大鼠脊髓损伤节段线粒体自噬蛋白及凋亡因子的表达
发布时间：2019-08-08 12:08
【摘要】：目的 :观察脊髓损伤节段线粒体自噬蛋白与凋亡蛋白的表达情况。方法 :将50只清洁级SD雄性大鼠随机分为损伤组(39只)和对照组(11只),损伤组应用Allen′s法建立大鼠脊髓损伤模型,并分为5个不同时间点(损伤后1h、6h、24h、48h、72h),每个时间点7只,在损伤不同时间点处死大鼠,并截取损伤区域约1cm长脊髓节段,通过Western blot检测线粒体自噬蛋白(LC3、NIX)及凋亡蛋白(BNIP3、cleaved caspase-3、cytochrome c)表达水平;另取4只大鼠在损伤后24h截取损伤组织,用透射电镜观察损伤后神经元自噬体结构。对照组不做任何处理,其中7只大鼠用于Western blot分析,4只用于电镜检测。结果:对照组及损伤后1h、6h、24h、48h、72h脊髓损伤组大鼠LC3表达水平分别为0.2893±0.0325、0.3002±0.0474、0.3943±0.1154、0.4818±0.0426、0.5430±0.0865、0.5790±0.0892;NIX表达分别为0.1392±0.0171、0.1431±0.0325、0.1955±0.1379、0.3841±0.1136、0.4043±0.1059、0.4506±0.0174;BNIP3表达水平分别为0.1354±0.0547、0.1896±0.1264、0.2654±0.1341、0.7220±0.1030、0.4713±0.1041、0.3975±0.1505;cleaved caspase-3表达水平分别为0.2806±0.0999、0.4158±0.1137、0.7865±0.4056、0.9354±0.2659、1.0152±0.3441、1.1608±0.2488;细胞色素C表达水平分别为0.1489±0.0300、0.2196±0.0762、0.3162±0.1656、0.4456±0.1180、0.5407±0.1029、0.5812±0.1388。LC3、NIX、cleaved caspase-3和细胞色素C在24h、48h、72h与对照组比较明显增加(P0.05);BNIP3在24h、48h与对照组比较明显增加(P0.05)。进一步通过透射电镜观察损伤后24h细胞超微结构,可观察到双层膜结构自噬体,其内包裹有受损线粒体等细胞器结构。结论:大鼠急性脊髓损伤后可能通过促进NIX蛋白与LC3结合诱导线粒体自噬,减少凋亡水平,从而在脊髓损伤修复中发挥保护作用。
[Abstract]:Objective: to observe the expression of mitochondrial autophagy protein and apoptotic protein in spinal cord injury segment. Methods: fifty male SD rats of clean grade were randomly divided into injury group (n = 39) and control group (n = 11). The spinal cord injury model was established by Allen's method and divided into 5 different time points (1 h, 6 h, 48 h, 72 h after injury). At each time point, 7 rats were killed at different time points of injury, and the long spinal cord segments of 1cm in the injured area were intercepted. The mitochondrial autophagin (LC3,) was detected by Western blot. The expression level of NIX and apoptosis protein (BNIP3,cleaved caspase-3,cytochrome c). The injured tissues of 4 rats were intercepted 24 hours after injury, and the autophagy structure of neurons after injury was observed by transmission electron microscope. No treatment was performed in the control group, of which 7 rats were used for Western blot analysis and 4 rats for electron microscopic examination. 缁撴灉:瀵圭収缁勫強鎹熶激鍚，
本文编号：2524356
[Abstract]:Objective: to observe the expression of mitochondrial autophagy protein and apoptotic protein in spinal cord injury segment. Methods: fifty male SD rats of clean grade were randomly divided into injury group (n = 39) and control group (n = 11). The spinal cord injury model was established by Allen's method and divided into 5 different time points (1 h, 6 h, 48 h, 72 h after injury). At each time point, 7 rats were killed at different time points of injury, and the long spinal cord segments of 1cm in the injured area were intercepted. The mitochondrial autophagin (LC3,) was detected by Western blot. The expression level of NIX and apoptosis protein (BNIP3,cleaved caspase-3,cytochrome c). The injured tissues of 4 rats were intercepted 24 hours after injury, and the autophagy structure of neurons after injury was observed by transmission electron microscope. No treatment was performed in the control group, of which 7 rats were used for Western blot analysis and 4 rats for electron microscopic examination. 缁撴灉:瀵圭収缁勫強鎹熶激鍚，
本文编号：2524356
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