Chronic exposure of interleukin-13 suppress the induction of matrix metalloproteinase-1 by tumour necrosis factor in normal and scleroderma dermal fibroblasts through protein kinase B/Akt
Chronic exposure of interleukin-13 suppress the induction of matrix metalloproteinase-1 by tumour necrosis factor in normal and scleroderma dermal fibroblasts through protein kinase B/Akt
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DOI：
10.1111/cei.13045
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发表时间：
2018-01-01
影响因子：
4.6
通讯作者：
Hasty, K. A.
中科院分区：
文献类型：
--
作者：
Lobbins, M. L. Brown;Shivakumar, B. R.;Hasty, K. A.
从硬皮病患者获取的外周血单个核细胞表达升高水平的白细胞介素（IL）-13。此外，来自受累硬皮病皮肤成纤维细胞的基质金属蛋白酶 -1（MMP -1）的表达对肿瘤坏死因子（TNF）-α刺激无反应。为了阐明所涉及的机制，我们检测了IL -13对正常和硬皮病人类皮肤成纤维细胞系中TNF -α诱导的MMP -1表达的影响，并研究了丝氨酸/苏氨酸激酶B/蛋白激酶B（Akt）在这种反应中的作用。皮肤成纤维细胞系在不同浓度IL -13存在的情况下用TNF -α刺激。使用蛋白质印迹分析对总Akt和磷酸化Akt（pAkt）进行定量。成纤维细胞在有或无Akt抑制剂VIII的情况下用IL -13处理，然后用TNF -α刺激。通过实时聚合酶链反应（PCR）和酶联免疫吸附测定（ELISA）分析MMP -1的表达。使用方差分析（ANOVA）或学生t检验进行统计分析。在TNF -α刺激下，正常皮肤成纤维细胞比系统性硬化症（SSc）成纤维细胞分泌更多的MMP -1。当成纤维细胞与IL -13和TNF -α共同孵育时，MMP -1的这种增加消失。IL -13诱导皮肤成纤维细胞中pAkt水平显著升高，而Akt抑制剂VIII逆转了IL -13对培养的成纤维细胞对TNF -α反应的抑制作用，增加了它们的MMP -1表达。我们表明IL -13抑制TNF -α刺激的正常和硬皮病皮肤成纤维细胞中的MMP -1。Akt抑制剂VIII能够逆转IL -13对MMP -1表达和蛋白质合成的抑制作用。我们的数据表明IL -13通过Akt介导的途径调节对TNF -α的MMP -1表达，并可能在硬皮病等纤维化疾病中起作用。
Peripheral blood mononuclear cells taken from patients with scleroderma express increased levels of interleukin (IL)-13. Moreover, the expression of matrix metalloproteinase-1 (MMP-1) from involved scleroderma skin fibroblasts is refractory to stimulation by tumour necrosis factor (TNF)-. To elucidate the mechanism(s) involved, we examined the effect of IL-13 on TNF--induced MMP-1 expression in normal and scleroderma human dermal fibroblast lines and studied the involvement of serine/threonine kinase B/protein kinase B (Akt) in this response. Dermal fibroblast lines were stimulated with TNF- in the presence of varying concentrations of IL-13. Total Akt and pAkt were quantitated using Western blot analyses. Fibroblasts were treated with or without Akt inhibitor VIII in the presence of IL-13 followed by TNF- stimulation. MMP-1 expression was analysed by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed using analysis of variance (anova) or Student's t-test. Upon TNF- stimulation, normal dermal fibroblasts secrete more MMP-1 than systemic sclerosis (SSc) fibroblasts. This increase in MMP-1 is lost when fibroblasts are co-incubated with IL-13 and TNF-. IL-13 induced a significant increase in levels of pAkt in dermal fibroblasts, while Akt inhibitor VIII reversed the suppressive effects of IL-13 on the response of cultured fibroblasts to TNF-, increasing their expression of MMP-1. We show that IL-13 suppresses MMP-1 in TNF--stimulated normal and scleroderma dermal fibroblast. Akt inhibitor VIII is able to reverse the suppressive effect of IL-13 on MMP-1 expression and protein synthesis. Our data suggest that IL-13 regulates MMP-1 expression in response to TNF- through an Akt-mediated pathway and may play a role in fibrotic diseases such as scleroderma.