P＜0.05). 【Conclusion】 After SOCE was activated by TG,the Ca^2＋ level in mononuclear macrophages of subclinical hypocalcemia dairy cows increased,and the phagocytic and adhesion abilities enhanced.After SOCE was inhibited by 2-APB,the intracellular Ca^2＋ level in mononuclear macrophages of subclinical hypocalcemia dairy cows decreased,and the phagocytic and adhesion functions weakened，which indicated that SOCE was involved in regulating the phagocytosis and adhesion of mononuclear macrophages.

Key words: store operated Ca^2＋ entry (SOCE); mononuclear macrophage; phagocytic; adhesion

CLC Number:

S856

LI Siyao, JI Ziwei, LI Lei, ZHANG Jingqi, YUE Baichen, YIN Yufeng, XU Chuang, YANG Wei, ZHANG Bingbing. Regulation of Phagocytic and Adhesion Functions of Mononuclear Macrophages by SOCE in Subclinical Hypocalcemia Dairy Cows[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3907-3917.

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Regulation of Phagocytic and Adhesion Functions of Mononuclear Macrophages by SOCE in Subclinical Hypocalcemia Dairy Cows

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