脂联素修复鸡骨骼肌细胞氧化损伤的分子机制研究

发布时间：2018-04-01 23:28

本文选题：肉鸡　切入点：脂联素　出处：《西北农林科技大学》2017年硕士论文

【摘要】：本研究通过构建鸡骨骼肌细胞氧化应激模型,探讨脂联素在p53、溶酶体、线粒体等的参与下,对肉鸡骨骼肌细胞氧化损伤的修复机制,为肉鸡抗应激高效养殖提供依据。试验结果如下:1分离培养肉鸡骨骼肌卫星细胞并鉴定,过氧化氢处理构建肉鸡骨骼肌氧化应激模型,分析氧化应激和细胞活力相关指标。结果表明:(1)DESMIN免疫荧光鉴定培养获得的细胞为骨骼肌卫星细胞;(2)H2O2处理获得的卫星细胞,ROS、MDA含量水平和TUNEL染色细胞凋亡率显著升高,细胞活力、SOD和GSH-Px酶活力单位显著降低(P0.05),表明过氧化氢处理构建的骨骼肌卫星细胞氧化应激模型成功。2脂联素处理肉鸡骨骼肌氧化应激模型卫星细胞,分析细胞内溶酶体和线粒体指标的变化。结果表明:(1)脂联素处理组细胞ROS、MDA含量水平和凋亡率显著降低,细胞活力、SOD和GSH-Px酶活力单位显著升高(P0.05);(2)脂联素处理组显著改善溶酶体和线粒体的膜通透性,Bax、Cyt-c、Caspase 8、Cathepsin B、Chymosin基因和蛋白水平均显著降低,Bcl-2基因和蛋白水平显著升高(P0.05)。表明脂联素通过改善细胞溶酶体和线粒体功能来修复骨骼肌细胞的氧化损伤。3 p53抑制剂PFT-α与脂联素共同处理氧化应激模型细胞,研究p53参与脂联素修复氧化应激的机制。结果表明:与脂联素处理氧化应激模型组相比,(1)PFT-α进一步显著降低了细胞ROS、MDA含量水平和TUNEL凋亡率,促进细胞活力、SOD和GSH-Px酶活力提高(P0.05);(2)PFT-α改善溶酶体和线粒体的膜通透性,抑制Bax、Cyt-c、Caspase 8、Cathepsin B、Chymosin基因和蛋白表达水平,促进Bcl-2基因和蛋白表达(P0.05)。抑制p53表达,可进一步促进脂联素修复骨骼肌细胞氧化损伤的作用。综上所述,脂联素可通过抑制p53的表达,改善细胞溶酶体和线粒体膜通透性、恢复膜电位、抑制凋亡基因和蛋白的表达、以及细胞凋亡进程等途径,修复肉鸡骨骼肌细胞的氧化损伤。
[Abstract]:In this study, the oxidative stress model of chicken skeletal muscle cells was established to explore the repair mechanism of oxidative damage of skeletal muscle cells induced by adiponectin with the participation of p53, lysosome and mitochondria, so as to provide evidence for anti-stress and high-efficiency breeding of broilers.The results were as follows: firstly, the skeletal muscle satellite cells of broilers were isolated and identified. The oxidative stress model of broiler skeletal muscle was constructed by hydrogen peroxide treatment, and the related indexes of oxidative stress and cell viability were analyzed.Sod and GSH-Px activity units significantly decreased P0.05, which indicated that oxidative stress model of skeletal muscle satellite cells constructed by hydrogen peroxide treatment was successfully treated with adiponectin in broiler skeletal muscle oxidative stress model satellite cells.The changes of intracellular lysosomes and mitochondria were analyzed.The results showed that the content of MDA and apoptosis rate of ROSN in adiponectin treated group were significantly lower than those in control group.It is suggested that adiponectin can repair oxidative damage of skeletal muscle cells by improving the function of lysosome and mitochondria. 3. PFT- 伪, a p53 inhibitor, co-treated oxidative stress model cells with adiponectin, and studied the mechanism of p53 involved in the repair of oxidative stress by adiponectin.The results showed that compared with adiponectin treated oxidative stress model group, PFT- 伪 further decreased the content of rosid and TUNEL apoptosis, and promoted the activity of sod and GSH-Px. P0.05PFT- 伪 improved the membrane permeability of lysosomes and mitochondria.The expression level of Chymosin gene and protein was inhibited and the expression of Bcl-2 gene and protein was promoted by inhibiting the expression of Caspase 8 and Cathepsin Chymosin, and promoting the expression of Bcl-2 gene and protein.Inhibition of p53 expression could further promote the effect of adiponectin on the repair of oxidative injury of skeletal muscle cells.In conclusion, adiponectin can inhibit the expression of p53, improve the permeability of lysosome and mitochondria, restore membrane potential, inhibit the expression of apoptotic genes and proteins, and the process of apoptosis.Repair oxidative damage of skeletal muscle cells in broilers.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S831

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