Disruption of interleukin-1β autocrine signaling rescues complex I activity and improves ROS levels in immortalized epithelial cells with impaired cystic fibrosis transmembrane conductance regulator (CFTR) function.
Disruption of interleukin-1β autocrine signaling rescues complex I activity and improves ROS levels in immortalized epithelial cells with impaired cystic fibrosis transmembrane conductance regulator (CFTR) function.
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DOI：
10.1371/journal.pone.0099257
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Santa-Coloma TA
中科院分区：
文献类型：
--
作者：
Clauzure M;Valdivieso AG;Massip Copiz MM;Schulman G;Teiber ML;Santa-Coloma TA
囊性纤维化(CF)患者痰中细胞因子浓度升高，全身炎症状态。此外，培养的CF细胞过量产生多种细胞因子，包括IL-1β。我们之前已经证明，低剂量的IL-1β(∼30 pm)可以通过核转录因子-κB信号刺激T84结肠癌细胞中cftr的表达。然而，在较高剂量(2.5 ng/ml，∼150 pm)下，IL-1β抑制CFTRmRNA的表达。另一方面，通过差异显示，我们发现了两个在CF细胞中表达降低的基因，分别对应于线粒体蛋白CISD1和MTND4。最后一个是线粒体复合体I(MCX-I)活性的关键亚单位；相应地，我们后来在CF细胞中发现MCX-I活性降低。在此我们发现，在无血清培养条件下，IB3-1细胞在24小时内分泌的IL-1β为323±5 pg/ml，而S9细胞为127±3 pg/ml(校正后的IB3-1细胞)。外加IL-1shRNA(5 ng/ml)可降低S9(cftr-校正的ib3-1cf细胞)或Caco-2/pRSctrl细胞(shRNA对照细胞)的mCx-I活性，增加线粒体(mitoSOX探针)和细胞(DCFH-DA探针)ROS水平，与iB3-1或Caco-2/pRS26细胞(cftr特异性shRNA)的ROS水平相当。用IL-1β阻断抗体、IL-1受体拮抗剂、IKK抑制剂Ⅲ(NF-MAPK B通路)或SB203580(p38MAPK通路)处理IB3-1或Caco-2/pRS26细胞，均可恢复κ-I活性。此外，在IB3-1或Caco-2/pRS26细胞中，IL-1β阻断抗体、IKK抑制剂III或SB203580使线粒体ROS水平降低∼50%，细胞ROS水平接近基础值。AP-1抑制剂U0126(MEK1/2)或SP600125(JNK1/2/3)不起作用。结果提示，在这些细胞中，IL-1β通过自分泌效应起到了连接cftr与Mcx-I活性和ROS水平的桥梁作用。
Patients with cystic fibrosis (CF) have elevated concentration of cytokines in sputum and a general inflammatory condition. In addition, CF cells in culture produce diverse cytokines in excess, including IL-1β. We have previously shown that IL-1β, at low doses (∼30 pM), can stimulate the expression of CFTR in T84 colon carcinoma cells, through NF-κB signaling. However, at higher doses (>2.5 ng/ml, ∼150 pM), IL-1β inhibit CFTR mRNA expression. On the other hand, by using differential display, we found two genes with reduced expression in CF cells, corresponding to the mitochondrial proteins CISD1 and MTND4. The last is a key subunit for the activity of mitochondrial Complex I (mCx-I); accordingly, we later found a reduced mCx-I activity in CF cells. Here we found that IB3-1 cells (CF cells), cultured in serum-free media, secrete 323±5 pg/ml of IL-1β in 24 h vs 127±3 pg/ml for S9 cells (CFTR-corrected IB3-1 cells). Externally added IL-1β (5 ng/ml) reduces the mCx-I activity and increases the mitochondrial (MitoSOX probe) and cellular (DCFH-DA probe) ROS levels of S9 (CFTR-corrected IB3-1 CF cells) or Caco-2/pRSctrl cells (shRNA control cells) to values comparable to those of IB3-1 or Caco-2/pRS26 cells (shRNA specific for CFTR). Treatments of IB3-1 or Caco-2/pRS26 cells with either IL-1β blocking antibody, IL-1 receptor antagonist, IKK inhibitor III (NF-κB pathway) or SB203580 (p38 MAPK pathway), restored the mCx-I activity. In addition, in IB3-1 or Caco-2/pRS26 cells, IL-1β blocking antibody, IKK inhibitor III or SB203580 reduced the mitochondrial ROS levels by ∼50% and the cellular ROS levels near to basal values. The AP-1 inhibitors U0126 (MEK1/2) or SP600125 (JNK1/2/3 inhibitor) had no effects. The results suggest that in these cells IL-1β, through an autocrine effect, acts as a bridge connecting the CFTR with the mCx-I activity and the ROS levels.
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影响因子：
4.2
作者：
Alekseeva L;Huet D;Féménia F;Mouyna I;Abdelouahab M;Cagna A;Guerrier D;Tichanné-Seltzer V;Baeza-Squiban A;Chermette R;Latgé JP;Berkova N
通讯作者：
Berkova N
DOI：
10.1073/pnas.251194298
发表时间：
2001-11-20
影响因子：
11.1
作者：
Bennett, BL;Sasaki, DT;Anderson, DW
通讯作者：
Anderson, DW
DOI：
10.1164/rccm.200207-765oc
发表时间：
2004-03-01
影响因子：
24.7
作者：
Becker, MN;Sauer, MS;Randell, SH
通讯作者：
Randell, SH
影响因子：
64.8
作者：
EGAN, M;FLOTTE, T;GUGGINO, WB
通讯作者：
GUGGINO, WB
影响因子：
4.8
作者：
Cafferata, EGA;Guerrico, AMG;Santa-Coloma, TA
通讯作者：
Santa-Coloma, TA