Enhanced Hsp70 expression protects against acute lung injury by modulating apoptotic pathways.
Enhanced Hsp70 expression protects against acute lung injury by modulating apoptotic pathways.
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增强的HSP70表达通过调节凋亡途径来预防急性肺损伤。
DOI：
10.1371/journal.pone.0026956
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Weiss YG
中科院分区：
文献类型：
--
作者：
Aschkenasy G;Bromberg Z;Raj N;Deutschman CS;Weiss YG
急性呼吸窘迫综合征（ARDS）是一种高度致命的炎症性肺部疾病。细胞凋亡在其发病机制中起关键作用。我们发现，腺病毒表达的70 kDa热休克蛋白70（AdHSP）保护脓毒症引起的肺损伤。在这项研究中，我们测试的假设，AdHSP减弱脓毒症引起的肺损伤的细胞凋亡。通过盲肠结扎和双重穿刺（2CLP）在大鼠中诱导脓毒症。在2CLP PBS时，将AdHSP或AdGFP（表达绿色荧光蛋白的腺病毒载体）注射到脓毒症大鼠的气管中。48小时后，分离肺。将一侧肺固定用于TUNEL染色和免疫组织化学。另一组匀浆分离胞浆和核蛋白。在这些提取物中进行免疫印迹、凝胶过滤和免疫共沉淀。在单独的实验中，将MLE-12细胞与培养基、AdHSP或AdGFP孵育。用TNFα刺激细胞。分离胞浆和核蛋白。对这些进行免疫印迹、免疫共沉淀和半胱天冬酶-3活性测定。TUNEL法显示AdHSP可减少肺泡细胞凋亡。免疫组化检测caspase 3丰度证实了这一点。在从脓毒症动物中分离的肺中，免疫印迹、免疫共沉淀和凝胶过滤研究显示含有半胱天冬酶3、8和9的细胞质复合物增加。AdHSP破坏了这些复合物。我们认为热休克蛋白70通过与半胱天冬酶的相互作用损害细胞凋亡途径。大复合物的破坏导致较低分子量复合物的稳定，从而减少核半胱天冬酶-3。预防肺损伤中的细胞凋亡可以保护肺泡细胞并有助于恢复。
The Acute respiratory distress syndrome (ARDS) is a highly lethal inflammatory lung disorder. Apoptosis plays a key role in its pathogenesis. We showed that an adenovirus expressing the 70 kDa heat shock protein Hsp70 (AdHSP) protected against sepsis-induced lung injury. In this study we tested the hypothesis that AdHSP attenuates apoptosis in sepsis-induced lung injury. Sepsis was induced in rats via cecal ligation and double puncture (2CLP). At the time of 2CLP PBS, AdHSP or AdGFP (an adenoviral vector expressing green fluorescent protein) were injected into the tracheas of septic rats. 48 hours later, lungs were isolated. One lung was fixed for TUNEL staining and immunohistochemistry. The other was homogenized to isolate cytosolic and nuclear protein. Immunoblotting, gel filtration and co-immunoprecipitation were performed in these extracts. In separate experiments MLE-12 cells were incubated with medium, AdHSP or AdGFP. Cells were stimulated with TNFα. Cytosolic and nuclear proteins were isolated. These were subjected to immunoblotting, co- immunoprecipitation and a caspase-3 activity assay. TUNEL assay demonstrated that AdHSP reduced alveolar cell apoptosis. This was confirmed by immunohistochemical detection of caspase 3 abundance. In lung isolated from septic animals, immunoblotting, co-immunoprecipitation and gel filtration studies revealed an increase in cytoplasmic complexes containing caspases 3, 8 and 9. AdHSP disrupted these complexes. We propose that Hsp70 impairs apoptotic cellular pathways via interactions with caspases. Disruption of large complexes resulted in stabilization of lower molecular weight complexes, thereby, reducing nuclear caspase-3. Prevention of apoptosis in lung injury may preserve alveolar cells and aid in recovery.
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影响因子：
4.8
作者：
Delettre, C;Yuste, VJ;Susin, SA
通讯作者：
Susin, SA
影响因子：
11.4
作者：
Jäättelä, M;Wissing, D;Egeblad, M
通讯作者：
Egeblad, M
影响因子：
6.1
作者：
Oyake, Jinko;Otaka, Michiro;Watanabe, Sumio
通讯作者：
Watanabe, Sumio
DOI：
10.1083/jcb.119.3.493
发表时间：
1992-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Gavrieli Y;Sherman Y;Ben-Sasson SA
通讯作者：
Ben-Sasson SA
影响因子：
3.6
作者：
Diamant, S;Rosenthal, D;Goloubinoff, P
通讯作者：
Goloubinoff, P