Shahid Muhammad, Gao Jian, Zhou Yanan, Liu Gang, Ali Tariq, Deng Youtian, Sabir Naveed, Su Jingliang, Han Bo
College of Veterinary Medicine, China Agricultural University, Beijing 100193, P R China.
Oncotarget. 2017 May 9;8(19):31938-31947. doi: 10.18632/oncotarget.16653.
Bovine protothecal mastitis results in considerable economic losses worldwide. However, Prototheca zopfii induced morphological alterations and oxidative stress in bovine mammary epithelial cells (bMECs) is not comprehensively studied yet. Therefore, the aim of this current study was to investigate the P. zopfii induced pathomorphological changes, oxidative stress and apoptosis in bMECs. Oxidative stress was assessed by evaluating catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), malondialdehyde (MDA) contents and lactate dehydrogenase (LDH) activity, while ROS generation and apoptosis was measured by confocal laser scanning microscopy. The results revealed that infection of P. zopfii genotype II (GTII) significantly changed bMECs morphology, increased apoptotic rate and MDA contents at 12 h (p < 0.05) and 24 h (p < 0.01) in comparison with control group, in time-dependent manner. LDH activity and ROS generation was also increased (p < 0.01) at 12 h and 24 h. However, SOD and CAT contents in bMECs infected with GTII were decreased (p < 0.05) at 12 h, while GPx (p < 0.01), SOD (p < 0.05) and CAT (p < 0.01) levels were reduced at 24 h. In case of GTI, only CAT and GPx activities were significantly decreased when the duration prolonged to 24 h but lesser than GTII. This suggested that GTII has more devastating pathogenic effects in bMECs, and the findings of this study concluded that GTII induced apoptosis and oxidative stress in bMECs via the imbalance of oxidant and antioxidant defenses as well as the production of intracellular ROS.
牛原虫性乳腺炎在全球范围内造成了相当大的经济损失。然而,尚缺乏对zopfii原虫引起的牛乳腺上皮细胞(bMECs)形态学改变和氧化应激的全面研究。因此,本研究的目的是探讨zopfii原虫诱导的bMECs病理形态学变化、氧化应激和细胞凋亡。通过评估过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)、丙二醛(MDA)含量和乳酸脱氢酶(LDH)活性来评估氧化应激,同时通过共聚焦激光扫描显微镜测量活性氧(ROS)生成和细胞凋亡。结果显示,与对照组相比,II型基因型(GTII)的zopfii原虫感染在12小时(p<0.05)和24小时(p<0.01)时显著改变了bMECs的形态,以时间依赖性方式增加了细胞凋亡率和MDA含量。在12小时和24小时时,LDH活性和ROS生成也增加(p<0.01)。然而,感染GTII的bMECs中,SOD和CAT含量在12小时时降低(p<0.05),而在24小时时GPx(p<0.01)、SOD(p<0.05)和CAT(p<0.01)水平降低。对于I型基因型(GTI),只有当持续时间延长至24小时时,CAT和GPx活性才显著降低,但低于GTII。这表明GTII对bMECs具有更强的致病作用,本研究结果得出结论,GTII通过氧化还原防御失衡以及细胞内ROS的产生诱导bMECs细胞凋亡和氧化应激。