摘要
转化生长因子β1受体(transforming growth factor-β 1,TGF-β1)是CD14单核细胞中的生存因子,当TGF-β1缺失时CD14单核细胞迅速执行凋亡程序。然而,目前有关TGF-β1基因在绵羊乳腺炎发生过程中的调控机制尚不清楚。为研究TGF-β1基因在患临床型乳房炎绵羊乳腺组织中的表达差异,本试验使用NCBI网站已公布的绵羊TGF-β1基因CDS序列,运用生物信息学分析方法对该基因生物学特征进行预测。通过qRT-PCR、Western blot检测健康和临床型乳腺炎绵羊乳腺组织中TGF-β1 mRNA及蛋白的表达,并使用免疫组织化学染色确定该蛋白在绵羊乳腺组织内分布情况。生物信息学分析结果表明,绵羊TGF-β1蛋白为疏水性不稳定蛋白,属于分泌蛋白,不同物种间保守性较高。qRT-PCR与蛋白免疫印迹结果显示,在健康和临床型乳腺炎乳腺组织中均存在TGF-β1基因的表达,且TGF-β1 mRNA与蛋白相对表达量均极显著(P<0.01)高于健康组。免疫组织化学结果显示,TGF-β1蛋白在临床型乳腺炎乳腺组织中均强烈表达于基质细胞和乳腺上皮细胞。由此推测可能是由于TGF-β1基因表达上调,导致患临床型乳腺炎乳腺组织中基质细胞与乳腺上皮细胞中炎症反应加剧。这为进一步研究TGF-β1基因在绵羊乳腺炎发病机制中的调控作用提供基础信息。
Transforming growth factor-β1(TGF-β1)receptor is a survival factor in CD14 monocytes,and CD14 monocytes rapidly perform apoptotic program when TGF-β1 is depleted.However,it is not clear about the regulatory mechanism of TGF-β1 gene in the development of mastitis in sheep.To investigate the differential expression of the TGF-β1 gene in mammary tissue of sheep with clinical mastitis,this experiment used the published CDS sequence of the sheep TGF-β1 gene from the NCBI website to predict the biological characteristics of the gene using bioinformatics analysis methods.The expression of TGF-β1 mRNA and protein in mammary tissues of healthy and clinically mastitic sheep was measured by qRT-PCR and Western blot,and the distribution of the protein in sheep mammary tissues was determined by immunohistochemical staining.Bioinformatics analysis showed that sheep TGF-β1 protein is a hydrophobic unstable protein,a secreted protein,and highly conserved among different species.Moreover,qRT-PCR and western blot showed that TGF-β1 gene was expressed in both healthy and clinical mastitis mammary tissues,and the relative expression levels of TGF-β1 mRNA and protein were extremely significantly(P<0.01)higher than healthy group.Immunohistochemical results showed that TGF-β1 protein was strongly expressed in stromal cells and mammary epithelial cells in clinical mastitis breast tissue.It was hypothesized that the up-regulation of TGF-β1 gene expression may lead to increased inflammatory response in stromal cells and mammary epithelial cells in mammary tissue with clinical mastitis.These results provided basic information for further research on the regulatory role of TGF-β1 gene in the pathogenesis of ovine mastitis.
作者
马克岩
尹德恩
马友记
MA Keyan;YIN Deen;MA Youji(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation,Lanzhou 730070,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2023年第7期1471-1477,共7页
Chinese Journal of Veterinary Science
基金
甘肃省引导科技创新专项资助项目(2019ZX-07)。
