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Abstract: Objectives: Severe acute pancreatitis (SAP) can lead to acute lung injury (ALI). The purpose of this paper is to investigate the protective effect of clodronate-containing liposomes on ALI in rats with SAP. Methods: The thin film method was used to prepare liposomes. Sprague-Dawley rats were randomly divided into three groups. After the SAP model was established by injecting 5% (w/v) sodium taurocholate (2 ml/kg body weight) into the subcapsular space of the pancreata, normal saline was administered to the control (C) group, phosphate buffer solution (PBS)-containing liposome to the P group, and clodronate-containing liposome to the T group through tail veins. Blood samples were obtained from the superior mesenteric vein at 2 and 6 h to measure the levels of amylase, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Morphological changes in the pancreata and lung were observed using hematoxylin and eosin (H&E) staining, while cell apoptosis was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). In addition, the macrophage marker cluster of differentiation 68 (CD68) in lung tissue was detected with immunohistochemistry. Results: Blood levels of amylase, IL-6, and TNF-α were significantly increased in the P group compared to those in the T group (P<0.05). In the T group, large numbers of TUNEL-positive cells were observed, but no or few in the C and P groups. Gross inspection and H&E staining of pancreata and lung showed dramatic tissue damage, including inflammation and necrosis in the P group. Less remarkable changes were noted in the T group, and the C group exhibited normal histology. The histological scores according to Kaiser’s criteria were consistent with H&E findings. The number of CD68-positive macrophages decreased in the T group. Conclusions: Clodronate-containing liposomes have a protective effect against ALI in rats with SAP. Blockade of macrophages may represent a novel therapeutic strategy in SAP.

[1]Ammori, B.J., 2003. Role of the gut in the course of severe acute pancreatitis. Pancreas, 26(2):122-129.

[2]Bem, R.A., Farnand, A.W., Wong, V., Koski, A., Rosenfeld, M.E., van Rooijen, N., Frevert, C.W., Martin, T.R., Matute-Bello, G., 2008. Depletion of resident alveolar macrophages does not prevent Fas-mediated lung injury in mice. Am. J. Physiol. Lung Cell Mol. Physiol., 295(2):L314-L325.

[3]Chen, X., Li, S.L., Wu, T., Liu, J.D., 2008. Proteasome inhibitor ameliorates severe acute pancreatitis and associated lung injury of rats. World J. Gastroenterol., 14(20):3249-3253.

[4]Cheng, S., He, S., Zhang, J., 2002. The role of alveolar macrophage activation in rats with lung injury associated with acute necrotizing pancreatitis. Zhonghua Wai Ke Za Zhi, 40(8):609-612 (in Chinese).

[5]Closa, D., Sabater, L., Fernandez-Cruz, L., Prats, N., Gelpi, E., Roselló-Catafau, J., 1999. Activation of alveolar macrophages in lung injury associated with experimental acute pancreatitis is mediated by the liver. Ann. Surg., 229(2):230-236.

[6]Danenberg, H.D., Fishbein, I., Gao, J., Mönkkönen, J., Reich, R., Gati, I., Moerman, E., Golomb, G., 2002. Macrophage depletion by clodronate-containing liposomes reduces neointimal formation after balloon injury in rats and rabbits. Circulation, 106(5):599-605.

[7]Folch-Puy, E., 2007. Importance of the liver in systemic complications associated with acute pancreatitis: the role of Kupffer cells. J. Pathol., 211(4):383-388.

[8]Frith, J.C., Monkkonen, J., Blackburn, G.M., Russell, R.G., Rogers, M.J., 1997. Clodronate and liposome-encapsulated clodronate are metabolized to a toxic ATP analog, adenosine 5′-(β,γ-dichloromethylene) triphosphate, by mammalian cells in vitro. J. Bone Miner. Res., 12(9):1358-1367.

[9]Gutierrez, P.T., Folch-Puy, E., Bulbena, O., Closa, D., 2008. Oxidised lipids present in ascitic fluid interfere with the regulation of the macrophages during acute pancreatitis, promoting an exacerbation of the inflammatory response. Gut, 57(5):642-648.

[10]Kaiser, A.M., Saluja, A.K., Sengupta, A., Saluja, M., Steer, M.L., 1995. Relationship between severity, necrosis, and apoptosis in five models of experimental acute pancreatitis. Am. J. Physiol. Cell Physiol., 269 (5 Pt. 1):C1295-C1304.

[11]Lankisch, P.G., Lerch, M.M., 2006. Pharmacological prevention and treatment of acute pancreatitis: where are we now? Dig. Dis., 24(1-2):148-159.

[12]Lee, J., Kwon, W., Jo, Y., Suh, G., Youn, Y., 2008. Protective effects of ethyl pyruvate treatment on paraquat-intoxicated rats. Hum. Exp. Toxicol., 27(1):49-54.

[13]Makhija, R., Kingsnorth, A.N., 2002. Cytokine storm in acute pancreatitis. J. Hepatobiliary Pancreat. Surg., 9(4):401-410.

[14]Matsuda, N., Nishihira, J., Takahashi, Y., Kemmotsu, O., Hattori, Y., 2006. Role of macrophage migration inhibitory factor in acute lung injury in mice with acute pancreatitis complicated by endotoxemia. Am. J. Respir. Cell Mol. Biol., 35(2):198-205.

[15]Pezzilli, R., Bellacosa, L., Felicani, C., 2009. Lung injury in acute pancreatitis. JOP, 10(5):481-484.

[16]Roelofs, A.J., Thompson, K., Gordon, S., Rogers, M.J., 2006. Molecular mechanisms of action of bisphosphonates: current status. Clin. Cancer Res., 12(20):6222s-6230s.

[17]Selander, K.S., Monkkonen, J., Karhukorpi, E.K., Harkonen, P., Hannuniemi, R., Vaananen, H.K., 1996. Characteristics of clodronate-induced apoptosis in osteoclasts and macrophages. Mol. Pharmacol., 50(5):1127-1138.

[18]Serin, M., Gulbas, H., Gurses, I., Erkal, H.S., Yucel, N., 2007. The histopathological evaluation of the effectiveness of melatonin as a protectant against acute lung injury induced by radiation therapy in a rat model. Int. J. Radiat. Biol., 83(3):187-193.

[19]Sha, H., Ma, Q., Jha, R.K., Wang, Z., 2009. Resveratrol ameliorates lung injury via inhibition of apoptosis in rats with severe acute pancreatitis. Exp. Lung Res., 35(4):344-358.

[20]Singh, V.K., Wu, B.U., Bollen, T.L., Repas, K., Maurer, R., Mortele, K.J., Banks, P.A., 2009. Early systemic inflammatory response syndrome is associated with severe acute pancreatitis. Clin. Gastroenterol. Hepatol., 7(11):1247-1251.

[21]Steer, M.L., 2001. Relationship between pancreatitis and lung diseases. Respir. Physiol., 128(1):13-16.

[22]Sugita, H., Yamaguchi, Y., Ikei, S., Yamada, S., Ogawa, M., 1997. Enhanced expression of cytokine-induced neutrophil chemoattractant (CINC) by bronchoalveolar macrophages in cerulein-induced pancreatitis rats. Dig. Dis. Sci., 42(1):154-160.

[23]Traeger, T., Kessler, W., Hilpert, A., Mikulcak, M., Entleutner, M., Koerner, P., Westerholt, A., Cziupka, K., van Rooijen, N., Heidecke, C.D., et al., 2009. Selective depletion of alveolar macrophages in polymicrobial sepsis increases lung injury, bacterial load and mortality but does not affect cytokine release. Respiration, 77(2):203-213.

[24]van Rooijen, N., Sanders, A., 1994. Liposome mediated depletion of macrophages: mechanism of action, preparation of liposomes and applications. J. Immunol. Methods, 174(1-2):83-93.

[25]van Rooijen, N., van Kesteren-Hendrikx, E., 2003. “In vivo” depletion of macrophages by liposome-mediated “suicide”. Methods Enzymol., 373:3-16.

[26]van Rooijen, N., Sanders, A., van den Berg, T.K., 1996. Apoptosis of macrophages induced by liposome-mediated intracellular delivery of clodronate and propamidine. J. Immunol. Methods, 193(1):93-99.

[27]Viterbo, D., Bluth, M.H., Mueller, C.M., Zenilman, M.E., 2008. Mutational characterization of pancreatitis-associated protein 2 domains involved in mediating cytokine secretion in macrophages and the NF-κB pathway. J. Immunol., 181(3):1959-1968.

[28]Weinbroum, A.A., 2009. Mannitol prevents acute lung injury after pancreas ischemia-reperfusion: a dose-response, ex vivo study. Lung, 187(4):215-224.

[29]Zhang, J.X., Dang, S.C., 2006. Ligustrazine alleviates acute lung injury in a rat model of acute necrotizing pancreatitis. Hepatobiliary Pancreat. Dis. Int., 5(4):605-609.

[30]Zhang, J.X., Dang, S.C., Qu, J.G., Wang, X.Q., 2006. Preventive effect of tetramethylpyrazine on intestinal mucosal injury in rats with acute necrotizing pancreatitis. World J. Gastroenterol., 12(39):6386-6390.

[31]Zhang, J.X., Dang, S.C., Zhang, Y., Sha, X., Zhang, L.R., Wei, C.S., Chen, M., Jiang, D.L., 2010. MRI shows clodronate-liposomes attenuating liver injury in rats with severe acute pancreatitis. Hepatobiliary Pancreat. Dis. Int., 9(2):192-200.

[32]Zhao, M.Q., Fernandez, L.G., Doctor, A., Sharma, A.K., Zarbock, A., Tribble, C.G., Kron, I.L., Laubach, V.E., 2006. Alveolar macrophage activation is a key initiation signal for acute lung ischemia-reperfusion injury. Am. J. Physiol. Lung Cell Mol. Physiol., 291(5):L1018-L1026.