ER Stress via CHOP Pathway is Involved in FK506-Induced Apoptosis in Rat Fibroblasts
ER Stress via CHOP Pathway is Involved in FK506-Induced Apoptosis in Rat Fibroblasts
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通过 CHOP 途径的 ER 应激参与 FK506 诱导的大鼠成纤维细胞凋亡
DOI：
10.1159/000447894
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发表时间：
2016-01-01
影响因子：
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通讯作者：
Cao, Xiaojian
中科院分区：
文献类型：
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作者：
Tang, Jian;Ge, Yingbin;Cao, Xiaojian
背景/目的：增生性瘢痕(HS)的形成是由于成纤维细胞凋亡减少和增殖增加所致。因此，成纤维细胞的凋亡是开发HS新的治疗策略的关键靶点。已有报道表明，FK506可减轻体内瘢痕形成，并可诱导内质网应激(ER应激)。然而，FK506在内质网应激介导的成纤维细胞凋亡中的作用尚不清楚。方法：以大鼠皮肤成纤维细胞为研究对象。用细胞计数试剂盒检测细胞活性。Annexin V/碘化丙啶双染色法检测细胞凋亡。基因沉默是使用小干扰RNA(SiRNAs)或通过慢病毒感染来实现的。免疫印迹法检测细胞凋亡相关蛋白的表达。用免疫共沉淀法研究蛋白质之间的相互作用。结果：FK506可显著降低成纤维细胞的存活率，诱导成纤维细胞的凋亡。有趣的是，FK506处理后，内质网应激也被激活。我们进一步证明，FK506诱导的细胞凋亡是由内质网应激通过激活CHOP介导的，用TUDCA抑制ER应激或沉默CHOP的表达后，细胞凋亡率减少。免疫共沉淀结果表明，FK506处理可诱导FKBP12.6从RyR2解离并从胞膜移位至胞浆，从而促进内质网应激诱导的细胞凋亡。结论：内质网应激通过CHOP信号通路介导FK506诱导成纤维细胞凋亡。(C)2016年作者(S)，S.Karger AG，巴塞尔出版
Background/Aims: Hypertrophic scars (HS) formation results from reduced apoptosis and increased proliferation of fibroblasts. Therefore, apoptosis of fibroblasts is a key target for the development of novel therapeutic strategies for HS. Previous reports demonstrated that FK506 could attenuate scar formation in vivo and FK506 could also induce endoplasmic reticulum stress (ER stress). However, the effects of FK506 on ER stress-mediated apoptosis in fibroblasts remain unclear. Methods: Rat skin fibroblasts were used in the study. Cell viability was examined using cell counting Kit-8. Apoptosis was detected by Annexin V/Propidium Iodide Double Staining. Gene silencing was performed using Small Interfering RNAs (siRNAs) or via lentiviral infection. The expression of apoptosis-related proteins was determined via Western blot. Interaction between proteins was explored by co-immunoprecipitation. Results: FK506 significantly reduced cell viability and induced apoptosis in fibroblasts. Interestingly, ER stress was also activated after FK506 treatment. We further demonstrated that FK506-induced apoptosis was mediated by ER stress via activating CHOP, evidenced by decreased apoptosis after inhibition of ER stress using TUDCA or silencing expression of CHOP. Furthermore, Co-immunoprecipitation results indicated that treatment of FK506 induced disassociation of FKBP12.6 from RyR2 and its translocation from ER membrane to cytosol, consequently promoting ER stress-mediated apoptosis. Conclusion: FK506-induced fibroblasts apoptosis was mediated by ER stress via CHOP signaling pathway. (C) 2016 The Author(s) Published by S. Karger AG, Basel