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| The Regulatory Role of RAS in LPS-induced Inflammatory of bMECs |
| ZHANG Xiang-Jun, ZHOU Xue-Zhang* |
| College of Life Science, Ningxia University / Key Laboratory of the Ministry of Education for the Conservation and Utilization of Special Biological Resources of Western China, Yinchuan 750021, China |
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Abstract The aim of this study is to explore the renin-angiotensin system's (RAS) expression and location in bovine mammary epithelial cells (bMECs), and its regulatory role in LPS-induced inflammatory injury. After treating bMECs with different concentrations of LPS, CCK-8 method was used to examine cell viability, and qPCR was used to detect IL-1β, IL-6, IL-8, TNF-α, ACE2, ACE, MasR和AT1R mRNA expression. Enzyme linked immunosorbent assay (ELISA) was used to test the content of AngⅡ and Ang 1-7 protein in the supernatant. Western blot and cellular immunofluorescence were used to verify the expression and location of ACE2 protein in bMECs. The results showed that ACE2 was expressed and localized in the cell membrane and cytoplasm of bMECs. Producing obvious inflammatory damage by LPS treatment of bMECs for 24 h, the expression of IL-1β, IL-6, IL-8, TNF-α, ACE, AT1R gene, and AngⅡ proein increased with the LPS concentration. The expression of ACE2 and MasR mRNA firstly increased and then decreased, while the expression of Ang 1-7 protein decreased along with the LPS concentration. This study indicates the presence of RAS members such as ACE2, ACE, MasR, AT1R, AngⅡ and Ang 1-7 in bMECs, and the ACE2/MasR/Ang 1-7 and ACE/Ang Ⅱ/AT1R pathways involved in the regulation of bMECs inflammatory injury induced by LPS. This experiment lays the foundation for studying the regulatory role of RAS in mastitis and discovering new targets for the action of anti-inflammatory drug.
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Received: 07 January 2021
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Corresponding Authors:
* zhouxuezhang@nxu.edu.cn
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