DIDS prevents ischemic membrane degradation in cultured hippocampal neurons by inhibiting matrix metalloproteinase release.
DIDS prevents ischemic membrane degradation in cultured hippocampal neurons by inhibiting matrix metalloproteinase release.
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DOI：
10.1371/journal.pone.0043995
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Haddad GG
中科院分区：
文献类型：
--
作者：
Pamenter ME;Ryu J;Hua ST;Perkins GA;Mendiola VL;Gu XQ;Ellisman MH;Haddad GG
在中风期间，梗塞核心中的细胞表现出其渗透性屏障的快速失效，其释放对附近组织（半影）有害的离子和炎性分子。质膜降解是半影扩散的关键，并由基质金属蛋白酶（MMPs）介导，MMPs在应激时通过囊泡胞吐作用释放到细胞外液中。DIDS（4，4 ′-二异硫氰基芪-2，2 ′-二磺酸）在体外缺血半暗带模拟物（缺血溶液：IS）激发的神经元中保持膜完整性，我们询问这种作用是否通过抑制MMP活性介导。在培养的小鼠海马神经元中，用IS攻击，细胞内proMMP-2和-9表达增加4-10倍，细胞外潜伏和活性MMP亚型表达增加2-22倍。MMP介导的细胞外明胶分解活性增加了20-50倍，导致32.1±4.5%的细胞从基质中脱离，并引起广泛的质膜降解（>60%的细胞摄取活体染料，>60%的质膜破碎或起泡）。DIDS消除了神经元中的细胞脱离和膜降解以及病理诱导的潜伏和活性MMPs的细胞外表达。DIDS类似地抑制细胞外MMP表达和由促凋亡剂星形孢菌素或一般蛋白酶激动剂4-氨基苯汞乙酸（APMA）诱导的细胞脱离。相反，DIDS治疗不损害应激诱导的细胞内proMMP的产生，也不损害proMMP-2的细胞内裂解为活性形式，这表明DIDS干扰MMP的囊泡挤出，而不是直接抑制蛋白酶的表达或活化。在支持这一假设，拮抗剂的V型囊泡ATP酶也抑制细胞外MMP的表达，以类似的程度作为DIDS。此外，在一个蛋白酶非依赖性模型的囊泡胞吐，DIDS阻止刺激诱发的释放血管性血友病因子从人脐静脉内皮细胞。我们的结论是，DIDS抑制MMP胞吐，并通过这种机制保持神经元膜的完整性，在病理应激。
During stroke, cells in the infarct core exhibit rapid failure of their permeability barriers, which releases ions and inflammatory molecules that are deleterious to nearby tissue (the penumbra). Plasma membrane degradation is key to penumbral spread and is mediated by matrix metalloproteinases (MMPs), which are released via vesicular exocytosis into the extracellular fluid in response to stress. DIDS (4,4′-diisothiocyanatostilbene-2,2′-disulphonic acid) preserves membrane integrity in neurons challenged with an in vitro ischemic penumbral mimic (ischemic solution: IS) and we asked whether this action was mediated via inhibition of MMP activity. In cultured murine hippocampal neurons challenged with IS, intracellular proMMP-2 and -9 expression increased 4–10 fold and extracellular latent and active MMP isoform expression increased 2–22 fold. MMP-mediated extracellular gelatinolytic activity increased ∼20–50 fold, causing detachment of 32.1±4.5% of cells from the matrix and extensive plasma membrane degradation (>60% of cells took up vital dyes and >60% of plasma membranes were fragmented or blebbed). DIDS abolished cellular detachment and membrane degradation in neurons and the pathology-induced extracellular expression of latent and active MMPs. DIDS similarly inhibited extracellular MMP expression and cellular detachment induced by the pro-apoptotic agent staurosporine or the general proteinase agonist 4-aminophenylmercuric acetate (APMA). Conversely, DIDS-treatment did not impair stress-induced intracellular proMMP production, nor the intracellular cleavage of proMMP-2 to the active form, suggesting DIDS interferes with the vesicular extrusion of MMPs rather than directly inhibiting proteinase expression or activation. In support of this hypothesis, an antagonist of the V-type vesicular ATPase also inhibited extracellular MMP expression to a similar degree as DIDS. In addition, in a proteinase-independent model of vesicular exocytosis, DIDS prevented stimulus-evoked release of von Willebrand Factor from human umbilical vein endothelial cells. We conclude that DIDS inhibits MMP exocytosis and through this mechanism preserves neuronal membrane integrity during pathological stress.
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影响因子：
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作者：
Liu, An-Heng;Cao, Ya-Nan;Wang, Xiao-Ming
通讯作者：
Wang, Xiao-Ming
影响因子：
--
作者：
Leppert, D;Lindberg, RLP;Leib, SL
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Leib, SL
影响因子：
3.3
作者：
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Bagetta, G.
影响因子：
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影响因子：
16.2
作者：
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通讯作者：
KOSHLAND, DE