Pre-treatment of recombinant mouse MFG-E8 downregulates LPS-induced TNF-α production in macrophages via STAT3-mediated SOCS3 activation.
Pre-treatment of recombinant mouse MFG-E8 downregulates LPS-induced TNF-α production in macrophages via STAT3-mediated SOCS3 activation.
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DOI：
10.1371/journal.pone.0027685
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Wang P
中科院分区：
文献类型：
--
作者：
Aziz M;Jacob A;Matsuda A;Wu R;Zhou M;Dong W;Yang WL;Wang P
乳脂肪球-表皮生长因子 8 (MFG-E8) 通过调节细胞信号传导来调节先天免疫功能，但对此了解较少。在此，我们的目的是通过用重组鼠 MFG-E8 (rmMFG-E8) 预处理，然后用 LPS 刺激从野生型 (WT) 或 MFG-E8−/− 小鼠分离的 RAW264.7 细胞和腹膜巨噬细胞，研究 MFG-E8 在巨噬细胞中的直接抗炎作用。经rmMFG-E8处理的RAW264.7细胞和小鼠腹腔巨噬细胞，LPS诱导的TNF-α mRNA水平分别显着下调25%和24%，蛋白质水平分别显着下调29%和23%（P<0.05）。相反，与用 LPS 处理的 WT 小鼠相比，从 MFG-E8−/− 小鼠中分离的腹膜巨噬细胞产生的 TNF-α 水平高出 28%。在体内，腹腔注射LPS（5 mg/kg BW）诱导内毒素血症，诱导后4 h，MFG-E8−/−小鼠血清TNF-α水平（837 pg/mL）显着高于WT小鼠（570 pg/mL，P<0.05）。为了阐明 MFG-E8 的直接抗炎作用，我们检查了 STAT3 及其靶基因 SOCS3。 rmMGF-E8 处理显着诱导巨噬细胞中的 pSTAT3 和 SOCS3。在腹膜细胞和脾组织中 rmMFG-E8 的体内处理中观察到类似的结果。 rmMFG-E8 预处理显着降低了 LPS 诱导的 NF-κB p65 含量。这些数据清楚地表明，rmMFG-E8 上调 SOCS3，进而与 NF-κB p65 相互作用，促进 TLR4 信号传导对 LPS 诱导的 TNF-α 产生的负调节。我们的研究结果强烈表明 MFG-E8 是一种直接抗炎分子，并且可以将其开发为减轻炎症和组织损伤的疗法。
Milk fat globule-epidermal growth factor factor 8 (MFG-E8) regulates innate immune function by modulating cellular signaling, which is less understood. Herein, we aimed to investigate the direct anti-inflammatory role of MFG-E8 in macrophages by pre-treatment with recombinant murine MFG-E8 (rmMFG-E8) followed by stimulation with LPS in RAW264.7 cells and in peritoneal macrophages, isolated from wild-type (WT) or MFG-E8−/− mice. RAW264.7 cells and mouse peritoneal macrophages treated with rmMFG-E8 significantly downregulated LPS-induced TNF-α mRNA by 25% and 24%, and protein levels by 29% and 23%, respectively (P<0.05). Conversely, peritoneal macrophages isolated from MFG-E8−/− mice produced 28% higher levels of TNF-α, as compared to WT mice when treated with LPS. In in vivo, endotoxemia induced by intraperitoneal injection of LPS (5 mg/kg BW), at 4 h after induction, serum level of TNF-α was significantly higher in MFG-E8−/− mice (837 pg/mL) than that of WT (570 pg/mL, P<0.05). To elucidate the direct anti-inflammatory effect of MFG-E8, we examined STAT3 and its target gene, SOCS3. Treatment with rmMGF-E8 significantly induced pSTAT3 and SOCS3 in macrophages. Similar results were observed in in vivo treatment of rmMFG-E8 in peritoneal cells and splenic tissues. Pre-treatment with rmMFG-E8 significantly reduced LPS-induced NF-κB p65 contents. These data clearly indicated that rmMFG-E8 upregulated SOCS3 which in turn interacted with NF-κB p65, facilitating negative regulation of TLR4 signaling for LPS-induced TNF-α production. Our findings strongly suggest that MFG-E8 is a direct anti-inflammatory molecule, and that it could be developed as a therapy in attenuating inflammation and tissue injury.
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DOI：
10.4049/jimmunol.182.1.581
发表时间：
2009-01-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Komura H;Miksa M;Wu R;Goyert SM;Wang P
通讯作者：
Wang P
影响因子：
11.2
作者：
Jinushi, Masahisa;Nakazaki, Yukoh;Dranoff, Glenn
通讯作者：
Dranoff, Glenn
影响因子：
64.5
作者：
Dinarello CA
通讯作者：
Dinarello CA
影响因子：
3.1
作者：
Lin, WJ;Yeh, WC
通讯作者：
Yeh, WC
影响因子：
4.1
作者：
Bhattacharya, Sujoy;Ray, Ramesh M.;Johnson, Leonard R.
通讯作者：
Johnson, Leonard R.