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自噬在大鼠急性脊髓损伤肺组织中的变化

Changes of autophagy in rat lung tissue after acute spinal cord injury

摘要:

目的 探讨自噬在大鼠急性脊髓损伤(ASCI)肺组织中的变化及可能机制.方法 63只雌性SD大鼠按随机数字表法分为假手术组(21只)和脊髓损伤组(42只).采用改良Allen法以10 ×25 9·mm能量于T10节段打击脊髓,制备ASCI模型.于伤后6,12,24,48,72 h、1,2周取材.通过HE染色和TUNEL法观察肺组织损伤、细胞凋亡情况.采用Western blot法及免疫荧光检测肺组织的自噬变化及LC3-Ⅱ、P62、Beclin-1、白细胞介素(IL)-17A、Bcl-2表达水平. 结果 假手术组肺泡结构正常,无炎症及出血.脊髓损伤组于伤后12 h肺组织出现轻微损伤;肺泡间质增宽、炎性浸润、出血、肺泡塌陷等损伤于伤后24~72 h逐渐出现并加重.脊髓损伤组伤后24,72 h和1周凋亡细胞数分别为(551.22±135.94)个/视野、(905.11 ±92.64)个/视野和(141.78±30.86)个/视野,24 h和72 h较假手术组增多(P<0.05).脊髓损伤组LC3-Ⅱ相对表达量在伤后24~72 h较假手术组增高(P<0.05).与假手术组比较,脊髓损伤组P62相对表达量在伤后24 ~72 h上调(P<0.05);Beclin-1相对表达量伤后24 h升高(P<0.05),48~72 h出现下降(P<0.05);IL-17A相对表达量在伤后24 ~ 48 h升高(P<0.05);Bcl-2相对表达量伤后24~72 h升高(P<0.05). 结论 大鼠ASCI后肺组织自噬小体形成增多并出现蓄积,其发生可能与IL-17A通过上调Bcl-2从而增加Beclin-1与Bcl-2的相互作用,最终导致自噬被抑制.

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abstracts:

Objective To investigate the changes of autophagy in rat lung tissues after acute spinal cord injury (ASCI) and the possible mechanisms.Methods Sixty-three female SD rats were divided into sham operation group (n =21) and spinal cord injury group (n =42),according to the random number table.The modified Allen method with the impact energy of 10 × 25 g · mm at the T10 vertebra was used for preparation of ASCI models.The rats were sacrificed at 6,12,24,48,72 hours,1 and 2 weeks after injury.Lung tissue damage and apoptosis were detected by HE staining and TUNEL method.The changes of autophagy and expressions of LC3-Ⅱ,P62,Beclin-1,interleukin (IL)-17A and Bcl-2 in lung were detected by Western blot and immunofluorescence.Results The pulmonary alveoli maintained normal structure in sham operation group,with no inflammation or pulmonary hemorrhage.Slight lung tissue damages were observed in spinal cord injury group at 12 h postinjury.Alveolar stroma widening,inflammatory infiltration,hemorrhage,and alveolar collapse became ingravescent at 24-72 hours postinjury.Numbers of apoptotic cells in spinal cord injury group were 551.22 ± 135.94,905.11 ±92.64,and 141.78 ± 30.86 respectively at 24,72 hours and 1 week postinjury,and were significantly increased at 24 and 72 hours postinjury,compared with sham operation group (P < 0.05).Expression of LC3-II in spinal cord injury group was increased at 24-72 hours postinjury,compared with sham operation group (P < 0.05).Expression of P62 in spinal cord injury group was up regulated at 24-72 hours postinjury,compared with sham operation group (P < 0.05).Expression of Beclin-1 in spinal cord injury group was increased at 24 h postinjury and then dropped at 48-72 hours,compared with sham operation group (P < 0.05).Expression of IL-17A in spinal cord injury group was increased at 24-48 hours,compared with sham operation group (P < 0.05).Expression of Bcl-2 in spinal cord injury group was increased from 24 hours to 72 hours,compared with sham operation group (P < 0.05).Conclusion Autophagosome formation is increased and accumulated in the lung tissues after ASCI,which might be related to the increased interaction between Beclin-1 and Bcl-2 because of the up regulation of Bcl-2 by IL 17A,ultimately leading to the inhibition of autophagy.

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作者: 储瑞亮 [1] 汪九龄 [1] 毕杨 [1] 南国新 [1]
期刊: 《中华创伤杂志》2018年34卷3期 253-259页 ISTICPKUCSCD
栏目名称: 基础研究
DOI: 10.3760/cma.j.issn.1001-8050.2018.03.012
发布时间: 2018-05-18
基金项目:
国家自然科学基金 国家临床重点专科建设项目[国卫办医函(2013)544]National Natural Science Foundation of China National Key Specialty Construction of Clinical Projects
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