摘要
目的探讨自噬与凋亡相关因子在人退变髓核中的共表达及其意义。方法收集2011年6月至2012年6月75例中山大学孙逸仙纪念医院住院患者腰椎间盘髓核标本，其中正常组10例来自腰椎骨折或肿瘤未侵犯椎间盘者，退变组65例来自腰椎间盘突出症或腰椎滑脱的手术患者。所有病例均行MRI检查并按Pfirrmann分级标准进行分级，其中Ⅰ／Ⅱ级10例，Ⅲ级32例，Ⅳ级22例，Ⅴ级11例。苏木素-伊红（HE）染色检测椎间盘组织的退变程度，原位末端转移酶标记（TUNEL）检测髓核细胞凋亡，Western blot检测自噬相关蛋白Beclin-1、Unc51样激酶-1（ULK-1）、微管相关蛋白1轻链3（LC3）、凋亡相关蛋白多腺苷二磷酸多聚酶（PARP）p85及白噬、凋亡相关蛋白B细胞淋巴瘤／白血病-2／腺病E1B相互作用相似蛋白3（BNIP3L）的表达。结果随着椎间盘退变程度的增加，细胞凋亡的发生率及其相关蛋白PARPp85表达增加（9．35±1．91、23．40±3．40、26．85±2．85和35．80±3．43，P〈0．05；0．30±0．04、0．80±0．22、0．88±0．06和1．07±0．05，P〈0．05），但自噬相关蛋白Beclin-1、ULK-1、LC3先增加后降低（0．40±0．24、1．08±0．02、1．22±0．13和0．89±0．06，P〈0．05；0．65±0．10、0．95±0．17、0．89±0．13和0．51±0．17，P〈0．05；0．20±0．09、0．65±0．10、0．69±0．27和0．44±0．10，P〈0．05）。BNIP3L蛋白亦呈先增加后降低的趋势（0．57±0．19、1．19±0．26、0．99±0．12和0．16±0．05，P〈0．05）。结论在椎间盘退变进程中，自噬与凋亡表达不同，作用亦可能不同。
Objective To explore the co-expression of autophagy and apoptosis related factors in human degenerative nucleus pulposus. Methods From June, 2011 to June, 2012, 75 lumbar nucleus pulposus were collected. According to Pfirrmann classification, 10 of them, grade Ⅰ / Ⅱ, were from the cases with lumbar fracture or spinal tumor, 65, including 32 with grade m, 22 with grade Ⅳ and Ⅱ with grade V, were from the cases with lumbar disc herniation or spondylolisthesis. Hematoxylin and eosin （HE） stain was used to verify the grade of intervertebral disc degeneration and apoptosis was detected by TdT-mediated dUTP nick end labeling （TUNEL）. The expression of autophagy-related proteins, such as microtubule-assoeiated protein 1 light chain 3 （ LC3）, Unc51-1ikekinase-1 （ ULK-1 ） and Beclin-1, and the apoptosis-related protein poly adenosine diphosphate-ribose polymerase （PARP） p85 were detected by Western blotting. The apoptosis and autophagy-related proteins B-cell lymphoma/adenovirus E1B 19 000 interacting protein 3-like（ BNIP3L）, were also measured by Western blotting. Results With the disc de- generation increasing, the rate of nucleus pulposus cell apoptosis and the expression of apoptosis-related protein ascended （TUNEL： 9.35±1.91, 23.40±3.40, 26.85 ±2.85 and 35.80 ±3.43, P〈0.05; PARP p85 ： 0. 30 ± O. 04, 0. 80 ± 0. 22, 0. 88 ± 0. 06 and 1.07 ± 0. 05, P 〈 0. 05 ）. However, the autoph- agy-related factors just increased and then decreased with the disc degeneration increasing （Beclin-1 ： 0.40±0.24, 1.08 ±0.02, 1.22 ±0. 13 and 0.89 ±0.06, P〈0.05; ULK-1 ： 0.65 ±0. 10, 0.95 ± 0. 17, 0.89±0.13 and 0.51 ±0.17, P〈0.05; LC3：0.20 ±0.09, 0.65 ±0. 10, 0.69 ±0.27 and 0. 44 ±0. 10, P 〈0.05）. And the expression of BNIP3L protein increased and then decreased with the disc degeneration increasing （0. 57 ± 0. 19, 1.19 ± 0. 26, 0. 99 ± 0. 12 and 0. 16 ± 0. 05, P 〈 0. 05）. Conclusion In the process of intervertebral disk degeneration, the expressions of
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第5期1062-1064,共3页
Chinese Journal of Experimental Surgery
基金
基金项目：国家自然科学基金资助项目（81101385）
广东省自然科学基金资助项目（10151008901000084）
广东省科技计划资助项目（20128031800359、20128031800360）
广东省大学生创新实验项目（1055811196）
关键词
椎间盘退变
髓核
自噬
凋亡
B细胞淋巴瘤
白血病-2
腺病E1B相互作
用相似蛋白3
Intervertebral disc degeneration
Nucleus pulposus
Autophagy
Apoptosis
B-cell lymphoma/Leukemia-2/adnovirus E1B 19 000 interacting protein 3-like
