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首页> 美国卫生研究院文献>Journal of Animal Science >PSVI-4 Modulation of galectin gene expression and secretion in cow blood stimulated with Phorbol 12-myristate 13-acetate
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PSVI-4 Modulation of galectin gene expression and secretion in cow blood stimulated with Phorbol 12-myristate 13-acetate

机译：PSVI-4在牛血液中的Galectin基因表达和分泌的调节用Phorbol 12-Myristerate 13-乙酸酯刺激

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The objective of this study was to assess the expression and secretion of galectin gene in cow blood stimulated with Phorbol 12-myristate 13-acetate (PMA). Phorbol 12-myristate 13-acetate is a phorbol diester that activates the enzyme protein kinase C (PKC). Activated PKC is associated with regulation of inflammation. Galectins (Gal) are β-galactoside binding proteins that modulate inflammation following intra and extra-cellular secretion. Blood was collected from the jugular vein of clinically healthy Holstein-Friesian cows (n = 3) from North Carolina A&T diary unit. Individual samples were treated with either 10ng/ml of PMA or Phosphate buffer saline (PBS) for 30 minutes at 370 C. Total RNA was isolated from blood, pooled and transcribed to cDNA, for real-time PCR. Cow specific primers LGALS -1, -2, -3, -4, -7, -8, -9, -12 were designed using the NCBI Primer 3 tool. Housekeeping genes RPLP0 and UCHL5 were used for normalization. Fold change was calculated using the Livak method (Fold change >2 were considered as significant). Secretion of GAL -1, -2, -3, -4, -8, -9 was assessed using bovine GAL specific ELISA kits. Data were analyzed by one-way ANOVA using SAS version 9.4. Treatment with PMA up regulated the transcription of LGALS -2, -4, -8, (2.28, 2.36, 2.39 folds respectively) and down regulated the transcription of LGALS -9 (-5.04 fold). Transcription of LGALS -1, -3, -7, 12 remained unchanged. All tested GAL were secreted. Treatment with PMA increased the secretion level of GAL -1, -3, -4 and decreased the secretion level of GAL -2, -4, -9 (P > 0.05). Thus, activation of PKC differentially modulates galectin gene expression and secretion in cow blood. The observed differences in treatment and control groups warrant further studies.
机译：本研究的目的是评估母牛血液中的半乳糖型基因的表达和分泌，刺激磷血液13-醋酸酯（PMA）。 Phorbol 12-Myristate 13-醋酸酯是一种磷博二酯，可激活酶蛋白激酶C（PKC）。活化的PKC与炎症的调节有关。半乳糖素（GAL）是调节血液内和外细胞分泌后调节炎症的β-半乳糖苷结合蛋白。从北卡罗来纳A＆T日志单位从临床健康的Holstein-Friesian（n = 3）的颈静脉收集血液。用10ng / ml的PMA或磷酸盐缓冲盐（PBS）处理单个样品，在370℃下30分钟。从血液中分离总RNA，合并并转录到cDNA，用于实时PCR。使用NCBI底漆3工具设计母牛特异性引物LGALS -1，-2，-3，-4，-7，-8，-9，-12。家政基因RPLP0和UCHL5用于标准化。使用Livak方法计算折叠变化（折叠变化> 2被认为是显着的）。使用牛GAL特异性ELISA试剂盒评估GAL -1，-2，-3，-4，-8，-9的分泌。使用SAS版本9.4通过单向ANOVA进行分析数据。用PMA的处理调节LGALS -2，-4，-8，（分别分别为2.28,2.36,2.39倍的转录，下调LGALS -9（-5.04倍）的转录。 LGALS -1，-3，-7,12的转录保持不变。所有测试的Gal都分泌。用PMA处理增加了GAL -1，-3，-4的分泌水平，并降低了GAL-2，-4，-9（P> 0.05）的分泌水平。因此，PKC的激活差异地调节牛血液中的半乳糖基因表达和分泌。观察到的治疗和对照组的差异需要进一步研究。

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