Interleukin-11-induced heat shock protein 25 confers intestinal epithelial-specific cytoprotection from oxidant stress
Interleukin-11-induced heat shock protein 25 confers intestinal epithelial-specific cytoprotection from oxidant stress
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DOI：
10.1016/s0016-5085(03)00282-8
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发表时间：
2003-05-01
期刊：
影响因子：
29.4
通讯作者：
Chang, EB
中科院分区：
文献类型：
--
作者：
Ropeleski, MJ;Tang, J;Chang, EB
背景与目的：白细胞介素-11（IL-11）在肠上皮损伤中的细胞保护机制尚不清楚。IL-11在氧化应激期间保护屏障完整性，氧化应激是多种类型的肠损伤（包括缺血和免疫介导的炎症）的共同终点。由于热休克蛋白（hsp）在肠上皮细胞中具有细胞保护作用，我们假设IL-11赋予的细胞保护作用是由诱导型hsp介导的。方法：IL-11受体（IL-11 R）的激活使用磷酸化特异性抗体STAT 3。通过免疫印迹法测定IEC-18隐窝和年轻成年小鼠结肠上皮细胞中IL-11对hsp 72和hsp 25的诱导。通过Cr-51释放测定上皮细胞对一氯胺氧化损伤的抗性。通过电穿孔和潮霉素B筛选获得稳定的hsp反义IEC-18细胞克隆。通过Triton X-100不溶性组分、2-D等电聚焦凝胶和共聚焦显微镜分析IL-11对hsp 25分布的影响。结果：在所研究的所有细胞中检测到IL-11 R信号。IL-11以肠上皮特异性方式诱导hsp 25，其在一氯胺存在下显著保持细胞活力。这种作用在稳定表达hsp 25反义的肠上皮细胞中被显著逆转。IL-11诱导hsp 25转移到含有细胞骨架元素的Triton X-100不溶性组分，这与hsp 25磷酸化的改变无关。通过共聚焦显微镜观察发现，热休克蛋白25与F-肌动蛋白共定位的增加并不能阻止这种变化。结论：IL-11诱导hsp 25表达。赋予上皮特异性细胞保护作用，其不依赖于hsp 25与F-肌动蛋白的磷酸化依赖性共定位，从而有助于IL-11在肠上皮损伤模型中的保护作用。
Background & Aims: The mechanisms of interleukin-11 (IL-11) cytoprotection in intestinal epithelial injury are largely unknown. IL-11 protects barrier integrity during oxidant stress, a common endpoint of numerous types of intestinal injury including ischemia and immune-mediated inflammation. Because heat shock proteins (hsp) are cytoprotective in intestinal epithelia, we hypothesized that IL-11- conferred cytoprotection is mediated by inducible hsps. Methods: IL-11 receptor (IL-11R) activation was determined using phospho-specific antibodies to STAT3. IL-11 induction of hsp72 and hsp25 was determined by immunoblot in IEC-18 crypt and young adult mouse colon colonic epithelial cells. Epithelial resistance to oxidant injury by monochloramine was determined by Cr-51 release. Stable hsp anti-sense IEC-18 cell clones were obtained by electroporation and hygromycin B selection. The IL-11 effect on hsp25 distribution was characterized by analysis of Triton X-100 insoluble fractions, 2-D isoelectric focusing gels, and confocal microscopy. Results: IL-11R signaling was detected in all cells under study. IL-11 induces hsp25 in an intestinal epithelial-specific manner that significantly preserves cellular viability in the presence of monochloramine. This effect was significantly reversed in intestinal epithelia stably expressing anti-sense to hsp25. IL-11 induced a shift of hsp25 to Triton X-100 insoluble fractions containing cytoskeletal elements, which was not associated with altered hsp25 phosphorylation. The shift was not paralleled by increased hsp25 co-localization with F-actin by confocal microscopy. Conclusions: The induction of hsp25 by IL-11. confers epithelial-specific cytoprotection that is independent of phosphorylation-dependent co-localization of hsp25 to F-actin, thereby contributing to the protective effects of IL-11 in models of intestinal epithelial injury.