藏羊肉宰后成熟过程中HSP27对肌原纤维蛋白及细胞凋亡酶的影响

摘要/Abstract

摘要： 为研究藏羊肉宰后成熟过程中热休克蛋白27（heat shock protein 27，Hsp27）对肌原纤维蛋白降解特性及细胞凋亡酶活性的影响。以经Hsp27的抑制剂KRIBB3处理的藏羊肉为研究对象，测定藏羊肉在宰后成熟过程中Caspase-3、9的活性变化以及肌原纤维蛋白的溶解性和降解特性，并通过体外试验进一步研究Hsp27对μ-钙激活酶和Caspase-3降解肌原纤维蛋白的影响情况。结果表明，Hsp27能显著抑制Caspase-3、9的活性（p < 0.05），能够抑制肌原纤维小片化的发生以及肌间线蛋白和肌钙蛋白-T的降解；体外试验研究发现，Hsp27能够抑制μ-钙激活酶和Caspase-3对肌原纤维蛋白的降解。

关键词: 藏羊肉, 热休克蛋白27, 细胞凋亡酶, 免疫印迹（WB）, 肌原纤维

Abstract: In order to study the effects of heat shock protein 27 (Hsp27) on the degradation of myofibrillar proteins and the activity of apoptotic enzymes during the postmortem maturation of Tibetan mutton. The Tibetan mutton treated with Hsp27 inhibitor KRIBB3 was used as the research object to determine the activity changes of Caspase-3 and 9 and the solubility and degradation characteristics of myofibrillar protein during postmortem maturation of Tibetan mutton, and further study by in vitro test. Effect of Hsp27 on the degradation of myofibrillar protein by μ-calcium activase and Caspase-3. The results showed that Hsp27 significantly inhibited the activity of Caspase-3 and 9 (p < 0.05), inhibited the formation of myofibril fragmentation and the degradation of myologin and troponin-T. In vitro studies found that Hsp27 was able to Significant inhibition of μ-calcium activase and Caspase-3 degradation of myofibrillar proteins.

Key words: Tibetan sheep meat, Heat shock protein 27 (Hsp27), Caspases, Western blotting, Myofibrillar

中图分类号:

孙金龙师希雄黄峰韩玲岳建伟陈骋. 藏羊肉宰后成熟过程中HSP27对肌原纤维蛋白及细胞凋亡酶的影响[J]. 食品科学, 0, (): 0-0.

参考文献

[1]闫忠心, 靳义超.基于氨基酸和脂肪酸的藏羊肉质量评价[J]. [J].食品工业科技, 2016, 37((3)):351-354 [2]皮立, 胡凤祖, 星玉秀, 等.青海欧拉羊肌肉脂肪酸组成的气相色谱-质谱分析[J]. [J].食品科学, 2012, 33((6)): 190-193 [3]王振宇, 王培培, 陈丽, 等.欧拉型藏羊肉理化及近红外光谱特性研究[J].[J].2012, 肉类研究, 26((9)):1-4 [4]李胜杰, 胡忠良, 郭向莹, 等.细胞凋亡对肌肉宰后嫩化的贡献[J]. 食品工业科技, 2010(4):395-398. [5]KOOHMARRAIE, M & GEESINK, G H (2006).Contribution of postmortem muscle biochemistry to the delivery of consistent meat quality with particular focus on the calpain system. Meat Science, 74, 34–43. [6]COSTELLI P, REFFO P, PENNA F, et al.Ca2+-dependent proteolysis in muscle wasting[J].The International Journal of Biochemistry and Cell Biology, 2005, 37(10):2134-2146 [7]GEESINK G H, KUCHAY S, CHISHTI A H & KOOHMARRAIE M.(2006).l-calpain is essential for postmortem proteolysis of muscle proteins. Journal of Animal Science, 84, 2834–2840. [8]黄明, 黄峰, 黄继超, 等.内源性蛋白酶对宰后肌肉嫩化机制研究进展[J].中国农业科学, 2011, 44(15):3214-3222 [9]TAYLOR R P & BENJAMIN I J(2005).Small heat shock proteins: a new classification scheme in mammals. Journal of Molecular and Cellular Cardiology, 38, 433–444. [10]LOMIWES D, HURST S M, DOBBIE P.et al. (2014b). The protection of bovine skeletal myofibrils from proteolytic damage post mortem by small heat shock proteins. Meat Science, 97, 548–557 [11]MORZEL M, TERLOUW C, CHAMBON C, Micol D.& Picard, B. (2008). Muscle proteome and meat eating qualities of Longissimus thoracis of “Blonde d’Aquitaine” young bulls: A central role of HSP27 isoforms. Meat Science, 78, 297–304. [12]LOMIWES D, FAROUK M M, FROST D A, et al.(2013). Small heat shock proteins and toughness in intermediate pHu beef. Meat Science, 95, 472–479. [13]CONTRERAS-CASTILLO C J, LOMIWES D, Wu G, FROST D.& FAROUK M M. (2016). The effect of electrical stimulation on post mortem myofibrillar protein degradation and small heat shock protein kinetics in bull beef. Meat Science, 113, 65–72. [14] VOSS O H, BATAR S, KOLATTUKUDY S J, et al.(2007). Binding of caspase-3 prodomain to heat shock protein 27 regulates monocyte apoptosis by inhibiting caspase-3 proteolytic activation. Journal of Biological Chemistry, 282, 25088–25099. [15] DING Z J, HUANG F, ZHANG C, et al.Effect of heat shock protein 27 on the invitro degradation of myofibrils by caspase‐3 and μ‐calpain[J]. International Journal of Food Science & Technology, 2018, 53(Supplement 1). [16]孙志昶, 余群力, 韩玲, 张文华, 杨勤.宰后牦牛肉骨骼肌细胞凋亡过程研究[J].肉类研究, 2014, 28(05):11-15 [17]贾青.细胞凋亡酶-3及其抑制剂对宰后牦牛肉品质变化的影响[D].甘肃农业大学, 2016. [18]XIA X, KONG B, LIU Q, et al.Physicochemical change and protein oxidation in porcine longissimus dorsi as influenced by different freeze-thaw cycles[J].Meat Science, 2009, 83(2):239- [19]AGYARE K K, XIONG Y L, ADDO K.Influence of salt and pH on the solubility and structural characteristics of transglutaminase-treated wheat gluten hydrolysate ☆[J].Food Chemistry, 2007, 107(3):1131-1137 [20]黄峰.细胞凋亡效应酶在牛肉成熟过程中的作用机制研究[D].南京农业大学, 2012. [21]KEMP C M, KING D A, SHACKELFORD S D, et al.The caspase proteolytic system in callipyge and normal lambs in longissimus,semimembranosus,and infraspinatus muscles during postmortem storage[J].Journal of Animal Science, 2009, 87(9):2943-2951 [22]SAMEJIMA, K., & WOLFE, F. H. (1976). Degradation of myofibrillar protein components during postmortem aging of chicken muscle. Journal of Food Science, 41(2), 250-254. [23]HUANG M，HUANG F，XUE M，et al.The effect of active caspase-3 on degradation of chicken myofibrillar proteins and structure of myofibrils[J].Food Chemistry, 2011, 64(12):1281- [24]WANG L L, YU Q L, HAN L, et al.Study on the effect of reactive oxygen species-mediated oxidative stress on the activation of mitochondrial apoptosis and the tenderness of yak meat. Food Chemistry, 2018, 244, 394-402. [25]郭有锋, 陈胜, 肖成峰, 等.热应激蛋白在苯致小鼠骨髓细胞凋亡中的作用[J].工业卫生与职业病, 2003, 29(1):17-19 [26]RANE, M.J. Heat Shock Protein 27 Controls Apoptosis by Regulating Akt Activation[J].Journal of Biological Chemistry, 2003, 278(30):27828-27835 [27]HUFF-ONERFAN E, MITSUHASHI T & BEEKMAN D D.(1996). Proteolysis of specific muscle structural proteins by m-Calpain at low pH and temperature is similar to degradation in postmortem bovine muscle. Journal of Animal Science, 74, 993–1008. [28]NOGUCHI K K, WALLS K C, WOZNIAK D F.2008. Acute neonatal glucocorticoid exposure produces selective and rapid cerebellar neural progenitor cell apoptotic death[J]. Cell death and differentiation, 15:1582-1592. [29]BEMARD C, CASSAR-MALEK I, LE C M, et al.New indicators of beef sensory quality revealed by expression of specific genes[J].Journal of Agricultural and Food Chemistry, 2007, 55(13):5229-5237 [30]BALAN P, KIM Y H, BLIJENBURG R.Small heat shock protein degradation could be an indicator of the extent of myofibrillar protein degradation[J].Meat Science, 2014, 97(2):220-222 [31]DELLA M A, MARINO R, SANTILLO A, et al.Proteomic approach to investigate the impact of different dietary supplementation on lamb meat tenderness. Meat Science, 2017, 131, 74-81.