Yang Ping, Ying Da-Jun, Song Lin, Sun Jian-Sen
Department of Anatomy, Third Military Medical University, Chongqing 400038.
Sheng Li Xue Bao. 2003 Aug 25;55(4):428-34.
The aim of this study was to investigate the protective effect of exogenous bcl-2 on spinal cord motoneurons following sciatic nerve axotomy. After epineurium suturing, sense bcl-2 (Ad/s-bcl-2), antisense bcl-2 (Ad/as-bcl-2), or reporter gene lacZ (Ad/lacZ) recombinant adenovirus or NS was injected into the sciatic nerve 0.5 cm distant from the sutured point respectively in different groups. The rats were transcardially perfused with 4% paraformaldehyde on postoperative 48 h, 7 d, 15 d and 30 d respectively and the spinal cords of L4 to L6 were harvested. X-gal staining, bcl-2 in situ hybridization and immunohistochemical staining, TUNEL (terminal deoxynucleotidyl transferase mediated dUTP nick end labeling) staining and AChE (acetyl cholinesterase) histochemical staining were used. We observed that the exogenous lacZ gene was expressed in the spinal cord of Ad/lacZ group, and sense bcl-2 significantly decreased the number of apoptotic motoneurons and the decreasing degree of AChE activity of the motoneurons in the spinal cord induced by sciatic nerve axotomy and accelerate AChE activity recovery. However, antisense bcl-2 increased the number of apoptotic motoneurons and the decreasing degree of AChE activity of the motoneurons in the spinal cord induced by sciatic nerve axotomy and prolonged AChE activity recovery. These results demonstrate that exogenous bcl-2 may protect motor neurons from injury induced by peripheral nerve axotomy.
本研究旨在探讨外源性bcl-2对坐骨神经切断后脊髓运动神经元的保护作用。在神经外膜缝合后,不同组分别于距缝合点0.5 cm处向坐骨神经注射正义bcl-2(Ad/s-bcl-2)、反义bcl-2(Ad/as-bcl-2)或报告基因lacZ(Ad/lacZ)重组腺病毒或生理盐水(NS)。分别在术后48 h、7 d、15 d和30 d经心脏灌注4%多聚甲醛处死大鼠,取L4至L6脊髓。采用X-gal染色、bcl-2原位杂交和免疫组化染色、TUNEL(末端脱氧核苷酸转移酶介导的dUTP缺口末端标记)染色及AChE(乙酰胆碱酯酶)组织化学染色。我们观察到Ad/lacZ组脊髓中有外源性lacZ基因表达,正义bcl-2可显著减少坐骨神经切断诱导的脊髓运动神经元凋亡数量及运动神经元AChE活性降低程度,并加速AChE活性恢复。然而,反义bcl-2增加了坐骨神经切断诱导的脊髓运动神经元凋亡数量及运动神经元AChE活性降低程度,并延长了AChE活性恢复时间。这些结果表明,外源性bcl-2可能保护运动神经元免受周围神经切断诱导的损伤。