鲢鱼Cystatin C对冷藏草鱼肉片质构及线粒体途径细胞凋亡的影响

doi:10.7506/spkx1002-6630-20250306-041

食品科学 ›› 2025, Vol. 46 ›› Issue (15): 145-147.doi: 10.7506/spkx1002-6630-20250306-041

鲢鱼Cystatin C对冷藏草鱼肉片质构及线粒体途径细胞凋亡的影响

李龙飞，李树红，钱邓帆，蒋宇维，仝浩楠，李晓菲，白水萌，张颖，李冉

（四川农业大学食品学院，四川雅安 625000）

出版日期:2025-08-15 发布日期:2025-07-22
基金资助:
四川省自然科学基金面上项目（2025ZNSFSC0221）；四川省自然科学基金青年科学基金项目（2023NSFSC1160）

Effect of Silver Carp Cystatin C on the Texture and Mitochondrial Pathway-Mediated Apoptosis of Refrigerated Grass Carp Fillets

LI Longfei, LI Shuhong, QIAN Dengfan, JIANG Yuwei, TONG Haonan, LI Xiaofei, BAI Shuimeng, ZHANG Ying, LI Ran

(College of Food Science, Sichuan Agricultural University, Ya’an 625000, China)

Online:2025-08-15 Published:2025-07-22

摘要/Abstract

摘要： 分析鲢鱼Cystatin C（HmCystatin C）对冷藏鱼肉片质构品质的保持效果，及其对基于线粒体途径的肌肉细胞凋亡的调节作用，旨在探究HmCystatin C作为冷藏水产品潜在天然生物保鲜剂的可能作用机制。实验首先对草鱼肉片进行HmCystatin C注射处理，在经Western blot验证HmCystatin C细胞渗透有效性的基础上，从质构、组织学特性、组织蛋白酶B/L酶活性及活性氧（reactive oxygen species，ROS）水平监测4 ℃冷藏72 h内鱼肉片品质的变化。进而通过免疫组化法、Tunel染色法、分光光度法及透射电镜检测冷藏期间肌肉细胞凋亡、线粒体膜通透性转换孔（mitochondrial permeability transition pore，MPTP）开放性、线粒体膜电位以及线粒体形态损伤程度，同时利用荧光测活法和Western blot分析Caspase-3、Caspase-9活性及蛋白表达量变化。结果表明HmCystatin C能有效渗透至组织细胞内并持续存在，实验组（Cys组）冷藏期间的剪切力和硬度均显著高于对照（Con组），而细胞间隙、组织蛋白酶B/L活性及ROS水平均明显低于Con组，尤其质构劣化被延缓至少24 h；且HmCystatin C明显抑制了Cys组的线粒体功能和形态的损伤程度。同时Cys组的Caspase-3、Caspase-9活性被不同程度地抑制，蛋白表达量亦低于Con组。综上推测抑制线粒体途径细胞凋亡及其相关Caspases活性是HmCystatin C延缓冷藏草鱼肉片品质劣化的机制之一。

关键词: 鲢鱼Cystatin C；冷藏草鱼肉片；质构特性；线粒体途径细胞凋亡；细胞凋亡酶

Abstract: With the aim of understanding the mechanism of action of cystatin C of silver carp (HmCystatin C) as a potential natural biological preservative, we investigated the effect of HmCystatin C on maintaining the texture quality of refrigerated fish fillets and the regulatory effect on myocyte apoptosis mediated by mitochondrial pathway. Western blot was used to verify the cellular permeability of HmCystatin C in grass carp fillets. Quality changes during refrigeration for 72 h at 4 ℃ were monitored with respect to texture, histological characteristics, cathepsin B/L activity and reactive oxygen species (ROS) levels. Myocyte apoptosis, the opening degree of mitochondrial permeability transition pore (MPTP), mitochondrial membrane potential and mitochondrial morphological damage were detected by immunohistochemistry, TUNEL staining, spectrophotometry, and transmission electron microscopy (TEM). Meanwhile, the activities and protein expression of caspase-3 and caspase-9 were analyzed by fluorescence assay and Western blot, respectively. The results demonstrated that HmCystatin C effectively penetrated into and persistently existed in cells. Compared with the control group, shear force and hardness obviously increased in the Cys treated group, gaps between myocytes, cathepsin B/L activity and ROS levels decreased, and texture deterioration was postponed by at least 24 h. Additionally, the damage degree of mitochondrial function and morphology was suppressed significantly in the Cys group. Meanwhile, the activities of caspase-3 and caspase-9 were inhibited to different extents and the protein expression levels were lower than that in the control group. In conclusion, the inhibition of mitochondrial pathway-mediated apoptosis and relevant caspases activities by HmCystatin C might contribute to its effect on delaying the quality deterioration of refrigerated grass carp fillets.

Key words: cystatin C of silver carp; refrigerated grass carp fillets; texture characteristics; mitochondrial pathway-mediated apoptosis; caspases

中图分类号:

S983

李龙飞，李树红，钱邓帆，蒋宇维，仝浩楠，李晓菲，白水萌，张颖，李冉. 鲢鱼Cystatin C对冷藏草鱼肉片质构及线粒体途径细胞凋亡的影响[J]. 食品科学, 2025, 46(15): 145-147.

LI Longfei, LI Shuhong, QIAN Dengfan, JIANG Yuwei, TONG Haonan, LI Xiaofei, BAI Shuimeng, ZHANG Ying, LI Ran. Effect of Silver Carp Cystatin C on the Texture and Mitochondrial Pathway-Mediated Apoptosis of Refrigerated Grass Carp Fillets[J]. FOOD SCIENCE, 2025, 46(15): 145-147.

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鲢鱼Cystatin C对冷藏草鱼肉片质构及线粒体途径细胞凋亡的影响

Effect of Silver Carp Cystatin C on the Texture and Mitochondrial Pathway-Mediated Apoptosis of Refrigerated Grass Carp Fillets

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