Emodin attenuates calcium overload and endoplasmic reticulum stress in AR42J rat pancreatic acinar cells

Wu,Li; Cai,Baochang; Liu,Xiao; Cai,Hao

doi:10.3892/mmr.2013.1773

Emodin attenuates calcium overload and endoplasmic reticulum stress in AR42J rat pancreatic acinar cells

Authors:
- Li Wu
- Baochang Cai
- Xiao Liu
- Hao Cai
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Affiliations: National First-Class Key Discipline for Science of Chinese Materia Medica, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China
Published online on: November 4, 2013 https://doi.org/10.3892/mmr.2013.1773
Pages: 267-272

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Abstract

The aim of the present study was to investigate the protective effects of emodin against calcium overload and endoplasmic reticulum (ER) stress in an acute pancreatitis model in vitro. AR42J rat pancreatic acinar cells treated with cerulein (10‑7 M) and lipoplysaccharide (LPS; 10 mg/l) were used to mimic acute pancreatitis in vitro. The amylase activity in cellular lysates and culture media was detected by spectrophotometry. The level of cytosolic calcium was measured by laser confocal microscopy. Cell apoptosis and necrosis were examined by flow cytometry. Reverse transcription‑polymerase chain reaction was used to determine the mRNA expression of ER chaperone immunoglobulin‑binding protein (Bip) and downstream molecules, including protein kinase‑like ER kinase (PERK), activation transcription factor 6 (ATF6) and inositol‑requiring protein 1 (IRE1). The results showed that emodin significantly reduced the expression and release of amylase, attenuated calcium overload and decreased the mRNA expression of Bip, PERK, ATF6 and IRE1 which was significantly elevated in AR42J cells treated with cerulein and LPS. Emodin also reduced cell apoptosis and necrosis. Therefore, the results of the present study indicate that emodin protects against AR42J cell injury caused by cerulein and LPS. These effects may be associated with reduced calcium overload and inhibited ER stress responses.

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1	Talukdar R and Swaroop Vege S: Early management of severe acute pancreatitis. Curr Gastroenterol Rep. 13:123–130. 2011. View Article : Google Scholar : PubMed/NCBI
2	Kylänpää ML, Repo H and Puolakkainen PA: Inflammation and immunosuppression in severe acute pancreatitis. World J Gastroenterol. 16:2867–2872. 2010.
3	Gorelick FS and Thrower E: The acinar cell and early pancreatitis responses. Clin Gastroenterol Hepatol. 7(11 Suppl): S10–S14. 2009. View Article : Google Scholar
4	Kim JY, Kim KH, Lee JA, Namkung W, Sun AQ, Ananthanarayanan M, Suchy FJ, Shin DM, Muallem S and Lee MG: Transporter-mediated bile acid uptake causes Ca²⁺-dependent cell death in rat pancreatic acinar cells. Gastroenterology. 122:1941–1953. 2002. View Article : Google Scholar
5	Criddle DN, Raraty MG, Neoptolemos JP, Tepikin AV, Petersen OH and Sutton R: Ethanol toxicity in pancreatic acinar cells: mediation by nonoxidative fatty acid metabolites. Proc Natl Acad Sci USA. 101:10738–10743. 2004. View Article : Google Scholar : PubMed/NCBI
6	Criddle DN, Murphy J, Fistetto G, Barrow S, Tepikin AV, Neoptolemos JP, Sutton R and Petersen OH: Fatty acid ethyl esters cause pancreatic calcium toxicity via inositol trisphosphate receptors and loss of ATP synthesis. Gastroenterology. 130:781–793. 2006. View Article : Google Scholar : PubMed/NCBI
7	Kubisch CH and Logsdon CD: Endoplasmic reticulum stress and the pancreatic acinar cell. Expert Rev Gastroenterol Hepatol. 2:249–260. 2008. View Article : Google Scholar : PubMed/NCBI
8	Kubisch CH, Sans MD, Arumugam T, Ernst SA, Williams JA and Logsdon CD: Early activation of endoplasmic reticulum stress is associated with arginine-induced acute pancreatitis. Am J Physiol Gastrointest Liver Physiol. 291:G238–G245. 2006. View Article : Google Scholar : PubMed/NCBI
9	Kubisch CH and Logsdon CD: Secretagogues differentially activate endoplasmic reticulum stress responses in pancreatic acinar cells. Am J Physiol Gastrointest Liver Physiol. 292:G1804–G1812. 2007. View Article : Google Scholar : PubMed/NCBI
10	Seyhun E, Malo A, Schäfer C, Moskaluk CA, Hoffmann RT, Göke B and Kubisch CH: Tauroursodeoxycholic acid reduces endoplasmic reticulum stress, acinar cell damage, and systemic inflammation in acute pancreatitis. Am J Physiol Gastrointest Liver Physiol. 301:G773–G782. 2011. View Article : Google Scholar : PubMed/NCBI
11	Szmola R and Sahin-Tóth M: Pancreatitis-associated chymotrypsinogen C (CTRC) mutant elicits endoplasmic reticulum stress in pancreatic acinar cells. Gut. 59:365–372. 2010. View Article : Google Scholar
12	Malo A, Krüger B, Seyhun E, Schäfer C, Hoffmann RT, Göke B and Kubisch CH: Tauroursodeoxycholic acid reduces endoplasmic reticulum stress, trypsin activation, and acinar cell apoptosis while increasing secretion in rat pancreatic acini. Am J Physiol Gastrointest Liver Physiol. 299:G877–G886. 2010. View Article : Google Scholar
13	Ye R, Mareninova OA, Barron E, Wang M, Hinton DR, Pandol SJ and Lee AS: Grp78 heterozygosity regulates chaperone balance in exocrine pancreas with differential response to cerulein-induced acute pancreatitis. Am J Pathol. 177:2827–2836. 2010. View Article : Google Scholar : PubMed/NCBI
14	Wang G, Sun B, Gao Y, Meng QH and Jiang HC: The effect of emodin-assisted early enteral nutrition on severe acute pancreatitis and secondary hepatic injury. Mediators Inflamm. 2007:296382007. View Article : Google Scholar : PubMed/NCBI
15	Li Z, Xia X, Zhang S, Zhang A, Bo W and Zhou R: Up-regulation of Toll-like receptor 4 was suppressed by emodin and baicalin in the setting of acute pancreatitis. Biomed Pharmacother. 63:120–128. 2009. View Article : Google Scholar : PubMed/NCBI
16	Xia XM, Li BK, Xing SM and Ruan HL: Emodin promoted pancreatic claudin-5 and occluding expression in experimental acute pancreatitis rats. World J Gastroenterol. 18:2132–2139. 2012. View Article : Google Scholar : PubMed/NCBI
17	Yu JH, Lim JW, Kim KH, Morio T and Kim H: NADPH oxidase and apoptosis in cerulein-stimulated pancreatic acinar AR42J cells. Free Radic Biol Med. 39:590–602. 2005. View Article : Google Scholar
18	Wu L, Cai B, Zheng S, Liu X, Cai H and Li H: Effect of emodin on endoplasmic reticulum stress in rats with severe acute pancreatitis. Inflammation. 36:1020–1029. 2013. View Article : Google Scholar : PubMed/NCBI
19	Eum WS, Li MZ, Sin GS, Choi SY, Park JB, Lee JY and Kwon HY: Dexamethasone-induced differentiation of pancreatic AR42J cell involves p21(waf1/cip1) and MAP kinase pathway. Exp Mol Med. 35:379–384. 2003. View Article : Google Scholar : PubMed/NCBI
20	Low JT, Shukla A and Thorn P: Pancreatic acinar cell: new insights into the control of secretion. Int J Biochem Cell Biol. 42:1586–1589. 2010. View Article : Google Scholar : PubMed/NCBI
21	Frick TW: The role of calcium in acute pancreatitis. Surgery. 152(Suppl 1): S157–S163. 2012. View Article : Google Scholar : PubMed/NCBI
22	Criddle DN, Gerasimenko JV, Baumgartner HK, Jaffar M, Voronina S, Sutton R, Petersen OH and Gerasimenko OV: Calcium signalling and pancreatic cell death: apoptosis or necrosis? Cell Death Differ. 14:1285–1294. 2007. View Article : Google Scholar : PubMed/NCBI
23	Criddle DN, Sutton R and Petersen OH: Role of Ca²⁺ in pancreatic cell death induced by alcohol metabolites. J Gastroenterol Hepatol. 21(Suppl 3): S14–S17. 2006.
24	Chen DC and Wang L: Mechanisms of therapeutic effects of rhubarb on gut origin sepsis. Chin J Traumatol. 12:365–369. 2009.PubMed/NCBI
25	Kuo YC, Meng HC and Tsai WJ: Regulation of cell proliferation, inflammatory cytokine production and calcium mobilization in primary human T lymphocytes by emodin from Polygonum hypoleucum Ohwi. Inflamm Res. 50:73–82. 2001. View Article : Google Scholar : PubMed/NCBI
26	Sui F, Huo HR, Zhang CB, Yang N, Guo JY, Du XL, Zhao BS, Liu HB, Li LF, Guo SY and Jiang TL: Emodin down-regulates expression of TRPV1 mRNA and its function in DRG neurons in vitro. Am J Chin Med. 38:789–800. 2010. View Article : Google Scholar : PubMed/NCBI
27	Liu Y, Zhou ZG, Chen KL, Zhou B, Yang L, Yan H and Li Y: The ER chaperone GRP78 is associated with the severity of cerulein-induced pancreatic inflammation via regulating apoptosis of pancreatic acinar cells. Hepatogastroenterology. 59:1670–1676. 2012.PubMed/NCBI
28	Petersen OH and Sutton R: Ca²⁺ signalling and pancreatitis: effects of alcohol, bile and coffee. Trends Pharmacol Sci. 27:113–120. 2006.
29	Kubisch CH and Logsdon CD: Secretagogues differentially activate endoplasmic reticulum stress responses in pancreatic acinar cells. Am J Physiol Gastrointest Liver Physiol. 292:G1804–G1812. 2007. View Article : Google Scholar
30	Pavitt GD and Ron D: New insights into translational regulation in the endoplasmic reticulum unfolded protein response. Cold Spring Harb Perspect Biol. 4:a0122782012. View Article : Google Scholar : PubMed/NCBI
31	Sendler M, Dummer A, Weiss FU, Krüger B, Wartmann T, Scharffetter-Kochanek K, van Rooijen N, Malla SR, Aghdassi A, Halangk W, Lerch MM and Mayerle J: Tumour necrosis factor α secretion induces protease activation and acinar cell necrosis in acute experimental pancreatitis in mice. Gut. 62:430–439. 2013.
32	Maguire JA, Mulugeta S and Beers MF: Endoplasmic reticulum stress induced by surfactant protein C Brichos mutants promotes proinflammatory signaling by epithelial cells. Am J Respir Cell Mol Biol. 44:404–414. 2011. View Article : Google Scholar
33	Kim SM, Chung MJ, Ha TJ, Choi HN, Jang SJ, Kim SO, Chun MH, Do SI, Choo YK and Park YI: Neuroprotective effects of black soybean anthocyanins via inactivation of ASK1-JNK/p38 pathways and mobilization of cellular sialic acids. Life Sciences. 90:874–882. 2012. View Article : Google Scholar : PubMed/NCBI