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目的观察成年肌萎缩脊髓侧索硬化症(ALS)转基因模型鼠脊髓内源性细胞的增殖和存活情况。方法对ALS转基因鼠进行BrdU注射,分别于不同时间点取材,冷冻切片,应用免疫荧光染色技术检测脊髓内细胞的增殖、分布和存活情况。结果成年ALS鼠脊髓的中央管、灰质、白质均可检测到BrdU阳性细胞,经常可检测到形状相似、成对出现的细胞。在灰质内,BrdU阳性细胞主要分布于脊髓前角。在白质内,BrdU阳性细胞散在分布。前角ALS鼠脊髓内BrdU阳性细胞数量较野生型鼠多。BrdU末次注射后1d、14d均可检测到BrdU阳性细胞,但细胞数量逐渐减少,28d ALS鼠脊髓内仍可检测到少量BrdU阳性细胞,主要分布于中央管部位。增殖细胞核抗原(PCNA)阳性细胞的分布和变化规律与BrdU阳性细胞一致。在95d龄ALS鼠(BrdU末次注射1d后),脊髓中Nestin阳性细胞较野生型鼠多,多数Nestin阳性细胞分布于脊髓前角,某些细胞呈BrdU/Nestin双标阳性。在BrdU注射后14d、28d脊髓中,未检测到BrdU/Nestin双标阳性细胞。结论神经退行性病变可激活ALS转基因鼠脊髓的细胞增殖,且细胞存活可达28d。
Objective To observe the proliferation and survival of endogenous spinal cord cells in adult amyotrophic lateral sclerosis (ALS) transgenic mice. Methods The ALS transgenic mice were injected with BrdU. The cells were harvested at different time points and frozen sectioned. The proliferation, distribution and survival of spinal cord cells were detected by immunofluorescence staining. Results BrdU positive cells were detected in the central tube, gray matter and white matter of adult ALS rats spinal cord, and cells with similar shape and paired appearance were always detected. Within the gray matter, BrdU positive cells are mainly distributed in the anterior horn of the spinal cord. In the white matter, BrdU-positive cells scattered. The number of BrdU positive cells in spinal cord of ALS rats was higher than that of wild type mice. BrdU positive cells could be detected on the 1st and 14th day after the last injection of BrdU, but the number of BrdU positive cells gradually decreased. A small amount of BrdU positive cells could still be detected in the spinal cord of the 28th ALS mice, which mainly distributed in the central tube. The distribution and change of proliferating cell nuclear antigen (PCNA) positive cells were consistent with that of BrdU positive cells. Nestin positive cells in the spinal cord were more than those in the wild type mice at the 95th day after the BrdU injection for 1 day. Most of the Nestin positive cells were located in the anterior horn of the spinal cord and some were positive for BrdU / Nestin double staining. BrdU / Nestin double positive cells were not detected in the spinal cord on the 14th day and the 28th day after BrdU injection. Conclusion Neurodegenerative diseases can activate the proliferation of spinal cord in ALS transgenic mice, and the cell survival rate can reach 28 days.