Ammonia Affects Mitochondrial Cell Apoptosis and Tenderness in Postmortem Yak Meat by Regulating Hypoxia Inducible Factor-1α Expression

Abstract

This study aimed to investigate the effect of ammonia mediated hypoxia inducible factor-1α (HIF-1α) expression on mitochondrial cell apoptosis and tenderness in yak meat after slaughter. Ten mmol/L ammonium chloride (NH4Cl), 0.9% normal saline (control) and 10 mmol/L N-nitro-L-arginine methyl ester hydrochloride were used separately for treating yak Longissimus dorsi. By measuring the expression of HIF-1α, nitrogen monoxide (NO) content, energy metabolism-related enzyme activities, adenosine triphosphate (ATP) level, mitochondrial pathway of apoptosis and muscle tenderness, we explored the mechanism of cell apoptosis and tenderness change in yak meat after slaughter. The results showed that during the postmortem aging of yak meat, the expression of HIF-1α and the content of NO in each treatment group increased first and then decreased, and so did the activities of Na+-K+-ATPase, Ca2+-ATPase, caspase-9 and caspase-3 and shear stress. ATP content, the degree of mitochondrial permeability transition pore (MPTP) opening, mitochondrial membrane potential (MMP), and muscle fiber diameter and area showed a gradual downward trend, and myofibril fragmentation index (MFI) and the gap between muscle cells showed an opposite trend. During 6 to 24 h after slaughter, the expression of HIF-1α and the content of NO in the NH4Cl treatment group were significantly higher than those in the control group (P < 0.05), which were significantly higher than those in the L-NAME treatment group (P < 0.05). The ATP content in the NH4Cl group was significantly higher than that in the control and L-NAME groups during 6 to 72 h after slaughter. The activities of Na+-K+-ATPase and Ca2+-ATPase in the control group were significantly higher than those in the NH4Cl group (P < 0.05), but significantly lower than those in the L-NAME group (P < 0.05). During 6–120 h after slaughter, the opening of MPTP in the control group was significantly higher than that in the NH4Cl group (P < 0.05), but lower than that in the L-NAME group (P < 0.05). During 0 to 168 h after slaughter, the opening of MMP decreased by 47.72% in the NH4Cl group, 53.54% in the control group, and 60.05% in the L-NAME group. During 6 to 72 h after slaughter, the activities of caspase-9 and caspase-3 in the NH4Cl group were significantly lower than those in the control group (P < 0.05), which were lower than those in the L-NAME group (P < 0.05). During 6 to 120 h after slaughter, shear force and muscle fiber area and diameter in the NH4Cl group were higher than those in the control group, and MFI and the gap between muscle fibers in the NH4Cl group were lower than those in the control group, while the opposite result was observed for the L-NAME group. In conclusion, ammonia decreased the activities of Na+-K+-ATPase and Ca2+-ATPase and increased the content of ATP by up-regulating the expression of HIF-1α, which in turn regulated the energy metabolism of muscle cells and maintained the stability of the internal environment. By inhibiting the opening of MPTP and the decrease in MMP, ammonia reduced the activity of caspase-3/9, and inhibited mitochondrial cell apoptosis, thereby resulting in an increase in shear stress, a decrease in MFI and morphological changes of muscle cells, and finally causing a negative effect on the tenderness of yak meat.

Keywords

• ammonia; hypoxia inducible factor-1α; apoptosis; tenderness