氨调控宰后牦牛肉中低氧诱导因子-1α表达对线粒体细胞凋亡及嫩度的影响

doi:10.7506/spkx1002-6630-20220710-102

摘要/Abstract

摘要： 为探究氨介导宰后牦牛肉中低氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）表达对线粒体细胞凋亡及肌肉嫩度的影响。以10 mmol/L氯化铵（NH4Cl）、0.9%生理盐水（对照）和10 mmol/L N-硝基-L-精氨酸甲酯盐酸盐（N-nitro-L-arginine methyl ester hydrochloride，L-NAME）3 种溶液处理的牦牛背最长肌为研究对象，通过测定HIF-1α表达量、NO含量、能量代谢酶活力、ATP水平、细胞凋亡线粒体途径及肌肉嫩度等相关指标，探讨宰后牦牛肉细胞凋亡与嫩度变化的机制，完善宰后牦牛肉成熟过程中细胞凋亡与嫩度变化理论体系。结果表明：在宰后牦牛肉成熟过程，各处理组HIF-1α表达量、NO含量总体呈现先增加后减少趋势，Na＋-K＋-ATPase与Ca2＋-ATPase活力、细胞凋亡酶9（Caspase-9）与细胞凋亡酶3（Caspase-3）活力和剪切力呈现先升高后降低的趋势。ATP含量、线粒体膜通透性转换孔（mitochondrial permeability transition pore，MPTP）开放程度、线粒体膜电位（mitochondrial membrane potential，MMP）及肌纤维直径和面积随成熟时间延长呈现逐渐下降趋势，而肌原纤维小片化指数（myofibril fragmentation index，MFI）与肌细胞间隙呈现逐渐增大趋势。宰后6～24 h，NH4Cl组HIF-1α表达量和NO含量显著高于对照组（P＜0.05），L-NAME组显著低于对照组（P＜0.05）。宰后6～72 h，NH4Cl组ATP含量显著高于对照组与L-NAME组。宰后6～24 h，对照组Na＋-K＋-ATPase、Ca2＋-ATPase活力显著高于NH4Cl组（P＜0.05），显著低于L-NAME组（P＜0.05）。宰后6～120 h，NH4Cl组MPTP开放程度显著小于对照组（P＜0.05），L-NAME组显著大于对照组（P＜0.05）。宰后0～168 h，NH4Cl组MMP下降47.72%，对照组下降53.54%，L-NAME组下降60.05%。宰后6～72 h，NH4Cl组Caspase-9与Caspase-3活力显著低于对照组（P＜0.05），L-NAME组Caspase-9与Caspase-3活力显著高于对照组（P＜0.05）；宰后6～120 h，NH4Cl组剪切力、肌纤维面积与直径大于对照组，MFI和肌纤维间隙小于对照组，L-NAME组则相反。综上，随着牦牛肉宰后成熟，氨通过上调HIF-1α表达进而使Na＋-K＋-ATPase与Ca2＋-ATPase活力下降、ATP含量增加，从而调控肌细胞能量代谢，维持内环境稳定；通过抑制MPTP开放以及MMP下降，降低Caspase-3/9活力，抑制线粒体细胞凋亡发生，进而引起剪切力增大、MFI减小以及肌细胞形态变化，最终对牦牛肉嫩度产生负面影响。

关键词: 氨；低氧诱导因子-1α；细胞凋亡；嫩度

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.

Key words: ammonia; hypoxia inducible factor-1α; apoptosis; tenderness

中图分类号:

TS251.5＋2

田凯，辛可启，余群力，韩玲，张新军，孔祥颖，宋仁德. 氨调控宰后牦牛肉中低氧诱导因子-1α表达对线粒体细胞凋亡及嫩度的影响[J]. 食品科学, 2023, 44(15): 1-9.

TIAN Kai, XIN Keqi, YU Qunli, HAN Ling, ZHANG Xinjun, KONG Xiangying, SONG Rende. Ammonia Affects Mitochondrial Cell Apoptosis and Tenderness in Postmortem Yak Meat by Regulating Hypoxia Inducible Factor-1α Expression[J]. FOOD SCIENCE, 2023, 44(15): 1-9.

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氨调控宰后牦牛肉中低氧诱导因子-1α表达对线粒体细胞凋亡及嫩度的影响

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

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