流行病學研究顯示空氣污染與代謝症候群發展有關，且在動物實驗中發現細懸浮微粒 (粒徑≤ 2.5μm,PM2.5)可能產生肝臟毒性，並促進發炎、脂肪變性和降低肝醣儲存等類酒精性脂肪肝之症狀。有研究指出半乳醣凝集素-3會與胰島素受體結合並導致胰島素阻抗的發展。本研究探討亞慢性超細微碳黑呼吸暴露對於大鼠肝臟的影響，是否會改變肝細胞中肝醣的儲存功能以及是否有脂肪堆積於肝臟細胞內，觀察暴露超細微碳黑後肝臟胰島素敏感性的改變，並探討半乳醣凝集素-3於肝臟的表現。
大鼠暴露於數目濃度為六十萬顆/立方公分的超細微碳黑，一天6小時，一週5天，總共10週或13週。暴露結束後一天將大鼠犧牲並使用蘇木精-伊紅素染色觀察形態組織變化，以免疫組織化學染色法觀察巨噬細胞生物標誌物CD68以及半乳醣凝集素-3的表現量。以西式墨點法測定肝臟中半乳醣凝集素-3、p-Akt及Akt之表現量。
結果發現，暴露超細微碳黑13週，在肝門靜脈周圍有發炎細胞聚集和脂肪在細胞中過度堆積，但並未觀察到有氣球狀變性，利用免疫組織化學染色法發現暴露超細微碳黑的組別中CD68及半乳醣凝集素-3的表現量高於控制組。在馬森三色染色法中觀察到在肝門靜脈及肝門管區膠原表現有增加之情形，尚未有纖維化之表現，而透過PAS染色發現暴露超細微碳黑肝醣含量相對較少。西式墨點實驗結果顯示暴露超細微碳黑之組別相對過濾空氣組與管錐控制組的半乳醣凝集素-3蛋白表現量有顯著增加之情形，而p-Akt/Akt之蛋白表現有顯著減少。
暴露超細微碳黑微粒後會造成肝臟組織病變，且有發炎以及脂肪變性，且有肝醣儲存下降之情形。在西方墨點法檢測發現半乳醣凝集素-3顯著上升、p-Akt/Akt顯著下降。因此，亞慢性超細微碳黑呼吸暴露對於大鼠肝臟會導致組織發炎、肝細胞肝醣儲存功能下降及胰島素敏感性的改變。

SUMMARY
The purpose of this study was to evaluate pathological changes in the liver of animals exposed to ultrafine carbon black (ufCB). Male Wistar rats were exposed to ultrafine carbon black for 6hrs/day, 5days/week, for 10weeks or 13weeks using computerized nose-only inhalation system. Histopathological evaluation showed Kupffer cell infiltration and hepatic steatosis but no fibrosis observed. In addition, rat displayed impaired hepatic glycogen storage. Hepatic Gal-3 was increased significantly and p-Akt/Akt ratio was significantly decreased in ufCB group compared with those in FA group and Tube control group. Our results found that subchronic inhalation exposed to ufCB may induce hepatic inflammation, impair hepatic glycogen storage, and the increased Gal-3 may play an important role in development of insulin resistance.

Key words：ultrafine carbon black, liver, Gal-3, pathological change

Alberti KGM, Zimmet P, Shaw J. 2005. The metabolic syndrome—a new worldwide definition. The Lancet 366:1059-1062.
Anderson JO, Thundiyil JG, Stolbach A. 2012. Clearing the air: A review of the effects of particulate matter air pollution on human health. Journal of Medical Toxicology 8:166-175.
Angluo P. 2002. Nonalcoholic fatty liver disease. N Engl J Med 346:1221-1231.
Arciello M, Gori M, Maggio R, Barbaro B, Tarocchi M, Galli A, et al. 2013. Environmental pollution: A tangible risk for nafld pathogenesis. International Journal of Molecular Sciences 14:22052-22066.
Argo CK, Northup PG, Al-Osaimi AM, Caldwell SH. 2009. Systematic review of risk factors for fibrosis progression in non-alcoholic steatohepatitis. Journal of Hepatology 51:371-379.
Arul D, Subramanian P. 2014. Daytime food restriction influences the circadian rhythms of circulatory hepatic marker enzymes activity in rats. Biological Rhythm Research 45:643-652.
Bedogni G, Bellentani S, Miglioli L, Masutti F, Passalacqua M, Castiglione A, et al. 2006. The fatty liver index: A simple and accurate predictor of hepatic steatosis in the general population. BMC Gastroenterology 6:33.
Beelen R, Hoek G, van den Brandt PA, Goldbohm RA, Fischer P, Schouten LJ, et al. 2008. Long-term exposure to traffic-related air pollution and lung cancer risk. Epidemiology 19:702-710.
Bellentani S, Scaglioni F, Marino M, Bedogni G. 2010. Epidemiology of non-alcoholic fatty liver disease. Digestive Diseases 28:155-161.
Brauer M, Hoek G, Van Vliet P, Meliefste K, Fischer PH, Wijga A, et al. 2002. Air pollution from traffic and the development of respiratory infections and asthmatic and allergic symptoms in children. American Journal of Respiratory and Critical Care Medicine 166:1092-1098.
Brook RD, Jerrett M, Brook JR, Bard RL, Finkelstein MM. 2008. The relationship between diabetes mellitus and traffic-related air pollution. Journal of Occupational and Environmental Medicine 50:32-38.
Brunt EM, Janney CG, Di Bisceglie AM, Neuschwander-Tetri BA, Bacon BR. 1999. Nonalcoholic steatohepatitis: A proposal for grading and staging the histological lesions. The American Journal of Gastroenterology 94:2467-2474.
Brunt EM. 2010. Pathology of nonalcoholic fatty liver disease. Nature Reviews Gastroenterology & Hepatology 7:195-203.
Caldwell S, Ikura Y, Dias D, Isomoto K, Yabu A, Moskaluk C, et al. 2010. Hepatocellular ballooning in nash. Journal of Hepatology 53:719-723.
Chen J-C, Schwartz J. 2008. Metabolic syndrome and inflammatory responses to long-term particulate air pollutants. Environmental Health Perspectives 116:612-617.
Chu C, Zhou L, Xie H, Pei Z, Zhang M, Wu M, et al. 2019. Pulmonary toxicities from a 90-day chronic inhalation study with carbon black nanoparticles in rats related to the systemical immune effects. International Journal of Nanomedicine 14:2995.
Day CP, James OF. 1998. Steatohepatitis: A tale of two “hits”? :Elsevier.
Denk H, Stumptner C, Zatloukal K. 2000. Mallory bodies revisited. Journal of Hepatology 32:689-702.
Díaz-Muñoz M, Vázquez-Martínez O, Báez-Ruiz A, Martínez-Cabrera G, Soto-Abraham MV, Ávila-Casado MC, et al. 2010. Daytime food restriction alters liver glycogen, triacylglycerols, and cell size. A histochemical, morphometric, and ultrastructural study. Comparative Hepatology 9:1-10.
Eze IC, Schaffner E, Foraster M, Imboden M, von Eckardstein A, Gerbase MW, et al. 2015. Long-term exposure to ambient air pollution and metabolic syndrome in adults. PLoS One 10.
Farrell GC, Chitturi S, Lau GK, Sollano JD, NAFLD APWPo. 2007. Guidelines for the assessment and management of non‐alcoholic fatty liver disease in the asia–pacific region: Executive summary. Journal of Gastroenterology and Hepatology 22:775-777.
Filep Á, Fodor GH, Kun-Szabó F, Tiszlavicz L, Rázga Z, Bozsó G, et al. 2016. Exposure to urban pm1 in rats: Development of bronchial inflammation and airway hyperresponsiveness. Respiratory Research 17:26.
Fritsch K, Mernberger M, Nist A, Stiewe T, Brehm A, Jacob R. 2016. Galectin-3 interacts with components of the nuclear ribonucleoprotein complex. BMC Cancer 16:1-10.
Grunig G, Marsh LM, Esmaeil N, Jackson K, Gordon T, Reibman J, et al. 2014. Perspective: Ambient air pollution: Inflammatory response and effects on the lung's vasculature. Pulmonary Circulation 4:25-35.
Hoek G, Brunekreef B, Goldbohm S, Fischer P, van den Brandt PA. 2002. Association between mortality and indicators of traffic-related air pollution in the netherlands: A cohort study. The Lancet 360:1203-1209.
Hsu C-S, Kao J-H. 2012. Non-alcoholic fatty liver disease: An emerging liver disease in taiwan. Journal of the Formosan Medical Association 111:527-535.
Iacobini C, Menini S, Ricci C, Fantauzzi CB, Scipioni A, Salvi L, et al. 2011. Galectin-3 ablation protects mice from diet-induced nash: A major scavenging role for galectin-3 in liver. Journal of Hepatology 54:975-983.
Invernizzi G, Ruprecht A, Mazza R, De Marco C, Močnik G, Sioutas C, et al. 2011. Measurement of black carbon concentration as an indicator of air quality benefits of traffic restriction policies within the ecopass zone in milan, italy. Atmospheric Environment 45:3522-3527.
Ipsen DH, Lykkesfeldt J, Tveden-Nyborg P. 2018. Molecular mechanisms of hepatic lipid accumulation in non-alcoholic fatty liver disease. Cellular and Molecular Life Sciences 75:3313-3327.
Jackson P, Vogel U, Wallin H, Hougaard KS. 2011. Maternal exposure to particulate air pollution and engineered nanoparticles: Reproductive and Developmental Effects. Air Pollution-New Developments Rijeka University Campus Step Ri:59-84.
Kang HG, Kim D-H, Kim S-J, Cho Y, Jung J, Jang W, et al. 2016. Galectin-3 supports stemness in ovarian cancer stem cells by activation of the notch1 intracellular domain. Oncotarget 7:68229.
Kim JW, Park S, Lim CW, Lee K, Kim B. 2014. The role of air pollutants in initiating liver disease. Toxicological Research 30:65-70.
Kleiner DE, Brunt EM, Van Natta M, Behling C, Contos MJ, Cummings OW, et al. 2005. Design and validation of a histological scoring system for nonalcoholic fatty liver disease. Hepatology 41:1313-1321.
Lackner C. 2011. Hepatocellular ballooning in nonalcoholic steatohepatitis: The pathologist’s perspective. Expert Review of Gastroenterology & Hepatology 5:223-231.
Li P, Lu M, Nguyen MA, Bae EJ, Chapman J, Feng D, et al. 2010. Functional heterogeneity of cd11c-positive adipose tissue macrophages in diet-induced obese mice. Journal of Biological Chemistry 285:15333-15345.
Li P, Liu S, Lu M, Bandyopadhyay G, Oh D, Imamura T, et al. 2016. Hematopoietic-derived galectin-3 causes cellular and systemic insulin resistance. Cell 167:973-984. e912.
Li W, Dorans KS, Wilker EH, Rice MB, Long MT, Schwartz J, et al. 2017. Residential proximity to major roadways, fine particulate matter, and hepatic steatosis: The framingham heart study. American Journal of Epidemiology 186:857-865.
Liu C, Xu X, Bai Y, Wang T-Y, Rao X, Wang A, et al. 2014. Air pollution–mediated susceptibility to inflammation and insulin resistance: Influence of ccr2 pathways in mice. Environmental Health Perspectives 122:17-26.
Marchesini G, Brizi M, Morselli-Labate AM, Bianchi G, Bugianesi E, McCullough AJ, et al. 1999. Association of nonalcoholic fatty liver disease with insulin resistance. The American Journal of Medicine 107:450-455.
Matteoni CA, Younossi ZM, Gramlich T, Boparai N, Liu YC, McCullough AJ. 1999. Nonalcoholic fatty liver disease: A spectrum of clinical and pathological severity. Gastroenterology 116:1413-1419.
Meshkani R, Adeli K. 2009. Hepatic insulin resistance, metabolic syndrome and cardiovascular disease. Clinical Biochemistry 42:1331-1346.
National GCU. 2016. Non-alcoholic fatty liver disease: Assessment and management.
Neuschwander-Tetri BA. 2017. Non-alcoholic fatty liver disease. BMC Medicine 15:45.
Niwa Y, Hiura Y, Sawamura H, Iwai N. 2008. Inhalation exposure to carbon black induces inflammatory response in rats. Circulation Journal 72:144-149.
Patsouris D, Li P-P, Thapar D, Chapman J, Olefsky JM, Neels JG. 2008. Ablation of cd11c-positive cells normalizes insulin sensitivity in obese insulin resistant animals. Cell Metabolism 8:301-309.
Petersen KF, Dufour S, Befroy D, Lehrke M, Hendler RE, Shulman GI. 2005. Reversal of nonalcoholic hepatic steatosis, hepatic insulin resistance, and hyperglycemia by moderate weight reduction in patients with type 2 diabetes. Diabetes 54:603-608.
Pope 3rd C. 2000. Epidemiology of fine particulate air pollution and human health: Biologic mechanisms and who's at risk? Environmental Health Perspectives 108:713.
Puri P, Sanyal AJ. 2012. Nonalcoholic fatty liver disease: Definitions, risk factors, and workup. Clinical Liver Disease 1:99.
Rachek LI. 2014. Free fatty acids and skeletal muscle insulin resistance. In: Progress in molecular biology and translational science, Vol. 121:Elsevier, 267-292.
Sato S, Bhaumik P, St-Pierre G, Pelletier I. 2014. Role of galectin-3 in the initial control of leishmania infection. Critical Reviews™ in Immunology 34.
Schomburg L. 2017. Plant-based diets and selenium intake and status. In: Vegetarian and plant-based diets in health and disease prevention:Elsevier, 729-746.
Skoien R, Richardson MM, Jonsson JR, Powell EE, Brunt EM, Neuschwander‐Tetri BA, et al. 2013. Heterogeneity of fibrosis patterns in non‐alcoholic fatty liver disease supports the presence of multiple fibrogenic pathways. Liver International 33:624-632.
Slezakova K, Morais S, do Carmo Pereira M. 2013. Atmospheric nanoparticles and their impacts on public health. In: Current topics in public health:IntechOpen.
Sun L, Liu C, Xu X, Ying Z, Maiseyeu A, Wang A, et al. 2013. Ambient fine particulate matter and ozone exposures induce inflammation in epicardial and perirenal adipose tissues in rats fed a high fructose diet. Particle and Fibre Toxicology 10:1-20.
Sun Q, Yue P, Deiuliis JA, Lumeng CN, Kampfrath T, Mikolaj MB, et al. 2009. Ambient air pollution exaggerates adipose inflammation and insulin resistance in a mouse model of diet-induced obesity. Circulation 119.
Sun Q, Zhang G, Chen R, Li R, Wang H, Jiang A, et al. 2018. Central ikk2 inhibition ameliorates air pollution-mediated hepatic glucose and lipid metabolism dysfunction in mice with type ii diabetes. Toxicological Sciences 164:240-249.
Takahashi Y, Fukusato T. 2014. Histopathology of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis. World Journal of Gastroenterology: WJG 20:15539.
Tan H-H, Fiel MI, Sun Q, Guo J, Gordon RE, Chen L-C, et al. 2009. Kupffer cell activation by ambient air particulate matter exposure may exacerbate non-alcoholic fatty liver disease. Journal of Immunotoxicology 6:266-275.
Targher G, Day CP, Bonora E. 2010. Risk of cardiovascular disease in patients with nonalcoholic fatty liver disease. New England Journal of Medicine 363:1341-1350.
Tomaru M, Takano H, Inoue K-I, Yanagisawa R, Osakabe N, Yasuda A, et al. 2007. Pulmonary exposure to diesel exhaust particles enhances fatty change of the liver in obese diabetic mice. International Journal of Molecular Medicine 19:17-22.
Turchinovich A, Baranova A, Drapkina O, Tonevitsky A. 2018. Cell-free circulating nucleic acids as early biomarkers for nafld and nafld-associated disorders. Frontiers in Physiology 9:1256.
Vernon G, Baranova A, Younossi Z. 2011. Systematic review: The epidemiology and natural history of non‐alcoholic fatty liver disease and non‐alcoholic steatohepatitis in adults. Alimentary Pharmacology & Therapeutics 34:274-285.
Vuppalanchi R, Chalasani N. 2009. Nonalcoholic fatty liver disease and nonalcoholic steatohepatitis: Selected practical issues in their evaluation and management. Hepatology 49:306-317.
WHO. 2013. Health effects of particulate matter. Policy implications for countries in eastern europe, caucasus and central asia. Copenhagen: Who regional office for europe; 2013.
Xu J, Zhang W, Lu Z, Zhang F, Ding W. 2017. Airborne pm2. 5-induced hepatic insulin resistance by nrf2/jnk-mediated signaling pathway. International Journal of Environmental Research and Public Health 14:787.
Xu X, Yavar Z, Verdin M, Ying Z, Mihai G, Kampfrath T, et al. 2010. Effect of early particulate air pollution exposure on obesity in mice: Role of p47phox. Arteriosclerosis, Thrombosis, and Vascular Biology 30:2518-2527.
Xu X, Liu C, Xu Z, Tzan K, Zhong M, Wang A, et al. 2011. Long-term exposure to ambient fine particulate pollution induces insulin resistance and mitochondrial alteration in adipose tissue. Toxicological Sciences 124:88-98.
Younossi ZM. 2019. Non-alcoholic fatty liver disease–a global public health perspective. Journal of Hepatology 70:531-544.
Zatloukal K, Stumptner C, Fuchsbichler A, Fickert P, Lackner C, Trauner M, et al. 2004. The keratin cytoskeleton in liver diseases. The Journal of Pathology: A Journal of the Pathological Society of Great Britain and Ireland 204:367-376.
Zheng Z, Xu X, Zhang X, Wang A, Zhang C, Hüttemann M, et al. 2013. Exposure to ambient particulate matter induces a nash-like phenotype and impairs hepatic glucose metabolism in an animal model. Journal of Hepatology 58:148-154.
Zheng Z, Zhang X, Wang J, Dandekar A, Kim H, Qiu Y, et al. 2015. Exposure to fine airborne particulate matters induces hepatic fibrosis in murine models. Journal of Hepatology 63:1397-1404.