Proteomic analysis of bronchoalveolar lavage fluid in rat exposed to TiO2 nanostructured aerosol by inhalation
吸入TiO2納米結(jié)構(gòu)氣溶膠大鼠支氣管肺泡灌洗液的蛋白質(zhì)組學(xué)分析
Laëtitia Chézeaua,b, Lori A. Kohlstaedtc, Alain Le Faoub, Frédéric Cosniera, Bertrand Rihnb,d, Laurent Gatéa,?
a Institut National de Recherche et de Sécurité, Rue du Morvan, CS 60027,Vandoeuvre, Cedex, France
b EA 3452 CITHEFOR, Université de Lorraine, BP 80403,Nancy Cedex, France
c California Institute for Quantitative Biosciences, University of California, Berkeley, CA 94720, United States of America
d Institut Jean-Lamour, UMR 7198 CNRS, Université de Lorraine,Nancy Cedex, France
A B S T R A C T
The pulmonary toxicological properties of inhaled titanium dioxide were studied using bronchoalveolar lavage fluid (BALF) cytology and proteomics analyses. Fischer 344 rats were exposed to 10 mg/m3 of TiO2 nanostructured aerosol by nose-only inhalation for 6 h/day, 5 days/week for 4 weeks. Lung samples were collected up to 180 post-exposure days. As previously described, cytological analyses of BALF showed a strong inflammatory response up to 3 post-exposure days, which persisted however, at a lower intensity up to 180 days. In addition, using Multidimensional Protein Identification Technology (MudPIT), we identified a total of 107, 50 and 45 proteins (UniprotKB identifiers) differentially expressed in exposed rats immediay, 3 and 180 days after the end of exposure respectively. Increased levels of inflammatory proteins, members of proteasome, various histones, proteins involved in cytoskeleton organization, were noticed up to 3 days (short-term response). Some of these proteins were linked with Neutrophil Extracellular Trap formation (NETosis). Long-term response was also characterized by a persistent altered expression of proteins up to 180 days. Altogether, these results suggest that exposure to low toxicity low solubility nanomaterials such as TiO2 may induce long-term changes in the pulmonary protein expression pattern of which the physio-pathological consequences are unknown. Significance: This paper describes in rats, at the pulmonary level, the effects of inhaled nanostructured aerosol of TiO2 on the secreted proteins found in the broncho-alveolar space by comparing the proteomic profile in broncho-alveolar lavage fluid supernatants of control and exposed animals. This work brings new insights about the early events occurring following the end of exposure and suggests the formation of Neutrophil Extracellular Traps (NETosis) that could be interpret as a potential early mechanism of defense against TiO2 nanoparticles. This work also describes the long term effects (180 post-exposure days) of such an exposure and the change in secreted protein expression in the absence of significant histopathological modifications.
用支氣管肺泡灌洗液(BALF)細胞學(xué)和蛋白質(zhì)組學(xué)分析研究吸入*的肺毒理特性。Fischer 344大鼠僅經(jīng)鼻吸入10mg/m3 TiO2納米結(jié)構(gòu)氣霧劑6小時/天，5天/周，連續(xù)4周。在暴露后180天內(nèi)采集肺樣本。如前所述，BALF的細胞學(xué)分析顯示，暴露后3天內(nèi)有強烈的炎癥反應(yīng)，但持續(xù)時間較低，達180天。此外，利用多維蛋白質(zhì)鑒定技術(shù)(MudPIT)，我們分別鑒定了暴露后即刻、3天和180天暴露大鼠體內(nèi)差異表達的107、50和45種蛋白質(zhì)(UniprotKB標(biāo)識符)。炎性蛋白、蛋白酶體成員、各種組蛋白、參與細胞骨架組織的蛋白水平增加，長3天(短期反應(yīng))。其中一些蛋白與中性粒細胞胞外陷阱形成(NETosis)有關(guān)。長期反應(yīng)還表現(xiàn)為持續(xù)180天的蛋白質(zhì)表達改變。總之，這些結(jié)果表明，暴露于低毒性的低溶解度納米材料如TiO2可誘導(dǎo)肺組織蛋白表達模式的長期變化，其中生理病理后果是未知的。意義：通過比較對照組和暴露組大鼠支氣管肺泡灌洗液上清液的蛋白質(zhì)組學(xué)特征，本文在大鼠肺水平上研究了吸入*納米結(jié)構(gòu)氣霧劑對支氣管肺泡腔分泌蛋白的影響。這項工作為暴露結(jié)束后發(fā)生的早期事件帶來了新的見解，并提出了中性粒細胞胞外陷阱(NETosis)的形成，可以解釋為對TiO2納米顆粒防御的潛在早期機制。這項工作還描述了這種暴露的長期影響(暴露后180天)以及在沒有明顯組織病理學(xué)改變的情況下分泌蛋白表達的變化。
Keywords: Titanium dioxide、Lung inflammation、Protein expression profile、Short-term response、Long-term response、NETosis
關(guān)鍵詞：*、肺部炎癥、蛋白表達譜、短期反應(yīng)、長期反應(yīng)、網(wǎng)狀內(nèi)皮細胞增生
