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Expression of cathelicidins mRNA in the goat mammary gland and effect of the intramammary infusion of lipopolysaccharide on milk cathelicidin-2 concentration

机译:cathelicidins mRNA在山羊乳腺中的表达及乳内灌注脂多糖对牛奶cathelicidin-2浓度的影响

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Cathelicidins are a family of antimicrobial peptides found in neutrophils and the epithelium that have broad-spectrum activity against bacteria. This study aimed to investigative the mRNA expression of cathelicidins and protein localization of cathelicidin-2 in the goat mammary gland and its secretion into milk. The mRNA expression of cathelicidins was examined in different regions of the mammary gland by reverse transcription PCR. A cathelicidin-2 antibody was raised in rabbits by immunization with a synthetic cathelicidin-2 peptide consisting of 17 amino acids. The protein localization of cathelicidin-2 was investigated in the mammary gland by immunohistochemistry. Skim milk was collected before (0 h) and 2, 4, 8, 12, and 24 h after the intramammary infusion of lipopolysaccharide and saline, and the concentration of cathelicidin-2 was examined by an enzyme immunoassay. The mRNAs of cathelicidin-1, 2, 3, 6, and 7 were expressed in both the teat and deep region of the mammary gland from healthy and mastitic goats. Immunoreactive cathelicidin-2 was not localized in the epithelial cells of the teat skin, teat cistern, or mammary alveoli, whereas it was localized in polymorphonuclear cells in the mammary gland and those collected from the blood and milk. Cathelicidin-2 was detected in skim milk by Western blotting. The concentration of cathelicidin-2 in milk increased 4 h after the intramammary infusion of lipopolysaccharide. These results suggest that cathelicidin-2 is expressed in polymorphonuclear cells and is secreted into milk in goat. (c) 2014 Elsevier B.V. All rights reserved.
机译:鞘磷脂是在嗜中性粒细胞和上皮细胞中发现的抗菌肽家族,对细菌具有广谱活性。这项研究旨在调查山羊乳腺中cathelicidins的mRNA表达和cathelicidin-2的蛋白质定位及其分泌到乳汁中的情况。通过逆转录PCR检查了乳腺不同区域中cathelicidins的mRNA表达。通过用由17个氨基酸组成的合成cathelicidin-2肽进行免疫,可在兔中产生cathelicidin-2抗体。通过免疫组织化学研究了cathelicidin-2在乳腺中的蛋白质定位。在乳房内输注脂多糖和盐水之前(0小时)和2、4、8、12和24小时之前收集脱脂奶,并通过酶免疫法检测cathelicidin-2的浓度。 cathelicidin-1、2、3、6和7的mRNA在健康和乳香山羊的乳头和乳腺深处均有表达。免疫反应性cathelicidin-2不在乳头皮肤,乳头水箱或乳腺泡的上皮细胞中定位,而在乳腺的多形核细胞中以及从血液和牛奶中收集的细胞中则没有定位。通过蛋白质印迹法在脱脂牛奶中检测到Cathelicidin-2。乳腺内注入脂多糖后4小时,牛奶中cathelicidin-2的浓度增加。这些结果表明,cathelicidin-2在多形核细胞中表达,并分泌到山羊的牛奶中。 (c)2014 Elsevier B.V.保留所有权利。

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