张力，崔宝娟，曾凡硕，黄来刚，张琪，孙敏，刘本玲，李强，王道清，孙强三.自噬相关基因及蛋白在急性脊髓损伤后大鼠膀胱表达的实验研究[J].中华物理医学与康复杂志,2018,40(6):401-406
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| 自噬相关基因及蛋白在急性脊髓损伤后大鼠膀胱表达的实验研究 |
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| DOI： |
| 中文关键词: 自噬 急性期脊髓损伤 膀胱 微管相关蛋白1轻链3 |
| 英文关键词: Autophagy Spinal cord injury Bladder Microtubule-associated protein 1 light chain 3 |
| 基金项目:山东省自然科学基金(ZR2014HL080)，山东省自然科学基金(ZR2014HP031），山东省医药卫生科技发展计划(2015WS0310) |
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| 中文摘要: |
| 目的 观察自噬相关基因、蛋白在急性期脊髓损伤大鼠膀胱平滑肌组织中的表达。 方法 选取24只雄性Wistar大鼠并随机分为模型组（12只）及对照组（12只）。模型组采用改良Allen′s法建立脊髓损伤模型，对照组仅予以椎板切除处理。于术后6h对2组大鼠进行BBB运动评分，采用尼氏染色观察2组大鼠脊髓形态学变化，采用Western blotting和免疫荧光染色法检测膀胱平滑肌细胞微管相关蛋白1轻链3(LC3)及P62表达，采用RT-PCR检测膀胱平滑肌自噬相关基因LC3、P62、Beclin1 mRNA表达。 结果 模型组大鼠下肢运动功能BBB评分较对照组明显降低（P<0.05）。尼氏染色观察可见模型组神经元及尼氏小体数量减少；Western blotting染色提示模型组LC3-Ⅱ表达较对照组增高（P<0.05），P62表达较对照组降低（P<0.05)。免疫荧光染色法提示模型组膀胱平滑肌LC3阳性细胞率较对照组增高（P<0.05)，P62阳性细胞率较对照组降低（P<0.05）。RT-PCR检测发现模型组LC3及Beclin1 mRNA水平较对照组升高(P<0.05），P62 mRNA水平较对照组降低(P<0.05）。 结论 自噬在急性脊髓损伤大鼠膀胱平滑肌中表达活跃，可能与脊髓损伤后神经源性膀胱发病机制有关。 |
| 英文摘要: |
| Objective To observe the expression of autophagy-related genes and proteins in the smooth bladder muscle of rats after spinal cord injury (SPI). Methods Twenty-four male Wistar rats were randomly and evenly divided into a model group and a control group. The model group had SPI induced using the modified Allen′s method, while the control group was only given laminectomy. Six hours after the operation, the Basso Beattle Bresnahan (BBB) locomotor rating scale was used to evaluate the rats′ hindlimb locomotor function. Nissl staining was used to observe the morphological changes in their spinal cords, while Western blotting and immunofluorescence staining were employed to assess the expression of microtubule-associated protein 1 light chain 3 (LC3) and protein 62 (P62). The expression of autography gene Beclin1 mRNA was determined using a reverse transcription polymerase chain reaction (RT-PCR). Results The average BBB score in the model group was significantly lower than in the control group. After Nissl′s staining, a decreased number of neurons and Nissl bodies was observed. Western blotting showed that the expression of LC3-II had increased significantly and that of P62 had decreased significantly in the model group compared with the control group. The immunofluorescence staining showed LC3 and P62 dots in the bladders′ smooth muscle cells. RT-PCR detected significantly higher LC3 and Beclin1 mRNA levels in the model group than in the control group; in contrast the average P62 mRNA level was significantly lower. Conclusions Autophagy was activated in rats′ bladder muscles after SPI. That may be related to the pathogenesis of a neurogenic bladder after spinal cord injury. |
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