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溶酶體組織蛋白酶參與宰后牛肉AIF介導(dǎo)細(xì)胞凋亡研究
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國(guó)家自然科學(xué)基金項(xiàng)目(31560463、31460402)和國(guó)家肉牛牦牛產(chǎn)業(yè)技術(shù)體系項(xiàng)目(CARS-38)


Lysosomal Cathepsins in Apoptosis-inducing-factor-mediate Apoptosis of Bovine Muscle during Postmortem Aging
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    摘要:

    為研究牛肉宰后成熟過程中凋亡誘導(dǎo)因子(AIF)介導(dǎo)細(xì)胞凋亡及溶酶體組織蛋白酶參與AIF活化機(jī)制,,選取6頭西門塔爾雜交牛進(jìn)行屠宰,,取其背最長(zhǎng)肌于4℃條件下成熟,,在不同成熟時(shí)間點(diǎn)(0,、6,、12,、24,、72,、120,、168h)進(jìn)行活性氧(ROS)相對(duì)含量、溶酶體膜穩(wěn)定性,、組織蛋白酶活性,、線粒體膜通透性、AIF表達(dá)及細(xì)胞核形態(tài)的測(cè)定,。結(jié)果表明,,宰后成熟過程中,牛肉ROS相對(duì)含量呈先降低后升高的趨勢(shì)(P<0.05),;溶酶體膜穩(wěn)定性逐漸降低(P<0.05),;組織蛋白酶B、D和L的活性逐漸升高(P<0.05),;線粒體膜通透性增大(P<0.05),;AIF在線粒體中的表達(dá)量于宰后12h顯著升高,在之后的成熟時(shí)間降低(P<0.05),AIF在細(xì)胞核中的表達(dá)呈上升的趨勢(shì)(P<0.05),,與線粒體中的下降趨勢(shì)相一致,;細(xì)胞核體積逐漸增大、部分核溶解,,并伴隨細(xì)胞間隙變大的現(xiàn)象,。表明牛肉在宰后成熟過程中生成ROS,ROS的累積導(dǎo)致溶酶體膜穩(wěn)定性失衡,,使組織蛋白酶被釋放,,作用于線粒體,之后線粒體膜通透性發(fā)生變化,,釋放AIF,,活化的AIF進(jìn)入細(xì)胞核內(nèi)誘導(dǎo)凋亡發(fā)生。

    Abstract:

    With the aim to investigate apoptosis-inducing-factor (AIF)-mediate apoptosis and mechanism of lysosomal cathepsins involved in AIF activation of bovine muscle during postmortem aging, six healthy Simmental crossbreeding local yellow cattle were slaughtered and longissimus thoracic muscles were removed from the carcasses then aged at 4℃ for 6h, 12h, 24h, 72h, 120h and 168h. At the end of each storage period, ROS content, lysosomal membrane stability, cathepsins activities, mitochondrial membrane permeability, the expression of AIF and nuclear morphology were measured. The results suggested that ROS content was decreased from 0h to 6h, but it was significantly increased from 6h to 168h during postmortem aging (P<0.05). Lysosomal membrane stability was significantly decreased from 0h to 168h (P<0.05). Cathepsin B, D and L activities were increased as the postmortem aging was increased (P<0.05). Mitochondrial membrane permeability was significantly increased during postmortem aging (P<0.05). The AIF level in mitochondria was significantly increased from 0h to 12h and decreased as the postmortem aging was increased, the amount of AIF in the nucleus was increased during postmortem aging (P<0.05). The volume of cell nucleus was gradually increased and some nucleus were dissolved and broken during bovine postmortem aging. It was concluded that the generation of ROS may lead to imbalance of lysosomal membrane stability, and then cathepsins were released to cytoplasm. Subsequently, the mitochondrial outer membrane was permeabilized, and AIF was translocated to the cytosol and nucleus, where it induced apoptosis of nucleus.

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張佳瑩,余群力,韓玲,李航,殷元虎,韓明山.溶酶體組織蛋白酶參與宰后牛肉AIF介導(dǎo)細(xì)胞凋亡研究[J].農(nóng)業(yè)機(jī)械學(xué)報(bào),2018,49(3):354-360. ZHANG Jiaying, YU Qunli, HAN Ling, LI Hang, YIN Yuanhu, HAN Mingshan. Lysosomal Cathepsins in Apoptosis-inducing-factor-mediate Apoptosis of Bovine Muscle during Postmortem Aging[J]. Transactions of the Chinese Society for Agricultural Machinery,2018,49(3):354-360.

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  • 收稿日期:2017-07-30
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  • 在線發(fā)布日期: 2018-03-10
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