1 College of Life Science and Biotechnology, Gansu Agricultural University, Lanzhou 730070, China; 2 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China; 3 Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou 730070, China
Abstract：Nitric oxide synthase (NOS), a crucial enzyme responsible for production of endogenous nitric oxide (NO) in body, has multiple effects such as anti-inflammatory or pro-inflammatory. To investigate the expression patterns and potential functions of NOS in dairy cows (Bos taurus) with healthy or clinical mastitis (CM), the mammary gland tissues were collected from dairy cows with healthy (control, Con) and CM in this study. The distribution and expression patterns of NOS were in the mammary gland tissues of the Con and CM groups using hematoxylin-eosin (HE) staining, immunohistochemistry (IHC) and real-time fluorescence quantitative PCR (qRT-PCR). The potential role of NOS was predicted using different bioinformatics methods. HE displayed that the mammary acini were collapsed and couple with infiltration inflammatory cell in the CM group. IHC indicated that the positive expression of neuronal NOS (nNOS), inducible NOS (iNOS), and endothelial NOS (eNOS) were presented in the cytoplasm of mammary epithelial cells. Compared to the Con group, the nNOS and eNOS mRNA expression levels were extremely significantly downregulated (P＜0.01), and iNOS mRNA expression levels was extremely significantly upregulated (P＜0.01) in mammary gland tissues of the CM groups. Bioinformatics analysis revealed that NOS might participate in CM occurrence in regulation of NO and amino acid synthesis and metabolism, ion transport. In conclusion, the above results provide a reference for revealing the function and regulatory mechanism of NOS in cow CM.
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