大剂量X射线辐照对离体大鼠脊髓神经元凋亡的影响  

X-ray irradiation induces apoptosis of spinal neurons of SD rats in vitro

作　　者：王志军[1] 李婷[1] 司胜斌 丁娟[3] 夏新舍[1] 赵薇 Wang Zhijun;Li Ting;Si Shengbin;Ding Juan;Xia Xinshe;Zhao Wei(General Hospital;Key Laboratory of Reproduction and Heredity;Basic Medicine College,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院 [2]宁夏医科大学生育力保持教育部重点实验室 [3]宁夏医科大学基础医学院,银川750004

出　　处：《神经解剖学杂志》2018年第5期610-614,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81560510;81360408;81460478)

摘　　要：目的:研究X射线辐照对大鼠原代脊髓神经元细胞的损伤效应。方法:制备SD大鼠脊髓原代神经元细胞,神经元标志物神经元核抗原(Neu N)免疫荧光染色鉴定其纯度。给予0、3、6和9 Gy等距离X射线辐照,二甲基噻唑-二苯基四氮唑溴盐(MTT)比色法检测神经元存活率,乳酸脱氢酶(LDH)漏出率评价细胞膜完整性。Western Blot检测DNA损伤反应蛋白-聚(ADP核糖)聚合酶1(PARP1)完整型和裂解型蛋白的表达水平以及胞浆胞核蛋白中凋亡诱导因子(AIF)的表达变化。结果:大鼠脊髓原代90%Neu N染色阳性,提示神经元纯度达90%以上。原代神经元细胞接受0、3、6和9 Gy等距离X射线辐照后48 h,9 Gy辐照使得细胞存活率显著降低(P <0. 05)及LDH漏出率升高(P <0. 05)。6 Gy辐照后神经元细胞中完整型PARP1表达显著升高(P <0. 05),而9 Gy辐照后PARP1水解明显加剧(P <0. 01),提示凋亡增加。6 Gy及以上剂量辐照后神经元细胞核蛋白中AIF显著增多(P <0. 05)。结论:大剂量X射线辐照破坏了大鼠脊髓神经元细胞膜完整性,引起DNA损伤,诱导了caspase依赖及非caspase依赖的细胞凋亡。Objective： To study the effect of X-ray irradiation on spinal neurons in SD rats.Methods： Primary spinal cord neurons from SD rats were identified using an immunofluorescent staining with Neu N antibody.After primary neurons were exposed to the X-ray with gradient doses（ 0,3,6 or 9 Gy）,survival rate and the membrane integrity of the neurons were evaluated by MTT and the leakage rate of LDH,respectively.Expression of whole and cleaved Poly[ADP-ribose]polymerase 1（ PARP1） were analyzed by Western Blot.Results： More than 90% of the primary neurons belonged to the Neu N positive cells.The survival rate was decreased（ P〈0.05） and the leakage rate of LDH was increased（ P〈0.05） in neurons with a 9 Gy dose of X-ray irradiation.48 hours after the neurons with a 6 Gy-X-rayirradiation,expression level of whole PARP1 in neurons were increased（ P〈0.05）.A 9 Gy irradiation promoted whole PARP1 to cleave and inactivate（ P〈0.01）.6 Gy and 9 Gy irradiations increased nucleic AIF expression of neurons（ P〈0.01）.Conclusion： X ray destroys the membrane integrity of rat spinal neurons and induces caspase-dependent and non-caspase dependent apoptosis of neurons.

关 键 词：聚(ADP核糖)聚合酶1 早期凋亡诱导因子 凋亡 原代脊髓神经元 X射线 

分 类 号：R651.2[医药卫生—外科学]