| 杜傲男,徐静,何燕,等.神经源性分化因子对小鼠放射性肠损伤的治疗作用[J].中华放射医学与防护杂志,2015,35(1):45-48.Du Aonan,Xu Jing,He Yan,et al.The therapeutic effect of NeuroD on radiation-induced intestinal injury in mice[J].Chin J Radiol Med Prot,2015,35(1):45-48 |
| 神经源性分化因子对小鼠放射性肠损伤的治疗作用 |
| The therapeutic effect of NeuroD on radiation-induced intestinal injury in mice |
| 投稿时间：2014-08-07 |
| DOI：10.3760/cma.j.issn.0254-5098.2015.01.008 |
| 中文关键词: 电离辐射 神经源性分化因子 辐射损伤 放射性肠损伤 |
| 英文关键词:Ionizing radiation Neurogenic differentiation (NeuroD) Radiation injury Radiation-induced intestinal injuries |
| 基金项目:国家自然科学基金 (81172597, 31300694, 81472917) |
| 作者 | 单位 | E-mail | | 杜傲男 | 苏州大学医学部放射医学与防护学院
江苏高校放射医学协同创新中心, 江苏省放射医学与防护重点实验室, 苏州 215123 | | | 徐静 | 苏州大学医学部放射医学与防护学院
江苏高校放射医学协同创新中心, 江苏省放射医学与防护重点实验室, 苏州 215123 | | | 何燕 | 苏州大学医学部放射医学与防护学院
江苏高校放射医学协同创新中心, 江苏省放射医学与防护重点实验室, 苏州 215123 | | | 李明 | 苏州大学医学部放射医学与防护学院
江苏高校放射医学协同创新中心, 江苏省放射医学与防护重点实验室, 苏州 215123 | | | 朱巍 | 苏州大学医学部放射医学与防护学院
江苏高校放射医学协同创新中心, 江苏省放射医学与防护重点实验室, 苏州 215123 | | | 张旭光 | 徐州市肿瘤医院 | | | 曹建平 | 苏州大学医学部放射医学与防护学院
江苏高校放射医学协同创新中心, 江苏省放射医学与防护重点实验室, 苏州 215123 | jpcao@suda.edu.cn |
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| 中文摘要: |
| 目的 研究神经源性分化因子(NeuroD)对放射性肠损伤的治疗作用.方法 采用原核表达系统表达、纯化NeuroD与绿色荧光蛋白(EGFP)的融合蛋白(NeuroD-EGFP融合蛋白),利用倒置荧光显微镜观察NeuroD-EGFP融合蛋白的体外进入细胞的效率.40只C57BL/6J小鼠采用随机数字表法分为正常对照组、磷酸盐缓冲液(PBS)组、EGFP组、NeuroD-EGFP融合蛋白组,每组10只.除正常对照组外,其余3组动物经9 Gy γ射线全身照射后,观察NeuroD-EGFP融合蛋白在小肠上皮细胞内的分布情况和NeuroD对放射性肠损伤的治疗作用.结果 纯化得到了NeuroD-EGFP融合蛋白.细胞上清中加入NeuroD-EGFP融合蛋白后,可见绿色荧光聚集在细胞内部,表明其可穿过细胞膜进入细胞内.小鼠腹腔注射NeuroD-EGFP融合蛋白5 h后,小肠绒毛上皮细胞内部有绿色荧光聚集.小鼠照射后3.5 d,NeuroD-EGFP组小鼠绒毛高度高于PBS及EGFP组(F=49.49,P<0.01),隐窝深度及隐窝数目大于PBS及EGFP组(F=16.72、10.32,P<0.01).结论 NeuroD蛋白可促进放射后肠道绒毛及隐窝的修复,对放射性肠损伤具有治疗作用. |
| 英文摘要: |
| Objective To evaluate the therapeutic effect of NeuroD protein on radiation-induced intestinal injuries.Methods The expression and purification of NeuroD-enhanced green fluorescent protein (EGFP) fusion protein was performed in prokaryotic expression system. The efficiency of the fusion protein transduction into cells was monitored under fluorescence microscope. C57BL/6J mice were randomly divided into four groups with 10 mice in each group: normal control group, PBS group, EGFP group, and NeuroD-EGFP group. Besides the normal control group, the other three groups of mice received 9 Gy γ-ray total body irradiation. Intestinal tissues were collected, frozen sections were prepared to monitor the distribution of NeuroD in mice intestinal tract under fluorescence microscope, and pathological sections were prepared for H&E staining to evaluate the therapeutic effect of NeuroD protein. Results The NeuroD-EGFP fusion protein was purified by Ni-NTA column and verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Visible green fluorescence gathered within the cells after NeuroD-EGFP fusion protein was added in the culture medium, suggesting that NeuroD-EGFP could penetrate the cell membrane into the cells. Five hours after intraperitoneal injection of NeuroD-EGFP, visible green fluorescence gathered within the intestinal epithelial cells in villi. At 3.5 d after irradiation, NeuroD-EGFP treated mice showed significantly higher villus (F=49.49, P<0.01) and crypt depth (F=16.72, P<0.01) and more crypts per circumference (F=10.32, P<0.01) compared with PBS and EGFP groups. Conclusion NeuroD protein can accelerate the post-irradiation recovery of injured villi and crypt of intestinal tract and could be used to treat radiation-induced intestinal injuries. |
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