α1-antitrypsin modulates microglial-mediated neuroinflammation and protects microglial cells from amyloid-β-induced toxicity.
α1-antitrypsin modulates microglial-mediated neuroinflammation and protects microglial cells from amyloid-β-induced toxicity.
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DOI：
10.1186/s12974-014-0165-8
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发表时间：
2014-09-23
影响因子：
9.3
通讯作者：
Bach JP
中科院分区：
文献类型：
--
作者：
Gold M;Dolga AM;Koepke J;Mengel D;Culmsee C;Dodel R;Koczulla AR;Bach JP
阿尔茨海默病的一个标志是小胶质细胞活化。这种神经退行性疾病的治疗方法包括调节小胶质细胞。α1-抗胰蛋白酶（A1 AT）对巨噬细胞和肺上皮细胞具有抗炎作用，并抑制中性粒细胞中的钙蛋白酶活性。A1 AT对小胶质细胞介导的神经炎症的影响尚不清楚。本研究旨在探讨A1 AT对β淀粉样蛋白（Aβ）和脂多糖（LPS）处理的小胶质细胞的细胞因子产生、应激途径、细胞活力、吞噬能力的影响及其机制。从胚胎第13.5天的Swiss韦伯斯特小鼠胚胎中分离原代小胶质细胞。用ELISA分析处理的原代小胶质细胞上清液中的细胞因子，并用Griess试剂检测累积的亚硝酸盐。使用蛋白质印迹法在细胞裂解物中研究细胞内应激途径。在负载Ca 2+敏感（荧光）染料Fluo-4的BV-2小胶质细胞中检测到细胞内钙水平。通过使用钙蛋白酶活性测定来评估原代小胶质细胞中的钙蛋白酶活性。MTT法检测Aβ处理后小胶质细胞的存活率。用蛋白质印迹分析评估Aβ的吞噬作用。联合给药后，A1 AT可减少LPS或Aβ诱导的促炎介质。有趣的是，我们检测到钙蛋白酶活性和细胞内钙浓度的降低，这可能介导A1 AT的抗炎作用。经典活化途径的抑制，如促分裂原活化蛋白激酶的磷酸化或蛋白激酶A的活化被排除为A1 AT介导效应的机制。此外，A1 AT增加了Aβ处理的小胶质细胞的活力，降低了Aβ的吞噬作用。我们提供了证据的作用机制A1 AT对小胶质细胞介导的神经炎症在体外。我们的体外数据表明，A1 AT治疗调节炎症条件下的小胶质细胞，这种调节是由于钙蛋白酶活性和细胞内钙水平的抑制。这里观察到的影响的潜在机制是有前途的未来的治疗策略，因此，应进一步追求在阿尔茨海默病的转基因小鼠模型。
One hallmark of Alzheimer disease is microglial activation. Therapeutic approaches for this neurodegenerative disease include the modulation of microglial cells. α1-antitrypsin (A1AT) has been shown to exert anti-inflammatory effects on macrophages and lung epithelial cells and an inhibition of calpain activity in neutrophil granulocytes. Nothing is known about the effect of A1AT on microglial-mediated neuroinflammation. Our aim was to investigate the effect of A1AT on amyloid-β (Aβ)- and LPS-treated microglial cells in vitro with respect to cytokine production, stress pathways, cell viability, phagocytotic abilities and the underlying mechanisms. Primary microglial cells were isolated from Swiss Webster mouse embryos on embryonic day 13.5. Cytokines in the supernatants of treated primary microglial cells were analyzed with ELISAs, and accumulated nitrite was detected with Griess reagents. Intracellular stress pathways were investigated in cell lysates using western blotting. Intracellular calcium levels were detected in BV-2 microglial cells loaded with the Ca2+-sensitive (fluorescent) dye Fluo-4. Calpain activity in primary microglial cells was assessed by using a calpain activity assay. Cell viability of Aβ-treated microglial cells was analyzed using MTT assay. Phagocytosis of Aβ was evaluated with western blot analysis. Upon co-administration, A1AT reduced pro-inflammatory mediators induced by LPS or Aβ. Interestingly, we detected a reduction in calpain activity and in the concentration of intracellular calcium that might mediate the anti-inflammatory effects of A1AT. Inhibition of the classic activation pathways, such as phosphorylation of mitogen-activated protein kinases or activation of protein kinase A were excluded as a mechanism of A1AT-mediated effects. In addition, A1AT increased the viability of Aβ-treated microglial cells and reduced Aβ phagocytosis. We provide evidence on the mechanism of action of A1AT on microglial-mediated neuroinflammation in vitro. Our in vitro data indicate that A1AT treatment modulates microglial cells in inflammatory conditions and that this modulation is due to an inhibition of calpain activity and intracellular calcium levels. The underlying mechanisms of the effects observed here are promising for future therapeutic strategies and should thus be further pursued in transgenic mouse models of Alzheimer disease.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
2.9
作者：
Ii, M;Sunamoto, M;Ichimori, Y
通讯作者：
Ichimori, Y
影响因子：
4.4
作者：
Fettucciari, Katia;Fetriconi, Ilaria;Marconi, Pierfrancesco
通讯作者：
Marconi, Pierfrancesco
影响因子：
4.8
作者：
Janciauskiene, Sabina M.;Nita, Izabela M.;Stevens, Tim
通讯作者：
Stevens, Tim
DOI：
10.1073/pnas.90.7.2628
发表时间：
1993-04-01
影响因子：
11.1
作者：
SAITO, KI;ELCE, JS;NIXON, RA
通讯作者：
NIXON, RA