Follistatin-Like 1 Promotes Bleomycin-Induced Pulmonary Fibrosis through the Transforming Growth Factor Beta 1/Mitogen-Activated Protein Kinase Signaling Pathway.
Follistatin-Like 1 Promotes Bleomycin-Induced Pulmonary Fibrosis through the Transforming Growth Factor Beta 1/Mitogen-Activated Protein Kinase Signaling Pathway.
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Follistatin-Like 1 通过转化生长因子 Beta 1/丝裂原激活蛋白激酶信号通路促进博来霉素诱导的肺纤维化
DOI：
10.4103/0366-6999.238151
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发表时间：
2018-08-20
影响因子：
6.1
通讯作者：
Wang C
中科院分区：
文献类型：
--
作者：
Jin YK;Li XH;Wang W;Liu J;Zhang W;Fang YS;Zhang ZF;Dai HP;Ning W;Wang C
卵泡抑素样蛋白1（FSTL 1）是一种新的促纤维化因子，通过转化生长因子β 1（TGF-β1）/Smad信号通路诱导肺纤维化。其通过非Smad信号途径如丝裂原活化蛋白激酶（MAPK）途径对PF的影响知之甚少。因此，本研究旨在探讨FSTL 1通过MAPK信号通路在PF中的作用及其在肺纤维化发生中的机制。在Fstl 1 +/−和野生型（WT）C57 BL/6小鼠中用博来霉素诱导PF。14 d后处死小鼠，取肺组织行苏木精-伊红染色，测定肺组织羟脯氨酸含量以确定PF，采用定量聚合酶链反应和Western blotting检测FSTL 1 mRNA和蛋白水平及MAPK磷酸化水平的变化。通过Western印迹和细胞免疫荧光检测Fstl 1缺陷对成纤维细胞分化的影响。在用重组人FSTL 1蛋白处理的Fstl 1 +/-和WT成纤维细胞中通过蛋白质印迹法测量MAPK信号传导激活。我们用细胞外信号调节激酶（ERK）、p38和Jun N-末端激酶（JNK）信号抑制剂预处理小鼠肺成纤维细胞，并通过Western印迹、3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四氮唑分析和transwell测定分析其分化、增殖、迁移和侵袭。学生t检验用于比较两组之间的差异。Fstl 1缺陷减弱了博莱霉素诱导的肺纤维化组织损伤后14天ERK、p38和JNK信号的磷酸化（0.67 ± 0.05 vs.1.22 ± 0.03，t = 14.92，P = 0.0001; 0.41 ± 0.01 vs.1.15 ± 0.07; t = 11.19; P = 0.0004;和0.41 ± 0.01 vs. 1.07 ± 0.07，t = 8.92，P = 0.0009;分别），与同一时间WT肺和原代肺成纤维细胞相比（0.82 ± 0.01 vs. 1.01 ± 0.04，t = 4.06，P = 0.0150; 1.04 ± 0.03 vs. 1.24 ± 0.03，t = 4.44，P = 0.0100; 0.76 ± 0.05 vs. 0.99 ± 0.05，t = 4.48，P = 0.0100;与TGF-β1刺激的WT组相比。肺成纤维细胞中重组人FSTL 1蛋白增强TGF-β1介导的ERK磷酸化（1.19 ± 0.08 vs. 0.55 ± 0.04，t = 6.99，P = 0.0020），p38（1.18 ± 0.04 vs.0.66 ± 0.03，t = 11.20，P = 0.0020），JNK（1.11 ± 0.01 vs.0.84 ± 0.04，t = 6.53，P = 0.0030）。Fstl 1缺陷的成纤维细胞显示α-平滑肌肌动蛋白（α-SMA）表达减少（0.70 ± 0.06 vs.1.28 ± 0.11，t = 4.65，P = 0.0035; 1.40 ± 0.05 vs.1.76 ± 0.02，t = 6.31，P = 0.0007;与TGF-β1处理的WT组相比）。与对照组相比，p38抑制剂预处理可显著抑制TGF-β1/FSTL 1介导的α-SMA表达（0.73 ± 0.01 vs.1.13 ± 0.10，t = 3.92，P = 0.0078）和JNK（0.78 ± 0.03 vs.1.08 ± 0.06，t = 4.40，P = 0.0046）信号。小鼠肺成纤维细胞（MLgs）的增殖在p38抑制剂处理后显著降低（0.30 ± 0.01 vs. 0.46 ± 0.03，t = 4.64，P = 0.0009），JNK（0.30 ± 0.01 vs. 0.49 ± 0.01，t = 12.84，P = 0.0001）和Smad 2/3（0.18 ± 0.02 vs. 0.46 ± 0.02，t = 12.69，P = 0.0001）信号转导。在p38抑制剂预处理的培养液中，MLgs的迁移和侵袭能力明显降低（移行细胞为70.17 ± 3.28 vs.116.30 ± 7.11，t = 5.89，P = 0.0042; 19.87 ± 0.84 vs.32.70 ± 0.95，t = 10.14，P = 0.0005），JNK（72.30 ± 3.85vs.116.30 ± 7.11，t = 5.44，P = 0.0056）; 18.03 ± 0.94 vs.32.70 ± 0.95，对于侵袭性细胞，t = 11.00，P = 0.0004）和Smad 2/3（64.76 ± 1.41 vs. 116.30 ± 7.11，t = 7.11，P = 0.0021）; 18.03 ± 0.94 vs.32.70 ± 0.95，对于侵袭性细胞，t = 13.29，P = 0.0002）信号传导与二甲基亚砜组中的相应条件相比。FSTL 1通过p38和JNK信号通路影响肺成纤维细胞的分化、增殖、迁移和侵袭，从而影响肺纤维化的发生发展。
Follistatin-like 1 (FSTL1) is a novel profibrogenic factor that induces pulmonary fibrosis (PF) through the transforming growth factor-beta 1 (TGF-β1)/Smad signaling. Little is known about its effects on PF through the non-Smad signaling, like the mitogen-activated protein kinase (MAPK) pathway. Therefore, this study aimed to investigate the role of FSTL1 in PF through the MAPK signaling pathway and its mechanisms in lung fibrogenesis. PF was induced in Fstl1+/− and wild-type (WT) C57BL/6 mice with bleomycin. After 14 days, the mice were sacrificed, and lung tissues were stained with hematoxylin and eosin; the hydroxyproline content was measured to confirm PF. The mRNA and protein level of FSTL1 and the change of MAPK phosphorylation were measured by quantitative polymerase chain reaction and Western blotting. The effect of Fstl1 deficiency on fibroblasts differentiation was measured by Western blotting and cell immunofluorescence. MAPK signaling activation was measured by Western blotting in Fstl1+/− and WT fibroblasts treated with recombinant human FSTL1 protein. We pretreated mouse lung fibroblast cells with inhibitors of the extracellular signal-regulated kinase (ERK), p38, and Jun N-terminal kinase (JNK) signaling and analyzed their differentiation, proliferation, migration, and invasion by Western blotting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis, and transwell assays. The Student's t-test was used to compare the differences between two groups. Fstl1 deficiency attenuated phosphorylation of the ERK, p38, and JNK signaling in bleomycin-induced fibrotic lung tissue 14 days after injury (0.67 ± 0.05 vs. 1.22 ± 0.03, t = 14.92, P = 0.0001; 0.41 ± 0.01 vs. 1.15 ± 0.07; t = 11.19; P = 0.0004; and 0.41 ± 0.01 vs. 1.07 ± 0.07, t = 8.92, P = 0.0009; respectively), compared with WT lungs at the same time and in primary lung fibroblasts (0.82 ± 0.01 vs. 1.01 ± 0.04, t = 4.06, P = 0.0150; 1.04 ± 0.03 vs. 1.24 ± 0.03, t = 4.44, P = 0.0100; and 0.76 ± 0.05 vs. 0.99 ± 0.05, t = 4.48, P = 0.0100; respectively), compared with TGF-β1-stimulated WT group. Recombinant human FSTL1 protein in lung fibroblasts enhanced TGF-β1-mediated phosphorylation of the ERK (1.19 ± 0.08 vs. 0.55 ± 0.04, t = 6.99, P = 0.0020), p38 (1.18 ± 0.04 vs. 0.66 ± 0.03, t = 11.20, P = 0.0020), and JNK (1.11 ± 0.01 vs. 0.84 ± 0.04, t = 6.53, P = 0.0030), compared with the TGF-β1-stimulated WT group. Fstl1-deficient fibroblasts showed reduced alpha-smooth muscle actin (α-SMA) expression (0.70 ± 0.06 vs. 1.28 ± 0.11, t = 4.65, P = 0.0035, compared with the untreated WT group; 1.40 ± 0.05 vs. 1.76 ± 0.02, t = 6.31, P = 0.0007; compared with the TGF-β1-treated WT group). Compared with the corresponding condition in the control group, the TGF-β1/FSTL1-mediated α-SMA expression was significantly suppressed by pretreatment with an inhibitor of p38 (0.73 ± 0.01 vs. 1.13 ± 0.10, t = 3.92, P = 0.0078) and JNK (0.78 ± 0.03 vs. 1.08 ± 0.06, t = 4.40, P = 0.0046) signaling. The proliferation of mouse lung fibroblast cells (MLgs) significantly decreased after treatment of an inhibitor of p38 (0.30 ± 0.01 vs. 0.46 ± 0.03, t = 4.64, P = 0.0009), JNK (0.30 ± 0.01 vs. 0.49 ± 0.01, t = 12.84, P = 0.0001), and Smad2/3 (0.18 ± 0.02 vs. 0.46 ± 0.02, t = 12.69, P = 0.0001) signaling compared with the dimethylsulfoxide group. The migration and invasion cells of MLgs significantly decreased in medium pretreated with an inhibitor of p38 (70.17 ± 3.28 vs. 116.30 ± 7.11, t = 5.89, P = 0.0042 for the migratory cells; 19.87 ± 0.84 vs. 32.70 ± 0.95, t = 10.14, P = 0.0005 for the invasive cells), JNK (72.30 ± 3.85 vs. 116.30 ± 7.11, t = 5.44, P = 0.0056 for the migratory cells; 18.03 ± 0.94 vs. 32.70 ± 0.95, t = 11.00, P = 0.0004 for the invasive cells), and Smad2/3 (64.76 ± 1.41 vs. 116.30 ± 7.11, t = 7.11, P = 0.0021 for the migratory cells; 18.03 ± 0.94 vs. 32.70 ± 0.95, t = 13.29, P = 0.0002 for the invasive cells) signaling compared with the corresponding condition in the dimethylsulfoxide group. FSTL1 affects lung fibroblast differentiation, proliferation, migration, and invasion through p38 and JNK signaling, and in this way, it might influence the development of PF.
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阻断卵泡抑素样 1 可减轻博莱霉素诱导的小鼠肺纤维化
DOI：
10.1084/jem.20121878
发表时间：
2015-02-09
期刊：
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影响因子：
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影响因子：
10.6
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van der Velden JL;Ye Y;Nolin JD;Hoffman SM;Chapman DG;Lahue KG;Abdalla S;Chen P;Liu Y;Bennett B;Khalil N;Sutherland D;Smith W;Horan G;Assaf M;Horowitz Z;Chopra R;Stevens RM;Palmisano M;Janssen-Heininger YM;Schafer PH
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