中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (3): 542-552.doi: 10.4103/1673-5374.245481

• 原著:脊髓损伤修复保护与再生 • 上一篇    

黑色素瘤缺乏因子2在损伤脊髓组织中的表达和定位分布

  

  • 出版日期:2019-03-15 发布日期:2019-03-15
  • 基金资助:

    国家自然科学基金（81772321，81571194，81471277）；安徽省重点项目中国高校优秀人才项目（gxbjZD 2016071，2014H012）

Expression and localization of absent in melanoma 2 in the injured spinal cord

Sai-Nan Wang 1, 2, 3 , Xue-Yan Guo 1, 2, Jie Tang 3 , Shu-Qin Ding 1 , Lin Shen 2 , Rui Wang 2 , Shan-Feng Ma 2 , Jian-Guo Hu 1, 2 , He-Zuo Lü 1, 2, 3   

  1. 1 Clinical Laboratory, the First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui Province, China
    2 Anhui Key Laboratory of Tissue Transplantation, the First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui Province, China
    3 Department of Immunology, Bengbu Medical College, and Anhui Key Laboratory of Infection and Immunity at Bengbu Medical College, Bengbu, Anhui Province, China
  • Online:2019-03-15 Published:2019-03-15
  • Contact: He-Zuo Lü, MD, lhz233003@163.com; Jian-Guo Hu, PhD, jghu9200@163.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81772321 (to HZL), 81571194 (to HZL), 81471277 (to JGH); a grant from the Key Program of Anhui Province for Outstanding Talents in Universities in China, No. gxbjZD2016071 (to HZL), 2014H012 (to HZL).

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摘要：

在创伤性脑损伤的大脑组织中，黑素瘤缺乏因子2 (AIM2)已被证明可参与大脑组织神经元的焦亡性细胞死亡（pyroptotic cell death）。尽管脊髓损伤的病理生理机制与脑损伤相似，但目前脊髓损伤后AIM2的表达和细胞定位尚不十分清楚。为揭示此问题，实验采用重物高空坠落撞击法建立T9脊髓损伤模型大鼠，建模后分为脊髓损伤1 h，6 h，1 d，3 d和6 d组，同时设只暴露T9脊髓的假手术组作对照。(1)免疫印迹检测显示，在脊髓损伤1 h、6 h和1 d组间AIM2蛋白表达无显著性差异。AIM2在脊髓损伤1 h、6 h和1 d组中的表达显著高于假手术组、脊髓损伤3 d和7 d组；(2)采用双标记免疫荧光染色结果显示，在假手术组脊髓中存在NeuN+(神经元标记物)/ AIM2+，GFAP+(星形胶质细胞标记物)/AIM2+，CNPase+(少突胶质细胞标记物)/AIM2+和CD11b+(小胶质细胞标记物)/AIM2+细胞。脊髓损伤后各时间点大鼠损伤脊髓组织中AIM2可发现CD45 (白细胞标记物)/AIM2和CD68+(活化的小胶质细胞/巨噬细胞标记物)/ AIM2荧光双标细胞分布；(3)上述结果数据表明，在正常的脊髓中AIM2主要表达于神经元、星形胶质细胞、小胶质细胞和少突胶质细胞中。脊髓损伤后，AIM2表达随着白细胞的浸润、星形胶质细胞和小胶质细胞/巨噬细胞的活化而逐渐增加。

orcid: 0000-0002-3889-835X(He-Zuo Lü)
           0000-0002-9055-874X(Jian-Guo Hu)

关键词: 脊髓损伤, 黑素瘤缺乏因子2, 时空表达, 神经元, 星形胶质细胞, 少突胶质细胞, 浸润的白血病, 活化的小胶质细胞, 免疫印迹, 免疫组织化学, 神经再生

Abstract:

In traumatic brain injury, absent in melanoma 2 (AIM2) has been demonstrated to be involved in pyroptotic neuronal cell death. Al¬though the pathophysiological mechanism of spinal cord injury is similar to that of brain injury, the expression and cellular localization of AIM2 after spinal cord injury is still not very clear. In the present study, we used a rat model of T9 spinal cord contusive injury, produced using the weight drop method. The rats were randomly divided into 1-hour, 6-hour, 1-day, 3-day and 6-day (post-injury time points) groups. Sham-operated rats only received laminectomy at T9 without contusive injury. Western blot assay revealed that the expression levels of AIM2 were not significantly different among the 1-hour, 6-hour and 1-day groups. The expression levels of AIM2 were markedly higher in the 1-hour, 6-hour and 1-day groups compared with the sham, 3-day and 7-day groups. Double immunofluorescence staining demonstrated that AIM2 was expressed by NeuN+ (neurons), GFAP+ (astrocytes), CNPase+ (oligodendrocytes) and CD11b+ (microglia) cells in the sham-operated spinal cord. In rats with spinal cord injury, AIM2 was also found in CD45+ (leukocytes) and CD68+ (activated microglia/macrophages) cells in the spinal cord at all time points. These findings indicate that AIM2 is mainly expressed in neurons, astro¬cytes, microglia and oligodendrocytes in the normal spinal cord, and that after spinal cord injury, its expression increases because of the infiltration of leukocytes and the activation of astrocytes and microglia/macrophages.

Key words: nerve regeneration, spinal cord injury, absent in melanoma 2, spatio-temporal expression, neurons, astrocytes, oligodendrocytes, infiltrated leukocytes, activated microglia, western blot assay, immunohistochemistry, neural regeneration