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首页> 中文期刊> 《动物营养学报》 >防御素基因在乌金猪与约大乌猪组织中的表达及东亚飞蝗抗菌活性物质对防御素基因表达的调节作用

防御素基因在乌金猪与约大乌猪组织中的表达及东亚飞蝗抗菌活性物质对防御素基因表达的调节作用

         
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摘要

This experiment was aimed to investigate the expression of β⁃defensin⁃1 ( pBD⁃1) , β⁃defensin⁃2 ( pBD⁃2) and β⁃defensin⁃3 ( pBD⁃3) genes in different tissues of Wujin pigs and Yuedawu pigs with the blood of Wujin pigs, and also investigate the effect of locust antibacterial active substances ( LAAS) on the immune stress parameters and expression of β⁃defensins ( pBDs) in procine fetal skin fibroblasts ( PFSF) . The expres⁃sion differences of pBD⁃1, pBD⁃2 and pBD⁃3 genes were determined by real⁃time PCR in 12 tissues such as heart, liver, lung, kidney, jejunum, ileum, duodenum, skin, muscle, pancreas, testis and ovary of 2⁃month⁃old Wujin and Yuedawu pigs. The cultured 4th to 5th generations of PFSF were used, and immune stress model was induced by lipopolysaccharides (LPS). Adding different concentrations (0, 75, 150, 300, 600 and 1 200 μg/mL) of LAAS in DMEM/F12 culture medium, and the effect of LAAS on the immune stress pa⁃rameters and expression of pBD⁃1, pBD⁃2 and pBD⁃3 genes in PFSF were studied. The results showed as fol⁃lows:1) the serum lysozyme ( LSZ) activity and immunoglobulin G ( IgG) content of Wujin pigs were signif⁃icantly higher than those of Yuedawu pigs ( P<0.05) , and serum nitric oxide ( NO) content of Wujin pigs was extremely significantly lower than that of Yuedawu pigs ( P<0.01) . 2) The expression of pBD⁃1, pBD⁃2 and pBD⁃3 genes in most tissues of Yuedawu pigs were higher than those of Wujin pigs. The expression of pBD⁃1 and pBD⁃3 genes in skin and ovary of two breeds pigs were higher than those in other tissues, and the expres⁃sion of pBD⁃2 gene in liver and kidney of two breeds pigs was higher than that in other tissues. 3) The supple⁃mentation of 300 μg/mL LAAS extremely significantly decreased lactate dehydrogenase ( LDH ) activity in culture medium (P<0.01). LAAS could extremely significantly increased NO content in culture medium ex⁃cept for the 75μg/mL LAAS group in normal cells ( P<0.01) , and that in normal cells and stressed cells both reached the highest when LAAS concentration was 600 μg/mL. When LAAS concentrations were 0, 75 and 150 μg/mL, nitric oxide synthase ( NOS) activity in stressed cells was significantly higher than that in normal cells (P<0.05 or P<0.01). The supplementation of 1 200 μg/mL LAAS extremely significantly decreased LSZ activity in culture medium ( P < 0. 01 ) . When LAAS concentrations were 150, 300, 600 and 1 200 μg/mL, LSZ activity had significant differences in normal cells and stressed cells ( P<0. 05 or P<0.01) . 4) The expression of pBD⁃1, pBD⁃2 and pBD⁃3 genes in stressed cells were higher than those in nor⁃mal cells. When LAAS concentration was 150 μg/mL, the expression of pBD⁃1, pBD⁃2 and pBD⁃3 genes all reached the highest. The results indicate that there are significant differences in species and tissues specificity of the expression of pBDs genes in Wujin and Yuedawu pigs. The proper concentration of LAAS can decrease LDH activity, but increase NO content and the activities of NOS and LSZ in PFSF, and up⁃regulate the expres⁃sion of pBDs in both stressed cells and normal cells.%本试验旨在检测β防御素－1（ pBD⁃1）、β防御素－2（ pBD⁃2）和β防御素－3（ pBD⁃3）基因在乌金猪和具有乌金猪血缘的约大乌猪不同组织中的表达，并探讨东亚飞蝗抗菌活性物质（ LAAS）对猪胎儿皮肤成纤维细胞（ PFSF）免疫应激参数和防御素（ pBDs）基因表达的调节作用。采用实时荧光定量PCR方法检测2月龄乌金猪与约大乌猪心脏、肝脏、肺脏、肾脏、空肠、回肠、十二指肠、皮肤、肌肉、胰腺、睾丸或卵巢这12种组织中pBD⁃1、pBD⁃2和pBD⁃3基因的表达差异；采用培养至第4～5代的PFSF，通过脂多糖（ LPS）诱导建立免疫应激模型，在DMEM／F12培养液中添加不同浓度（0、75、150、300、600和1200μg／mL ）的LAAS，考察LAAS对PFSF免疫应激参数和pBD⁃1、pBD⁃2、pBD⁃3基因表达的影响。结果表明：1）乌金猪血清溶菌酶（ LSZ ）活性、免疫球蛋白G（ IgG）含量显著高于约大乌猪（ P＜0．05），血清一氧化氮（ NO）含量极显著低于约大乌猪（ P＜0．01）。2）约大乌猪大多数组织中pBD⁃1、pBD⁃2和pBD⁃3基因的表达量高于乌金猪；2个品种猪的pBD⁃1和pBD⁃3基因均在皮肤和卵巢中表达量较高，pBD⁃2基因在肝脏和肾脏中表达量较高。3）300μg／mL LAAS极显著降低了培养液中乳酸脱氢酶（ LDH ）活性（ P＜0．01）；LAAS极显著增加了培养液中NO含量（除75μg／mL组正常细胞外）（ P＜0．01），正常细胞和应激细胞均在LAAS浓度为600μg／mL时达到最高；LAAS浓度为0、75和150μg／mL时应激细胞一氧化氮合酶（NOS）活性显著或极显著高于正常细胞（P＜0．05或 P＜0．01）；添加1200μg／mL LAAS可极显著降低培养液中的LSZ活性（ P＜0．01），LAAS浓度为150、300、600和1200μg／mL时应激细胞与正常细胞中LSZ 活性差异显著或极显著（ P＜0．05或P＜0．01）。4）应激细胞pBD⁃1、pBD⁃2和pBD⁃3基因的表达量均高于正常细胞；当LAAS浓度为150μg／mL时，pBD⁃1、pBD⁃2和pBD⁃3基因的表达量最高。结果提示，乌金猪和约大乌猪防御素基因的表达存在明显的品种差异性和组织特异性，适宜浓度的LAAS能够降低PFSF中LDH活性，提高NO含量及NOS、LSZ活性，上调应激细胞与正常细胞防御素的表达量。

著录项

  • 来源
    《动物营养学报》 |2016年第7期|2096-2105|共10页
  • 作者

    李美荃; 刘红; 张春勇; 安清聪; 骆雪; 陈克嶙; 郭荣富;

  • 作者单位

    云南农业大学;

    云南省动物营养与饲料重点实验室;

    昆明 650201;

    云南农业大学;

    云南省动物营养与饲料重点实验室;

    昆明 650201;

    云南农业大学;

    云南省动物营养与饲料重点实验室;

    昆明 650201;

    云南农业大学;

    云南省动物营养与饲料重点实验室;

    昆明 650201;

    云南农业大学;

    云南省动物营养与饲料重点实验室;

    昆明 650201;

    云南农业大学;

    云南省动物营养与饲料重点实验室;

    昆明 650201;

    云南农业大学;

    云南省动物营养与饲料重点实验室;

    昆明 650201;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 猪;
  • 关键词

    乌金猪; 约大乌猪; β-防御素; 免疫应激; 东亚飞蝗抗菌活性物质;

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