Involvement of Pyocyanin in Promoting LPS-Induced Apoptosis, Inflammation, and Oxidative Stress in Bovine Mammary Epithelium Cells

Author:

Zhu Hao12,Cao Wendi2,Huang Yicai2,Karrow Niel A.3,Yang Zhangping12ORCID

Affiliation:

1. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China

2. Joint International Research Laboratory of Agriculture and Agri-Product Safety, Ministry of Education of China, Yangzhou University, Yangzhou 225009, China

3. Department of Animal Biosciences, University of Guelph, Guelph, ON N1G 2W1, Canada

Abstract

Pyocyanin (PCN) is an extracellular toxin secreted by Pseudomonas aeruginosa (PA), which has redox capacity and disrupts the redox balance of host cells, affecting cell function and leading to cell death. The aim of this experiment was to compare the degree of apoptosis, inflammation, and oxidative stress of bovine mammary epithelium cells (bMECs) induced by lipopolysaccharide (LPS) and pyocyanin (PCN) and to examine whether PCN can promote the apoptosis, inflammation, and oxidative stress of bMECs induced by LPS. In this study, 1 µg/mL LPS and 1 µg/mL PCN were finally selected for subsequent experiments through dose-dependent experiments. In this study, cells were not given any treatment and were used as the control group (NC). The cells were treated with PCN or LPS individually for 6 h as the PCN group (PCN) or the LPS group (LPS), and the combination of LPS and PCN challenge for 6 h as the LPS + PCN (LPS + PCN) group. Compared with the control and LPS groups, PCN resulted in a significantly upregulated expression of genes related to pro-inflammatory (IL-6, TNF-α, MyD88), apoptotic (Bax, Caspase3, Caspase9), as well as protein expression of components in the TLR4/NF-κB signaling pathway (TLR4, p-p65, p65), and p53 signaling pathway (p-p53, p53, Caspase9) (p < 0.05). Moreover, the expression of genes and proteins was significantly upregulated after PCN treatment combined with LPS compared to either LPS or PCN challenge alone (p < 0.05). The stimulation of PCN combined with LPS significantly increased reactive oxygen species (ROS) and malondialdehyde (MDA) production in bovine mammary epithelial cells (bMECs), as well as decreased glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC). Moreover, cells in the LPS + PCN group aggravated oxidative stress and antioxidant inhibition in cells. In addition, the expression of the corresponding genes and proteins related to the Nrf2 pathway (Nrf2, HO-1) was significantly down-regulated in the PCN group as compared to the control group (p < 0.05). Altogether, PCN stimulation exacerbates inflammatory reactions, apoptosis, and oxidative stress reactions, as well as when combined with LPS challenge in bMECs. Therefore, this study indicates that PCN manifests a role in promoting apoptosis, inflammation, and oxidative stress and interacting with LPS to enhance more serious biological stress responses.

Funder

the National Natural Science Foundation of China

Publisher

MDPI AG

Subject

Plant Science,Agronomy and Crop Science,Food Science

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