【摘要】：目的 :觀察脊髓損傷節(jié)段線粒體自噬蛋白與凋亡蛋白的表達(dá)情況。方法 :將50只清潔級SD雄性大鼠隨機分為損傷組(39只)和對照組(11只),損傷組應(yīng)用Allen′s法建立大鼠脊髓損傷模型,并分為5個不同時間點(損傷后1h、6h、24h、48h、72h),每個時間點7只,在損傷不同時間點處死大鼠,并截取損傷區(qū)域約1cm長脊髓節(jié)段,通過Western blot檢測線粒體自噬蛋白(LC3、NIX)及凋亡蛋白(BNIP3、cleaved caspase-3、cytochrome c)表達(dá)水平;另取4只大鼠在損傷后24h截取損傷組織,用透射電鏡觀察損傷后神經(jīng)元自噬體結(jié)構(gòu)。對照組不做任何處理,其中7只大鼠用于Western blot分析,4只用于電鏡檢測。結(jié)果:對照組及損傷后1h、6h、24h、48h、72h脊髓損傷組大鼠LC3表達(dá)水平分別為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表達(dá)分別為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表達(dá)水平分別為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表達(dá)水平分別為0.2806±0.0999、0.4158±0.1137、0.7865±0.4056、0.9354±0.2659、1.0152±0.3441、1.1608±0.2488;細(xì)胞色素C表達(dá)水平分別為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和細(xì)胞色素C在24h、48h、72h與對照組比較明顯增加(P0.05);BNIP3在24h、48h與對照組比較明顯增加(P0.05)。進(jìn)一步通過透射電鏡觀察損傷后24h細(xì)胞超微結(jié)構(gòu),可觀察到雙層膜結(jié)構(gòu)自噬體,其內(nèi)包裹有受損線粒體等細(xì)胞器結(jié)構(gòu)。結(jié)論:大鼠急性脊髓損傷后可能通過促進(jìn)NIX蛋白與LC3結(jié)合誘導(dǎo)線粒體自噬,減少凋亡水平,從而在脊髓損傷修復(fù)中發(fā)揮保護(hù)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. 緇撴灉:瀵圭収緇勫強鎹熶激鍚，

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