冷鲜牛肉贮藏过程中线粒体变化与肌红蛋白氧化状态的相关性
|
摘要
为探索冷鲜牛肉贮藏过程中线粒体功能结构的变化,采用透射电子显微镜与原子力显微镜对线粒体内、外部超微结构的检测来确定贮藏时间对肌细胞线粒体变化的影响。结果发现:在4℃贮藏过程中(0～21 d),Flameng线粒体评分从0.986 8升高到2.533 8,表明线粒体结构完整性逐渐降低;线粒体膜电位与线粒体呼吸控制率、ATP含量均呈极显著正相关,相关系数分别达到0.936、0.926(P<0.01),说明线粒体结构与功能之间关系密切;线粒体呼吸控制率与氧合肌红蛋白相对含量呈极显著正相关(r=0.885)(P<0.01),线粒体膜电位与高铁肌红蛋白相对含量呈极显著负相关(r=-0.902)(P<0.01),表明线粒体结构、功能变化与肉色稳定性有密切的关系。
In order to explore changes in mitochondrial function and structure during the chilled storage of beef, the mitochondrial damage was evaluated by observation of the internal and external ultrastructure of mitochondria using transmission electron microscopy and atomic force microscopy. With the increase of storage time up to 21 d, Flameng score was increased from 0.986 8 to 2.533 8, indicating that the mitochondrial structural integrity gradually decreased; there were significant positive correlations between mitochondrial membrane potential and mitochondrial respiratory control ratio and ATP content with correlation coefficients of up to 0.936 and 0.926(P < 0.01), reflecting the close relationship between mitochondrial structure and function. Moreover, mitochondrial respiratory control ratio was significantly positively correlated with the relative content oxymyoglobin(r = 0.885, P < 0.01), while mitochondrial membrane potential was significantly negatively correlated with the relative content of metmyoglobin(r = 0.902, P < 0.01), indicating that changes in mitochondrial structure and function are closely related to meat color stability.
In order to explore changes in mitochondrial function and structure during the chilled storage of beef, the mitochondrial damage was evaluated by observation of the internal and external ultrastructure of mitochondria using transmission electron microscopy and atomic force microscopy. With the increase of storage time up to 21 d, Flameng score was increased from 0.986 8 to 2.533 8, indicating that the mitochondrial structural integrity gradually decreased; there were significant positive correlations between mitochondrial membrane potential and mitochondrial respiratory control ratio and ATP content with correlation coefficients of up to 0.936 and 0.926(P < 0.01), reflecting the close relationship between mitochondrial structure and function. Moreover, mitochondrial respiratory control ratio was significantly positively correlated with the relative content oxymyoglobin(r = 0.885, P < 0.01), while mitochondrial membrane potential was significantly negatively correlated with the relative content of metmyoglobin(r = 0.902, P < 0.01), indicating that changes in mitochondrial structure and function are closely related to meat color stability.
引文
[1]MCKEITH R O,KING D A,GRAYSON A L,et al.Influence of mitochondrial efficiency on beef lean color stability[J].Meat Science,2014,96(1):476.DOI:10.1016/j.meatsci.2013.07.116.
[2]BELSKIE K M,VAN BUITEN C B,RAMANATHAN R,et al.Reverse electron transport effects on NADH formation and metmyoglobin reduction[J].Meat Science,2015,105:89-92.DOI:10.1016/j.meatsci.2015.02.012.
[3]RAMANATHAN R,MANCINI R A,KONDA M R.Effects of lactate on beef heart mitochondrial oxygen consumption and muscle darkening[J].Journal of Agricultural and Food Chemistry,2009,57(4):1550-1555.DOI:10.1021/jf802933p.
[4]GAO X G,XIE L,WANG Z Y,et al.Effect of postmortem time on the metmyoglobin reductase activity,oxygen consumption,and colour stability of different lamb muscles[J].European Food Research and Technology,2013,236(4):579-587.DOI:10.1007/s00217-012-1903-8.
[5]MANCINI R A,RAMANATHAN R.Effects of postmortem storage time on color and mitochondria in beef[J].Meat Science,2014,98(1):65-70.DOI:10.1016/j.meatsci.2014.04.007.
[6]WITTENBERG J B,WITTENBERG B A.Myoglobin-enhanced oxygen delivery to isolated cardiac mitochondria[J].Journal of Experimental Biology,2007,210(12):2082-2090.DOI:10.1242/jeb.003947.
[7]TANG J,FAUSTMAN C,HOAGLAND T A,et al.Postmortem oxygen consumption by mitochondria and its effects on myoglobin form and stability[J].Journal of Agricultural and Food Chemistry,2005,53(4):1223-1230.DOI:10.1021/jf048646o.
[8]TANG J,FAUSTMAN C,MANCINI R A,et al.Mitochondrial reduction of metmyoglobin:dependence on the electron transport chain[J].Journal of Agricultural and Food Chemistry,2005,53(13):5449-5455.DOI:10.1021/jf050092h.
[9]展平.姜黄素激活Nrf2系统缓解线粒体损伤和胰岛素抵抗作用机制研究[D].福州:福建中医药大学,2017:21-23.
[10]赵静,万绍贵,鲍登克.线粒体损伤相关模式分子与宿主免疫调节[J].中国生物化学与分子生物学报,2017,33(1):38-43.DOI:10.13865/j.cnki.cjbmb.2017.01.06.
[11]刘红杰,詹莎,李天昊,等.艾叶挥发油致急性肝损伤小鼠线粒体结构和功能的变化[J].中国临床药理学杂志,2017,33(6):530-534.DOI:10.13699/j.cnki.1001-6821.2017.06.014.
[12]蒋康伦.快速衰老小鼠张口呼吸模型制备及其线粒体功能损伤研究[D].镇江:江苏大学,2017:15-19.
[13]劳文艳,毕婷婷,周艳丽,等.阿魏酸拮抗PM2.5对A549细胞线粒体的损伤作用[J].食品科学,2017,38(3):195-200.DOI:10.7506/spkx1002-6630-201703032.
[14]曹锦轩,张玉林,黄鸿兵,等.宰后骨骼肌内环境对caspase-3活化的潜在影响因素综述[J].食品与发酵工业,2013,39(1):155-160.DOI:10.13995/j.cnki.11-1802/ts.2013.01.047.
[15]王静,王玲,王自彬.线粒体自噬过程的超微结构分析[J].中国细胞生物学学报,2017,39(4):435-439.DOI:11844/cjcb.2017.04.0318.
[16]史华伟,郭蓉娟,赵振武,等.醒脾解郁方对抑郁大鼠行为学及海马与骨骼肌线粒体超微结构的影响[J].北京中医药大学学报,2017,40(4):284-289.
[17]宋冬梅,陈丽,马俊香,等.锰对SH-SY5Y细胞线粒体超微结构及功能改变的研究[J].实用医学杂志,2017,33(5):680-683.DOI:10.3969/j.issn.1006?5725.2017.05.002.
[18]朱龙飞,田军,坚哲,等.黄芩素对氧化应激状态下白癜风黑素细胞线粒体超微结构和功能的影响[J].中国麻风皮肤病杂志,2017,33(6):321-324.
[19]王梓,杨欣莹,吕楠,等.血府逐瘀胶囊对大鼠急性心肌缺血及心肌线粒体超微结构的影响[J].中国中医基础医学杂志,2017,23(8):1067-1070.
[20]陈松,韩永丽,吴松,等.电针内关穴预处理对大鼠心肌缺血再灌注后线粒体功能及超微结构的保护作用[J].中华中医药杂志,2017,32(10):4651-4654.
[21]MCKEITH R O,KING D A,GRAYSON A L,et al.Mitochondrial abundance and efficiency contribute to lean color of dark cutting beef[J].Meat Science,2016,116:165-173.DOI:10.1016/j.meatsci.2016.01.016.
[22]GAGAOUA M,TERLOUW E M,MICOL D,et al.Understanding early post-mortem biochemical processes underlying Meat color and pH decline in the Longissimus thoracis muscle of young blond d’Aquitaine bulls using protein biomarkers[J].Journal of Agricultural and Food Chemistry,2015,63(30):6799-6809.DOI:10.1021/acs.jafc.5b02615.
[23]WYRWISZ J,MOCZKOWSKA M,KUREK M,et al.Influence of21 days of vacuum-aging on color,bloom development,and WBSFof beef semimembranosus[J].Meat Science,2016,122:48-54.DOI:10.1016/j.meatsci.2016.07.018.
[24]KRZYWICKI K.The determination of haem pigments in meat[J].Meat Science,1982,7(1):29-36.
[25]WITTENBERG J B,WITTENBERG B A.Myoglobin-enhanced oxygen delivery to isolated cardiac mitochondria[J].Journal of Experimental Biology,2007,210(12):2082-2090.DOI:10.1242/jeb.003947.
[26]彭艳艳,孙丽娜,宋菁,等.低氧对小鼠骨骼肌离体线粒体膜通透性转换孔功能的影响[J].温州医学院学报,2012,42(2):110-113.DOI:10.3969/j.issn.1000-2138.2012.02.003.
[27]赵丽红.食源肥胖鼠卵母细胞线粒体相关内质网膜结构功能的研究[D].北京:中国农业大学,2017:21-26.
[28]李兴泰,冮洁,匡海学,等.红豆越橘果总黄酮保护线粒体及其机制[J].食品科学,2013,34(19):67-72.DOI:10.7506/spkx1002-6630-201319015.
[29]凌曦.PAHs致男(雄)性生殖损害中端粒和线粒体损伤效应和机制研究[D].重庆:第三军医大学,2017:18-26.
[30]吴琼.参知健脑方对缺氧致认知功能障碍大鼠线粒体损伤的保护机制的实验研究[D].北京:北京中医药大学,2017:17-20.
[31]田金强,朱克瑞,李新明,等.两种多糖对γ辐射诱导的鼠肝线粒体伤害的抑制[J].食品科学,2006,27(5):235-239.
[2]BELSKIE K M,VAN BUITEN C B,RAMANATHAN R,et al.Reverse electron transport effects on NADH formation and metmyoglobin reduction[J].Meat Science,2015,105:89-92.DOI:10.1016/j.meatsci.2015.02.012.
[3]RAMANATHAN R,MANCINI R A,KONDA M R.Effects of lactate on beef heart mitochondrial oxygen consumption and muscle darkening[J].Journal of Agricultural and Food Chemistry,2009,57(4):1550-1555.DOI:10.1021/jf802933p.
[4]GAO X G,XIE L,WANG Z Y,et al.Effect of postmortem time on the metmyoglobin reductase activity,oxygen consumption,and colour stability of different lamb muscles[J].European Food Research and Technology,2013,236(4):579-587.DOI:10.1007/s00217-012-1903-8.
[5]MANCINI R A,RAMANATHAN R.Effects of postmortem storage time on color and mitochondria in beef[J].Meat Science,2014,98(1):65-70.DOI:10.1016/j.meatsci.2014.04.007.
[6]WITTENBERG J B,WITTENBERG B A.Myoglobin-enhanced oxygen delivery to isolated cardiac mitochondria[J].Journal of Experimental Biology,2007,210(12):2082-2090.DOI:10.1242/jeb.003947.
[7]TANG J,FAUSTMAN C,HOAGLAND T A,et al.Postmortem oxygen consumption by mitochondria and its effects on myoglobin form and stability[J].Journal of Agricultural and Food Chemistry,2005,53(4):1223-1230.DOI:10.1021/jf048646o.
[8]TANG J,FAUSTMAN C,MANCINI R A,et al.Mitochondrial reduction of metmyoglobin:dependence on the electron transport chain[J].Journal of Agricultural and Food Chemistry,2005,53(13):5449-5455.DOI:10.1021/jf050092h.
[9]展平.姜黄素激活Nrf2系统缓解线粒体损伤和胰岛素抵抗作用机制研究[D].福州:福建中医药大学,2017:21-23.
[10]赵静,万绍贵,鲍登克.线粒体损伤相关模式分子与宿主免疫调节[J].中国生物化学与分子生物学报,2017,33(1):38-43.DOI:10.13865/j.cnki.cjbmb.2017.01.06.
[11]刘红杰,詹莎,李天昊,等.艾叶挥发油致急性肝损伤小鼠线粒体结构和功能的变化[J].中国临床药理学杂志,2017,33(6):530-534.DOI:10.13699/j.cnki.1001-6821.2017.06.014.
[12]蒋康伦.快速衰老小鼠张口呼吸模型制备及其线粒体功能损伤研究[D].镇江:江苏大学,2017:15-19.
[13]劳文艳,毕婷婷,周艳丽,等.阿魏酸拮抗PM2.5对A549细胞线粒体的损伤作用[J].食品科学,2017,38(3):195-200.DOI:10.7506/spkx1002-6630-201703032.
[14]曹锦轩,张玉林,黄鸿兵,等.宰后骨骼肌内环境对caspase-3活化的潜在影响因素综述[J].食品与发酵工业,2013,39(1):155-160.DOI:10.13995/j.cnki.11-1802/ts.2013.01.047.
[15]王静,王玲,王自彬.线粒体自噬过程的超微结构分析[J].中国细胞生物学学报,2017,39(4):435-439.DOI:11844/cjcb.2017.04.0318.
[16]史华伟,郭蓉娟,赵振武,等.醒脾解郁方对抑郁大鼠行为学及海马与骨骼肌线粒体超微结构的影响[J].北京中医药大学学报,2017,40(4):284-289.
[17]宋冬梅,陈丽,马俊香,等.锰对SH-SY5Y细胞线粒体超微结构及功能改变的研究[J].实用医学杂志,2017,33(5):680-683.DOI:10.3969/j.issn.1006?5725.2017.05.002.
[18]朱龙飞,田军,坚哲,等.黄芩素对氧化应激状态下白癜风黑素细胞线粒体超微结构和功能的影响[J].中国麻风皮肤病杂志,2017,33(6):321-324.
[19]王梓,杨欣莹,吕楠,等.血府逐瘀胶囊对大鼠急性心肌缺血及心肌线粒体超微结构的影响[J].中国中医基础医学杂志,2017,23(8):1067-1070.
[20]陈松,韩永丽,吴松,等.电针内关穴预处理对大鼠心肌缺血再灌注后线粒体功能及超微结构的保护作用[J].中华中医药杂志,2017,32(10):4651-4654.
[21]MCKEITH R O,KING D A,GRAYSON A L,et al.Mitochondrial abundance and efficiency contribute to lean color of dark cutting beef[J].Meat Science,2016,116:165-173.DOI:10.1016/j.meatsci.2016.01.016.
[22]GAGAOUA M,TERLOUW E M,MICOL D,et al.Understanding early post-mortem biochemical processes underlying Meat color and pH decline in the Longissimus thoracis muscle of young blond d’Aquitaine bulls using protein biomarkers[J].Journal of Agricultural and Food Chemistry,2015,63(30):6799-6809.DOI:10.1021/acs.jafc.5b02615.
[23]WYRWISZ J,MOCZKOWSKA M,KUREK M,et al.Influence of21 days of vacuum-aging on color,bloom development,and WBSFof beef semimembranosus[J].Meat Science,2016,122:48-54.DOI:10.1016/j.meatsci.2016.07.018.
[24]KRZYWICKI K.The determination of haem pigments in meat[J].Meat Science,1982,7(1):29-36.
[25]WITTENBERG J B,WITTENBERG B A.Myoglobin-enhanced oxygen delivery to isolated cardiac mitochondria[J].Journal of Experimental Biology,2007,210(12):2082-2090.DOI:10.1242/jeb.003947.
[26]彭艳艳,孙丽娜,宋菁,等.低氧对小鼠骨骼肌离体线粒体膜通透性转换孔功能的影响[J].温州医学院学报,2012,42(2):110-113.DOI:10.3969/j.issn.1000-2138.2012.02.003.
[27]赵丽红.食源肥胖鼠卵母细胞线粒体相关内质网膜结构功能的研究[D].北京:中国农业大学,2017:21-26.
[28]李兴泰,冮洁,匡海学,等.红豆越橘果总黄酮保护线粒体及其机制[J].食品科学,2013,34(19):67-72.DOI:10.7506/spkx1002-6630-201319015.
[29]凌曦.PAHs致男(雄)性生殖损害中端粒和线粒体损伤效应和机制研究[D].重庆:第三军医大学,2017:18-26.
[30]吴琼.参知健脑方对缺氧致认知功能障碍大鼠线粒体损伤的保护机制的实验研究[D].北京:北京中医药大学,2017:17-20.
[31]田金强,朱克瑞,李新明,等.两种多糖对γ辐射诱导的鼠肝线粒体伤害的抑制[J].食品科学,2006,27(5):235-239.