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首页> 外文期刊>Frontiers in Cellular and Infection Microbiology >Nocardia cyriacigeogica from Bovine Mastitis Induced In vitro Apoptosis of Bovine Mammary Epithelial Cells via Activation of Mitochondrial-Caspase Pathway
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Nocardia cyriacigeogica from Bovine Mastitis Induced In vitro Apoptosis of Bovine Mammary Epithelial Cells via Activation of Mitochondrial-Caspase Pathway

机译：牛乳腺炎诺卡氏菌通过线粒体半胱天冬酶途径的激活诱导牛乳腺上皮细胞的体外凋亡

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Nocardia is one of the causing agents of bovine mastitis and increasing prevalence of nocardial mastitis in shape of serious outbreaks has been reported from many countries. However, the mechanisms by which this pathogen damages the bovine mammary epithelial cells (bMECs) is not yet studied. Therefore, this study was designed with the aim to evaluate the apoptotic effects elicited by Nocardia and to investigate the pathway by which the Nocardia induce apoptosis in bMECs. Clinical N. cyriacigeorgica strain from bovine mastitis was used to infect the bMECs for different time intervals, viz. 1 h, 3 h, 6 h, 12 h, and 18 h, and then the induced effects on bMECs were studied using adhesion and invasion assays, release of lactate dehydrogenase (LDH), apoptosis analysis by annexin V and propidium iodide (PI) double staining, morphological and ultrastructural observations under scanning electron microscope (SEM) and transmission electron microscope (TEM), mitochondrial transmembrane potential (ΔΨm) assay using flow cytometry, and the protein quantification of mitochondrial cytochrome c and caspase-9 and caspase-3 by western blotting. The results of this study showed that N. cyriacigeorgica possessed the abilities of adhesion and invasion to bMECs. N. cyriacigeorgica was found to collapse mitochondrial transmembrane potential, significantly (p 0.05) release mitochondrial cytochrome c and ultimately induce cell apoptosis. Additionally, it promoted casepase-9 (p 0.01) and casepase-3 (p 0.05) levels, significantly (p 0.01) increased the release of LDH and promoted DNA fragmentation which further confirmed the apoptosis. Furthermore, N. cyriacigeorgica induced apoptosisecrosis manifested specific ultrastructure features under TEM, such as swollen endoplasmic reticulum, cristae degeneration and swelling of mitochondria, vesicle formation on the cell surface, rupturing of cell membrane and nuclear membrane, clumping, fragmentation and margination of chromatin. The present study is the first comprehensive insight into patho-morphological ultrastructural features of apoptosisecrosis induced by N. cyriacigeorgica, which concluded that the clinical N. cyriacigeorgica induced apoptotic changes in the bMECs through mitochondrial-caspase dependent apoptotic pathway.
机译：诺卡氏菌是牛乳腺炎的病原体之一，许多国家已经报道，以严重爆发的形式出现的诺卡氏乳腺炎患病率增加。但是，尚未研究这种病原体损害牛乳腺上皮细胞（bMECs）的机制。因此，本研究旨在评估诺卡氏菌引起的凋亡作用，并研究诺卡氏菌诱导bMECs凋亡的途径。来自牛乳腺炎的临床N. cyriacigeorgica菌株用于感染bMEC不同时间间隔，即。 1 h，3 h，6 h，12 h和18 h，然后通过粘附和侵袭试验，乳酸脱氢酶（LDH）释放，膜联蛋白V和碘化丙啶（PI）的凋亡分析研究对bMECs的诱导作用。在扫描电子显微镜（SEM）和透射电子显微镜（TEM）下进行双重染色，形态学和超微结构观察，使用流式细胞仪检测线粒体跨膜电位（ΔΨm），并通过线粒体细胞色素c和caspase-9和caspase-3定量蛋白质蛋白质印迹。这项研究的结果表明，N。cyriacigeorgica具有对bMECs的粘附和侵袭能力。发现N. cyriacigeorgica破坏了线粒体跨膜电位，显着（p <0.05）释放线粒体细胞色素c，并最终诱导细胞凋亡。此外，它促进了casepase-9（p <0.01）和casepase-3（p <0.05）的水平，显着（p <0.01）增加了LDH的释放并促进了DNA片段化，进一步证实了细胞凋亡。此外，cyriacigeorgica诱导的凋亡/坏死在TEM下表现出特定的超微结构特征，例如内质网肿胀，线粒体的cr退化和肿胀，细胞表面的囊泡形成，细胞膜和核膜破裂，结块，破碎和边缘化染色质本研究是对cy.ci.ciorgorgica诱导的凋亡/坏死的病理形态超微结构特征的首次全面了解，其结论是临床的cy.ci.cigeorgorgica通过线粒体-胱天蛋白酶依赖性凋亡途径诱导了bMECs的凋亡变化。

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