动物营养学报2016，Vol.28Issue(7)：2096-2105,10.DOI:10.3969/j.issn.1006-267x.2016.07.014
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
The Expression of β-Defensins Genes in Tissues of Wujin and Yuedawu Pigs and Effect of Locust Antibacterial Active Substances on Expression of β-Defensins Genes
摘要
Abstract
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.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Key words
Wujin pigs/Yuedawu pigs/β-defensins/immune stress/locust antibacterial active substances分类
畜牧业引用本文复制引用
李美荃,刘红,张春勇,安清聪,骆雪,陈克嶙,郭荣富..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[J].动物营养学报,2016,28(7):2096-2105,10.基金项目
云南省重大科技专项基金（2012ZA018）；云南现代农业生猪产业技术体系2014［云财教］105＃、2015［云财教］90＃、2016［云财教］112＃ （）
