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Hypoxia in the mammary gland epithelial cells of milk buffalo (BMECs) can affect milk yield and composition, and it can even cause metabolic diseases. Nitidine chloride (NC) is a natural alkaloid with antioxidant properties that can scavenge excessive reactive oxygen species (ROS). However, the effect of NC on the hypoxic injury of BMECs and its molecular mechanisms are still unknown. Here, an immunofluorescence assay, transmission electron microscopy (TEM), and flow cytometry, combined with untargeted metabolomics, were used to investigate the protective effect of NC on hypoxic stress injury in BMECs. It was found that NC can significantly reduce cell activity (p < 0.05) and inhibit cellular oxidative stress (p < 0.05) and cell apoptosis (p < 0.05). A significant decrease in mitophagy mediated by the PINK1-Parkin pathway was observed after NC pretreatment (p < 0.05). In addition, a metabolic pathway enrichment analysis demonstrated that the mechanisms of NC against hypoxic stress may be related to the downregulation of pathways involving aminoacyl tRNA biosynthesis; arginine and proline metabolism; glycine, serine, and threonine metabolism; phenylalanine, tyrosine, and tryptophan biosynthesis; and phenylalanine metabolism. Thus, NC has a protective effect on hypoxic mitochondria, and it can regulate amino acid metabolism in response to hypoxic stress. The present study provides a reference for the application of nitidine chloride to regulate the mammary lactation function of milk buffalo.

Dairy mastitis is an inflammatory reaction caused by mechanical injury and stress within the mammary gland, during which microbial changes and abnormal lipid metabolism occur. However, the underlying mechanism is still unclear. The present study used a combination of 16S rDNA sequencing technology and lipidomics techniques to reveal the effects of mastitis on lactic microbiota and metabolites in the milk of dairy cows. Twenty multiparous Holstein dairy cows (2–3 parities) with an average body weight of 580 ± 30 kg were selected for this study. The dairy cows were allocated to control group (4 cells /mL)) and mastitis group (>5 × 106 cells /mL) based on the somatic cell count. The results showed that mastitis caused a decrease trend in milk production (p = 0.058). The results of the 16 s sequencing indicated a significant decrease (p < 0.05) in the number of Proteobacteria, Tenericutes colonized in mastitis milk, and the number of Firmicutes, Bacteroidetes and Actinobacteria communities increased significantly (p < 0.05). The lipidomics results revealed that the changes in lipid content in mastitis milk were correlated with arachidonic acid metabolism, α -linolenic acid metabolism and glycerol phospholipid metabolism. The results showed that mastitis may cause abnormal lipid metabolism in milk by regulating the diversity of milk microflora, and ultimately affect the milk quality.

Protein nutrition is particularly important for the self-renewal processes of gastrointestinal epithelial cells. The self-renewal of cells is inseparable from the interaction between apoptosis and autophagy. However, there are few reports on the relationship between different nitrogen sources and apoptosis/autophagy. In this study, the relative protein expression of Bcl-2-associated X protein(Bax), caspase-3, and p62 was significantly higher (p < 0.05), while that of Bcl-xl, Bcl-2, Beclin1, and Microtuble-associated protein light chain 3 (LC3-II) was significantly lower (p < 0.05), in the NH4Cl group in comparison with the NH4Cl + 4-phenylbutyric acid (4PBA) group. In addition, the relative protein expression of Bax and caspase-3 was significantly higher (p < 0.05), while that of Bcl-2 and Bcl-xl was decreased significantly (p < 0.05), in the NH4Cl + 3-Methyladenine (3-MA) group and the methionine (Met) + 3-MA group in comparison with the NH4Cl group. Furthermore, the relative protein expression of Beclin1 and LC3B-II was significantly lower (p < 0.05), while that of p62 was significantly higher (p < 0.05), in the NH4Cl + Z-VAD-FMK group and the Met + Z-VAD-FMK group in comparison with the NH4Cl group. In conclusion, our results suggested that endoplasmic reticulum (ER) stress played a critical role in the crosstalk between apoptosis and autophagy induced by NH4Cl and Met. Autophagy had a more obvious ameliorative effect on ruminal epithelial cell apoptosis after treatment with nonprotein nitrogen than after treatment with protein nitrogen. These findings may reveal the molecular mechanism of apoptosis and autophagy induced by nonprotein nitrogen and protein nitrogen.

Changes in the environment such as high-altitude hypoxia (HAH) high-altitude hypoxia can lead to adaptive changes in the blood system of mammals. However, there is limited information about the adaptation of Holstein dairy cows introduced to high-altitude areas. This study used 12 multiparous Holstein dairy cows (600 ± 55 kg, average three years old) exposed to HAH conditions in Nyingchi of Tibet (altitude 3000 m) and HAH-free conditions in Shenyang (altitude 50 m). The miRNA microarray analysis and iTRAQ proteomics approach (accepted as more suitable for accurate and comprehensive prediction of miRNA targets) were applied to explore the differences in the plasma proteomic and miRNA profiles in Holstein dairy cows. A total of 70 differential miRNAs (54 up-regulated, Fold change (FC) FC > 2, and 16 down-regulated, FC < 0.5) and 226 differential proteins (132 up-regulated, FC > 1.2, and 94 down-regulated, FC < 0.8) were found in the HAH-stressed group compared with the HAH-free group. Integrative analysis of proteomic and miRNA profiles demonstrated the biological processes associated with differential proteins were the immune response, complement activation, protein activation, and lipid transport. The integrative analysis of canonical pathways were most prominently associated with the APR signaling (z = 1.604), and LXR/RXR activation (z = 0.365), and FXR/RXR activation (z = 0.446) pathways. The current results indicated that Holstein dairy cows exposed to HAH could adapt to high-altitude hypoxia by up-regulating the APR, activating the LXR/RXR and FXE/RXR pathways.

Nutritional status during mid-gestation is often ignored under extensive husbandry. This study aimed to examine the effect of maternal undernutrition during mid-gestation on kid meat production under an extensive system. Twenty-seven goats (45 ± 3 d of gestation) were randomly assigned to an unrestricted group (100% of nutrient requirements), or a restricted group (60% of nutrient requirements from 45 to 100 d of gestation, and then re-alimented to 100%). Among the offspring, 16 eligible kids (eight per treatment) were selected, based on birth type and survival, and were harvested to evaluate the meat yield, quality, and composition at 90 d after birth. Maternal undernutrition reduced the body weight and size, average daily gain and hot carcass weight of the kids (p < 0.05). The lightness of the meat at 45 min postmortem was increased (p = 0.029) in the restricted kids. Apart from an increase in tyrosine concentration (p = 0.046), the proximate composition and the amino acid and fatty acid profiles were unaffected in the restricted kids (p > 0.05). Overall, maternal undernutrition during mid-gestation decreased the yield of kid meat, but did not significantly modify the quality and composition. These results highlight the importance of nutrient status during mid-gestation in the meat production of small ruminants under an extensive regime.