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首页> 外文期刊>Journal of Molecular Biology Research >Phorbol 12-myristate 13-acetate (PMA) Alters Galectin Gene Expression in Cows
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Phorbol 12-myristate 13-acetate (PMA) Alters Galectin Gene Expression in Cows

机译：Phorbol 12-肉豆蔻酸酯13-醋酸酯（PMA）改变了奶牛的Galectin基因表达

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Galectins (GAL) are β-galactoside binding proteins. The objective of this study was to assess the effect of PMA on the regulation of galectin gene expression in bovine blood and neutrophils. Blood was collected from 3 clinically healthy Holstein cows. Neutrophils were isolated by differential centrifugation. Blood and neutrophils were treated with 10ng/ml of PMA or maintained in phosphate-buffered saline for 30 minutes at 370 C. Pooled total RNA were transcribed to cDNA for real-time PCR. Specific primers for bovine LGALS -1, -2, -3, -4, -7, -8, -9, -11, -12, and their binding proteins (bp) LGALS3bp, T cell immunoglobulin and mucin domain 3 (TIM-3) were used. Housekeeping genes RPLP0 and UCHL5 served as internal controls. Fold changes were calculated using the Livak method. Total protein concentration was assessed using the Bicinchoninic acid assay. Secretion of GAL -1, -2, -3, -4, -8, -9 was assessed using bovine GAL specific ELISA. All tested LGALS were detected. Activation with PMA differentially modulated expression and secretion of GAL in blood and neutrophils. Expression levels of LGALS -1 and LGALS -3 was not affected. Expression of LGALS -7, -8, -9, -11, -12, and TIM-3 was differentially modulated. The secretion of GAL -3 was significantly increased in response to PMA. The most pronounced effect of PMA was observed on LGALS-9, TIM-3 expression, and the secretion of GAL -3. Thus, GAL may serve as biomarkers of cell activation with functional consequences and warrant further study.
机译：半乳糖素（GAL）是β-半乳糖糖苷结合蛋白。本研究的目的是评估PMA对牛血液和中性粒细胞的Galectin基因表达调控的影响。从3个临床健康的Holstein奶牛收集血液。通过差动离心分离中性粒细胞。用10ng / ml PMA处理血液和中性粒细胞，或者在370℃下在磷酸盐缓冲的盐水中保持30分钟。合并的总RNA转录到CDNA以进行实时PCR。牛LGALS -1，-2，-3，-4，-7，-8，-9，-11，-12和它们的结合蛋白（bp）Lgals3bp，t细胞免疫球蛋白和粘蛋白结构蛋白3的特异性引物（inch使用-3）。家政基因RPLP0和UCHL5作为内部控制。使用Livak方法计算折叠变化。使用双式胆酸测定评估总蛋白质浓度。使用牛GAL特异性ELISA评估GAL -1，-2，-3，-4，-8，-9的分泌。检测到所有测试的LGALS。用PMA差异调节血液和中性粒细胞的PMA差异调节表达和分泌。 LGALS -1和LGALS -3的表达水平不受影响。 LGALS -7，-8，-9，-11，-12和TIM-3的表达被差异调节。响应于PMA，GAL -3的分泌显着增加。在LGALS-9，TIM-3表达和GAL -3的分泌中观察到PMA最明显的效果。因此，GAL可以用作细胞活化的生物标志物，具有功能后果和担心进一步研究。

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