Ischemic postconditioning protects neuronal death caused by cerebral ischemia and reperfusion via attenuating protein aggregation.
Ischemic postconditioning protects neuronal death caused by cerebral ischemia and reperfusion via attenuating protein aggregation.
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缺血后处理通过减弱蛋白质聚集来保护脑缺血和再灌注引起的神经元死亡。
DOI：
10.7150/ijms.4878
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发表时间：
2012
影响因子：
3.6
通讯作者：
Ge P
中科院分区：
文献类型：
--
作者：
Liang J;Yao J;Wang G;Wang Y;Wang B;Ge P
目的：探讨缺血后处理对短暂缺血再灌注引起的蛋白质聚集的影响及其机制。方法：采用双血管闭塞法建立大鼠短暂性全脑缺血模型。缺血后处理组在缺血15 min后进行3个循环的30-s/30-s再灌注/钳夹。通过苏木精-伊红染色观察CA 1区的神经元死亡，并在光学显微镜下通过细胞计数评估活神经元的数量。以琥珀酰-LLVY-AMC为底物，体外测定蛋白酶体活性。蛋白质羰基含量的测定分析蛋白质氧化。应用免疫组织化学和激光共聚焦显微镜观察泛素在海马CA 1区神经元的分布。Western blotting检测不同缺血状态下细胞组分中蛋白聚集体、蛋白酶体、hsp 70和hsp 40的数量变化。结果如下：组织学检查显示，缺血后处理后，海马CA 1区存活神经元的百分比从5.21%±1.21%增加到55.32%±5.34%（P=0.0087）。Western blotting分析显示，缺血组再灌注12 h、24 h和48 h蛋白聚集体分别是假手术组的32.12±4.87、41.86±4.71和34.51±5.18倍。然而，缺血后处理在每个指定时间点显著减轻蛋白聚集体至2.84±0.97、13.72±2.13和14.37±2.42倍（P=0.000032、0.0000051和0.000082）。激光扫描共聚焦图像显示，缺血后处理组不能识别泛素标记的蛋白聚集体。进一步的研究表明，缺血后处理抑制了羰基衍生物的产生，提高了缺血和再灌注损伤的蛋白酶体活性，增加了伴侣蛋白hsp 70的表达，并维持了伴侣蛋白hsp 40的数量。结论：缺血后处理可显著挽救短暂缺血再灌注引起的CA 1区神经元死亡，这与抑制蛋白聚集的形成密切相关。
Objective: To investigate the effect of ischemic postconditioning on protein aggregation caused by transient ischemia and reperfusion and to clarify its underlying mechanism. Methods: Two-vessel-occluded transient global ischemia rat model was used. The rats in ischemic postconditioning group were subjected to three cycles of 30-s/30-s reperfusion/clamping after 15min of ischemia. Neuronal death in the CA1 region was observed by hematoxylin-eosin staining, and number of live neurons was assessed by cell counting under a light microscope. Succinyl-LLVY-AMC was used as substrate to assay proteasome activity in vitro. Protein carbonyl content was spectrophotometrically measured to analyze protein oxidization. Immunochemistry and laser scanning confocal microscopy were used to observe the distribution of ubiquitin in the CA1 neurons. Western blotting was used to analyze the quantitative alterations of protein aggregates, proteasome, hsp70 and hsp40 in cellular fractions under different ischemic conditions. Results: Histological examination showed that the percentage of live neurons in the CA1 region was elevated from 5.21%±1.21% to 55.32%±5.34% after administration of ischemic postconditioning (P=0.0087). Western blotting analysis showed that the protein aggregates in the ischemia group was 32.12±4.87, 41.86±4.71 and 34.51±5.18 times higher than that in the sham group at reperfusion 12h, 24h and 48h, respectively. However, protein aggregates were alleviated significantly by ischemic postconditioning to 2.84±0.97, 13.72±2.13 and 14.37±2.42 times at each indicated time point (P=0.000032, 0.0000051 and 0.0000082). Laser scanning confocal images showed ubiquitin labeled protein aggregates could not be discerned in the ischemic postconditioning group. Further study showed that ischemic postconditioning suppressed the production of carbonyl derivatives, elevated proteasome activity that was damaged by ischemia and reperfusion, increased the expression of chaperone hsp70, and maintained the quantity of chaperone hsp40. Conclusion: Ischemic postconditioning could rescue significantly neuronal death in the CA1 region caused by transient ischemia and reperfusion, which is closely associated with suppressing the formation of protein aggregation.
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影响因子：
3.3
作者：
Liu, CL;Ge, P;Hu, BR
通讯作者：
Hu, BR
影响因子：
3.9
作者：
Merker, K;Sitte, N;Grune, T
通讯作者：
Grune, T
影响因子：
8.3
作者：
Phillips, JB;Williams, AJ;Tortella, FC
通讯作者：
Tortella, FC
缺血后处理通过减弱蛋白质氧化来挽救局灶性缺血/再灌注引起的脑损伤
影响因子：
1.6
作者：
Li, Z. Y.;Liu, B.;Ge, P. F.
通讯作者：
Ge, P. F.
影响因子：
5.3
作者：
Hu, BR;Martone, ME;Liu, CL
通讯作者：
Liu, CL