Proteomic Profiling of Radiation-Induced Skin Fibrosis in Rats: Targeting the Ubiquitin-Proteasome System
Proteomic Profiling of Radiation-Induced Skin Fibrosis in Rats: Targeting the Ubiquitin-Proteasome System
复制标题
大鼠辐射诱导皮肤纤维化的蛋白质组学分析：针对泛素-蛋白酶体系统
DOI：
10.1016/j.ijrobp.2016.01.021
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发表时间：
2016-06-01
影响因子：
7
通讯作者：
Zhang, Shuyu
中科院分区：
文献类型：
--
作者：
Wang, Wenjie;Luo, Judong;Zhang, Shuyu
目的：研究辐射引起的皮肤纤维化发病机制中的分子变化。方法和材料：用电子束对大鼠皮肤进行30或45Gy的照射。通过无标记蛋白质定量来定量纤维化大鼠皮肤和邻近正常组织中的蛋白质表达。通过X射线照射处理人皮肤细胞HaCaT和WS-1，并用荧光探针测定蛋白酶体活性。通过蛋白质印迹和免疫荧光测量蛋白酶体抑制剂对转化生长因子β (TGF-B) 信号传导的影响。通过皮肤损伤量表评估硼替佐米对大鼠皮肤伤口愈合的功效。结果：我们发现辐射引起大鼠和人类皮肤的表皮和真皮增生。在受辐射的纤维化皮肤中鉴定出 196 种优先表达的蛋白质和 80 种独特的蛋白质。通过生物信息学分析，泛素-蛋白酶体途径显示出显着的倍数变化，并进行了更详细的研究。体外实验表明，辐射导致人体皮肤细胞中蛋白酶体的活性下降。蛋白酶体抑制剂硼替佐米抑制促纤维化 TGF-β 下游信号传导，但不抑制 HaCaT 和 WS-1 细胞中辐射刺激的 TGF-β 分泌。此外，硼替佐米可改善辐射引起的皮肤损伤并减轻表皮增生。结论：我们的研究结果说明了辐射引起的皮肤纤维化过程中的分子变化，并表明针对泛素蛋白酶体系统将是一种有效的对策。 (C) 2016 Elsevier Inc. 保留所有权利。
Purpose: To investigate the molecular changes underlying the pathogenesis of radiation-induced skin fibrosis.Methods and Materials: Rat skin was irradiated to 30 or 45 Gy with an electron beam. Protein expression in fibrotic rat skin and adjacent normal tissues was quantified by label-free protein quantitation. Human skin cells HaCaT and WS-1 were treated by x-ray irradiation, and the proteasome activity was determined with a fluorescent probe. The effect of proteasome inhibitors on Transforming growth factor Beta (TGF-B) signaling was measured by Western blot and immunofluorescence. The efficacy of bortezomib in wound healing of rat skin was assessed by the skin injury scale.Results: We found that irradiation induced epidermal and dermal hyperplasia in rat and human skin. One hundred ninety-six preferentially expressed and 80 unique proteins in the irradiated fibrotic skin were identified. Through bioinformatic analysis, the ubiquitin-proteasome pathway showed a significant fold change and was investigated in greater detail. In vitro experiments demonstrated that irradiation resulted in a decline in the activity of the proteasome in human skin cells. The proteasome inhibitor bortezomib suppressed profibrotic TGF-beta downstream signaling but not TGF-beta secretion stimulated by irradiation in HaCaT and WS-1 cells. Moreover, bortezomib ameliorated radiation-induced skin injury and attenuated epidermal hyperplasia.Conclusion: Our findings illustrate the molecular changes during radiation-induced skin fibrosis and suggest that targeting the ubiquitin-proteasome system would be an effective countermeasure. (C) 2016 Elsevier Inc. All rights reserved.