No abstract

Ammonia, produced mainly from the deamination of amino acids and glutamine, is one of the major toxic components in blood and tissues that may affect bovine health. However, the physiological and pathological roles of ammonia in the mammary glands are not understood clearly. In the present study, the bovine mammary epithelial cell line (MAC-T) was utilised as an in vitro model to determine the effects of ammonia on bovine mammary gland. We demonstrated that ammonia stimulated the production of intracellular reactive oxygen species, decreased mitochondrial membrane potential, interrupted intracellular calcium ion (Ca2+) homeostasis and induced cell apoptosis. Ammonia also significantly reduced cell viability and increased the proportion of apoptotic cells through enhancing the level of p53 phosphorylation and increasing the expressions of BAX, caspase 8, caspase 9, caspase 3. Interestingly, bumetanide, a specific Na+ K+ 2Cl--cotransporter inhibitor, dramatically abolished the damaging effects of ammonia on the cells. These data suggest that ammonia exposure induces apoptosis in bovine mammary epithelial cells via activation of the p53 pathway and the mitochondrial apoptotic pathway, and that these effects involved the Na+ K+ 2Cl--cotransporter.

The present study was designed to explore the therapeutic potential of self-assembling peptide nanofiber scaffold (SAPNS) delivered RhoA inhibitor to ameliorate the hostile microenvironment of injured spinal cord for axonal regeneration. After a transection was applied to the thoracic spinal cord of mice, the combination of SAPNS and CT04 (a cell permeable RhoA inhibitor), single SAPNS with vehicle, or saline was transplanted into the lesion cavity. Results showed that SAPNS+CT04 implants achieved the best therapeutic outcomes among treatment groups. The novel combination not only reconstructed the injured nerve gap but also elicited significant axonal regeneration and motor functional recovery. Additionally, the combination also effectively reduced the apoptosis and infiltration of activated macrophages in the injured spinal cord. Collectively, the present study demonstrated that SAPNS-based delivery of RhoA inhibitor CT04 presented a highly potential therapeutic strategy for spinal cord injury with reknitting lesion gap, attenuating secondary injury, and improving axonal regrowth.