Elimination of intracellular Ca<sup>2+</sup> overload by BAPTA‑AM liposome nanoparticles: A promising treatment for acute pancreatitis

Fu,Zailin; Fu,Zailin; Wang,Dingsheng; Wang,Dingsheng; Zheng,Caiyun; Zheng,Caiyun; Xie,Minghua; Xie,Minghua; Chen,Yifang; Chen,Yifang; Zhou,Yi; Zhou,Yi; Huang,Yan; Huang,Yan; Song,Ying; Song,Ying; Hong,Weiyong; Hong,Weiyong

doi:10.3892/ijmm.2024.5358

Elimination of intracellular Ca²⁺ overload by BAPTA‑AM liposome nanoparticles: A promising treatment for acute pancreatitis

Authors:
- Zailin Fu
- Dingsheng Wang
- Caiyun Zheng
- Minghua Xie
- Yifang Chen
- Yi Zhou
- Yan Huang
- Ying Song
- Weiyong Hong
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Affiliations: Department of Pharmacy, Linping Branch, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310000, P.R. China, Department of Pharmacy, Linping Branch, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310000, P.R. China, Department of Pharmacology, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, Zhejiang 310000, P.R. China, Department of Pharmacology, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, Zhejiang 310000, P.R. China, Department of Pharmacy, Municipal Hospital Affiliated to Taizhou University, Taizhou, Zhejiang 318000, P.R. China
Published online on: February 16, 2024 https://doi.org/10.3892/ijmm.2024.5358
Article Number: 34
Copyright: © Fu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Calcium overload, a notable instigator of acute pancreatitis (AP), induces oxidative stress and an inflammatory cascade, subsequently activating both endogenous and exogenous apoptotic pathways. However, there is currently lack of available pharmaceutical interventions to alleviate AP by addressing calcium overload. In the present study, the potential clinical application of liposome nanoparticles (LNs) loaded with 1,2‑bis(2‑aminophenoxy)ethane‑N,N,N',N'‑tetraacetic acid tetrakis (acetoxymethyl ester) (BAPTA‑AM), a cell‑permeant calcium chelator, was investigated as a therapeutic approach for the management of AP. To establish the experimental models in vitro, AR42J cells were exposed to high glucose/sodium oleate (HGO) to induce necrosis, and in vivo, intra‑ductal taurocholate (TC) infusion was used to induce AP. The findings of the present study indicated that the use of BAPTA‑AM‑loaded LN (BLN) effectively and rapidly eliminated excessive Ca²⁺ and reactive oxygen species, suppressed mononuclear macrophage activation and the release of inflammatory cytokines, and mitigated pancreatic acinar cell apoptosis and necrosis induced by HGO. Furthermore, the systemic administration of BLN demonstrated promising therapeutic potential in the rat model of AP. Notably, BLN significantly enhanced the survival rates of rats subjected to the TC challenge, increasing from 37.5 to 75%. This improvement was attributed to the restoration of pancreatic function, as indicated by improved blood biochemistry indices and alleviation of pancreatic lesions. The potential therapeutic efficacy of BLN in rescuing patients with AP is likely attributed to its capacity to inhibit oxidative stress, prevent premature activation of zymogens and downregulate the expression of TNF‑α, IL‑6 and cathepsin B. Thus, BLN demonstrated promising value as a novel therapeutic approach for promptly alleviating the burden of intracellular Ca²⁺ overload in patients with AP.

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1	Chan YC and Leung PS: Acute pancreatitis: Animal models and recent advances in basic research. Pancreas. 34:1–14. 2007.
2	Jia A, Yang Z, Shi J, Liu J, Zhang K and Cui Y: MiR-325-3p alleviates acute pancreatitis via targeting RIPK3. Digest Dis Sci. 67:4471–4483. 2022.
3	Zhao Q, Zhang H, Huang J, Yu H, Li J, Che Q, Sun Y, Jin Y and Wu J: Melatonin attenuates the inflammatory response via inhibiting the C/EBP homologous protein-mediated pathway in taurocholate-induced acute pancreatitis. Int J Mol Med. 42:3513–3521. 2018.
4	Baron TH, DiMaio CJ, Wang AY and Morgan KA: American gastroenterological association clinical practice update: Management of pancreatic necrosis. Gastroenterology. 158:67–75. 2020.
5	Boxhoorn L, Voermans RP, Bouwense SA, Bruno MJ, Verdonk RC, Boermeester MA, van Santvoort HC and Besselink MG: Acute pancreatitis. Lancet. 396:7262020.
6	Heckler M, Hackert T, Hu K, Halloran CM, Büchler MW and Neoptolemos JP: Severe acute pancreatitis: Surgical indications and treatment. Langenbecks Arch Surg. 406:521–535. 2021.
7	Leppaniemi A, Tolonen M, Tarasconi A, Segovia-Lohse H, Gamberini E, Kirkpatrick AW, Ball CG, Parry N, Sartelli M, Wolbrink DRJ, et al: 2019 WSES guidelines for the management of severe acute pancreatitis. World J Emerg Surg. 14:272019.
8	Crockett SD, Wani S, Gardner TB, Falck-Ytter Y, Barkun AN, Crockett S, Falck-Ytter Y, Feuerstein J, Flamm S, Gellad Z, et al: American gastroenterological association institute guideline on initial management of acute pancreatitis. Gastroenterology. 154:1096–1101. 2018.
9	Portelli M and Jones CD: Severe acute pancreatitis: Pathogenesis, diagnosis and surgical management. Hepatob Pancreat Dis. 16:155–159. 2017.
10	Bang U, Semb S, Nojgaard C and Bendtsen F: Pharmacological approach to acute pancreatitis. World J Gastroenterol. 14:2968–2976. 2008.
11	Moggia E, Koti R, Belgaumkar AP, Fazio F, Pereira SP, Davidson BR and Gurusamy KS: Pharmacological interventions for acute pancreatitis. Cochrane Database Syst Rev. 4:D113842017.
12	Ward JB, Jenkins SA, Sutton R and Petersen OH: Is an elevated concentration of acinar cytosolic free ionised calcium the trigger for acute pancreatitis? Lancet. 346:1016–1019. 1995.
13	Mithöfer K, Fernández-Del Castillo C, Frick TW, Lewandrowski KB, Rattner DW and Warshaw AL: Acute hypercalcemia causes acute pancreatitis and ectopic trypsinogen activation in the rat. Gastroenterology. 109:239–246. 1995.
14	Sakorafas GH and Tsiotou AG: Etiology and pathogenesis of acute pancreatitis: Current concepts. J Clin Gastroenterol. 30:343–356. 2000.
15	Yang AL and McNabb-Baltar J: Hypertriglyceridemia and acute pancreatitis. Pancreatology. 20:795–800. 2020.
16	Weber H, Huhns S, Luthen F, Jonas L and Schuff-Werner P: Calpain activation contributes to oxidative stress-induced pancreatic acinar cell injury. Biochem Pharmacol. 70:1241–1252. 2005.
17	Orrenius S, Ankarcrona M and Nicotera P: Mechanisms of calcium-related cell death. Adv Neurol. 71:1371996.
18	Orrenius S, Gogvadze V and Zhivotovsky B: Calcium and mitochondria in the regulation of cell death. Biochem Bioph Res Commun. 460:72–81. 2015.
19	Gerasimenko J, Peng S and Gerasimenko O: Role of acidic stores in secretory epithelia. Cell Calcium. 55:346–354. 2014.
20	Xiao J, Lin H, Liu B and Jin J: CaMKII/proteasome/cytosolic calcium/cathepsin B axis was present in tryspin activation induced by nicardipine. Biosci Rep. 39:BSR201905162019.
21	Xie Z, Zhao M, Yan C, Kong W, Lan F, Narengaowa, Zhao S, Yang Q, Bai Z, Qing H and Ni J: Cathepsin B in programmed cell death machinery: Mechanisms of execution and regulatory pathways. Cell Death Dis. 14:2552023.
22	Reinheckel T, Deussing J, Roth W and Peters C: Towards specific functions of lysosomal cysteine peptidases: Phenotypes of mice deficient for cathepsin B or cathepsin L. Biol Chem. 382:7352001.
23	Sendler M, Dummer A, Weiss FU, Krüger B, Wartmann T, Scharffetter-Kochanek K, van Rooijen N, Malla SR, Aghdassi A, Halangk W, et al: Tumour necrosis factor α secretion induces protease activation and acinar cell necrosis in acute experimental pancreatitis in mice. Gut. 62:430–439. 2013.
24	Dawra R, Sah RP, Dudeja V, Rishi L, Talukdar R, Garg P and Saluja AK: Intra-acinar Trypsinogen activation mediates early stages of pancreatic injury but not inflammation in mice with acute pancreatitis. Gastroenterology. 141:2210–2217. 2011.
25	Shen Y, Malik SA, Amir M, Kumar P, Cingolani F, Wen J, Liu Y, Zhao E, Farris AB, Raeman R, et al: Decreased hepatocyte autophagy leads to synergistic IL-1β and TNF mouse liver injury and inflammation. Hepatology. 72:595–608. 2020.
26	Wu H, Zhang M, Li W, Zhu S and Zhang D: Stachydrine attenuates IL-1β-induced inflammatory response in osteoarthritis chondrocytes through the NF-κB signaling pathway. Chem Biol Interact. 326:1091362020.
27	Criddle DN: Reactive oxygen species, Ca(2+) stores and acute pancreatitis; a step closer to therapy? Cell Calcium. 60:180–189. 2016.
28	Wang Y, Pu M, Yan J, Zhang J, Wei H, Yu L, Yan X and He Z: 1,2-Bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic Acid Acetoxymethyl ester loaded reactive oxygen species responsive hyaluronic acid-bilirubin nanoparticles for acute kidney injury therapy via alleviating calcium overload mediated endoplasmic reticulum stress. ACS Nano. 17:472–491. 2023.
29	Quarato G, Llambi F, Guy CS, Min J, Actis M, Sun H, Narina S, Pruett-Miller SM, Peng J, Rankovic Z and Green DR: Ca²⁺-mediated mitochondrial inner membrane permeabilization induces cell death independently of Bax and Bak. Cell Death Differ. 29:1318–1334. 2022.
30	Tang Q, Jin M, Xiang J, Dong M, Sun H, Lau C and Li G: The membrane permeable calcium chelator BAPTA-AM directly blocks human ether a-go-go-related gene potassium channels stably expressed in HEK 293 cells. Biochem Pharmacol. 74:1596–1607. 2007.
31	Fu Z, Fan Q, Zhou Y, Zhao Y and He Z: Elimination of intracellular calcium overload by BAPTA-AM-loaded liposomes: A promising therapeutic agent for acute liver failure. ACS Appl Mater Interfaces. 11:39574–39585. 2019.
32	He Z, Tang H, You X, Huang K, Dhinakar A, Kang Y, Yu Q and Wu J: BAPTA-AM nanoparticle for the curing of acute kidney injury induced by ischemia/reperfusion. J Biomed Nanotechnol. 14:868–883. 2018.
33	Hong X, Wang H, Yang J, Lin B, Min Q, Liang Y, Huang P, Zhong Z, Guo S, Huang B and Xu YF: Systemic injury caused by taurocholate-induced severe acute pancreatitis in rats. Exp Ther Med. 24:4682022.
34	Laukkarinen JM, Van Acker GJ, Weiss ER, Steer ML and Perides G: A mouse model of acute biliary pancreatitis induced by retrograde pancreatic duct infusion of Na-taurocholate. Gut. 56:1590–1598. 2007.
35	Aho HJ, Suonpää K, Ahola RA and Nevalainen TJ: Experimental pancreatitis in the rat. Ductal factors in sodium taurocholate-induced acute pancreatitis. Exp Pathol. 25:731984.
36	Lange JF, van Gool J and Tytgat GNJ: Experimental pancreatitis in the rat: Role of bile reflux in sodium taurocholate-induced acute haemorrhagic pancreatitis. Eur Surg Res. 18:369–374. 1986.
37	Tani S, Otsuki M, Itoh H, Nakamura T, Fujii M, Okabayashi Y, Fujisawa T and Baba S: The protective effect of the trypsin inhibitor urinastatin on cerulein-induced acute pancreatitis in rats. Pancreas. 3:471–476. 1988.
38	Wang GP, Wen JM, Wen PM, Wilbur RRM, Zhou SMP and Xiao XP: The effect of Somatostatin, Ulinastatin and Salvia miltiorrhiza on severe acute pancreatitis treatment. Am J Med Sci. 346:371–376. 2013.
39	Sag S, Imamoglu M, Sarihan H, Yulug E, Alver A, Geze Saatci S and Cay A: Effects of carbon dioxide pneumoperitoneum on exocrine and endocrine functions, and oxidative state of rat pancreas. Biotech Histochem. 96:257–262. 2021.
40	Wenhong D, Jia Y, Weixing W, Xiaoyan C, Chen C, Sheng X, Hao J and Huang RS: Zerumbone Attenuates the severity of acute necrotizing pancreatitis and pancreatitis-induced hepatic injury. Mediat Inflamm. 2012:156507–156512. 2012.
41	Otani T and Matsukura A: Premature trypsinogen activation during cerulein pancreatitis in rats occurs inside pancreatic acinar cells. Pancreas. 20:421–422. 2000.
42	Saluja A, Dudeja V, Dawra R and Sah RP: Early Intra-acinar events in pathogenesis of pancreatitis. Gastroenterology. 156:1979–1993. 2019.
43	Mort JS and Buttle DJ: Cathepsin B. Int J Biochem Cell Biol. 29:715–720. 1997.
44	Winer J, Jung CK, Shackel I and Williams PM: Development and validation of real-time quantitative reverse transcriptasepolymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Anal Biochem. 270:41–49. 1999.
45	Gerasimenko JV, Gerasimenko OV and Petersen OH: The role of Ca2+ in the pathophysiology of pancreatitis. J Physiol. 592:269–280. 2014.
46	Maléth J and Hegyi P: Ca2+ toxicity and mitochondrial damage in acute pancreatitis: Translational overview. Philos Trans R Soc Lond B Biol Sci. 371:201504252016.
47	Ismail OZ and Bhayana V: Lipase or amylase for the diagnosis of acute pancreatitis? Clin Biochem. 50:1275–1280. 2017.
48	Szatmary P, Grammatikopoulos T, Cai W, Huang W, Mukherjee R, Halloran C, Beyer G and Sutton R: Acute pancreatitis: Diagnosis and treatment. Drugs. 82:1251–1276. 2022.
49	Das D, Mukherjee S, Das AS, Mukherjee M and Mitra C: Aqueous extract of black tea (Camellia sinensis) prevents ethanol+cholecystokinin-induced pancreatitis in a rat model. Life Sci. 78:2194–2203. 2006.
50	Guicciardi ME, Deussing J, Miyoshi H, Bronk SF, Svingen PA, Peters C, Kaufmann SH and Gores GJ: Cathepsin B contributes to TNF-alpha-mediated hepatocyte apoptosis by promoting mitochondrial release of cytochrome c. J Clin Invest. 106:1127–1137. 2000.
51	He R, Wang Z, Dong S, Chen Z and Zhou W: Understanding Necroptosis in pancreatic diseases. Biomolecules. 12:8282022.
52	Sendler M, Maertin S, John D, Persike M, Weiss FU, Krüger B, Wartmann T, Wagh P, Halangk W, Schaschke N, et al: Cathepsin B activity initiates apoptosis via digestive protease activation in pancreatic acinar cells and experimental pancreatitis. J Biol Chem. 291:14717–14731. 2016.
53	Zhang X, Lin Q and Zhou Y: Progress of study on the relationship between mediators of inflammation and apoptosis in acute pancreatitis. Digest Dis Sci. 52:1199–1205. 2007.
54	Lankisch PG, Apte M and Banks PA: Acute pancreatitis. Lancet. 386:85–96. 2015.
55	Song Y, Zhang Z, Yu Z, Xia G, Wang Y, Wang L, Peng C, Jiang B and Liu S: Wip1 Aggravates the Cerulein-induced cell autophagy and inflammatory injury by targeting STING/TBK1/IRF3 in acute pancreatitis. Inflammation. 44:1175–1183. 2021.
56	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.
57	Zhou X, Chen H, Wei X, He Y, Xu C and Weng Z: Establishment of a mouse severe acute pancreatitis model using retrograde injection of sodium taurocholate into the Biliopancreatic duct. J Vis Exp. 182:e631292022.
58	Petersen OH: Ca2+-induced pancreatic cell death: Roles of the endoplasmic reticulum, zymogen granules, lysosomes and endosomes. J Gastroenterol Hepatol. 23(Suppl 1): S31–S36. 2008.
59	Li J, Zhou R, Zhang J and Li Z: Calcium signaling of pancreatic acinar cells in the pathogenesis of pancreatitis. World J Gastroenterol. 20:16146–16152. 2014.
60	Wang Q, Bai L, Luo S, Wang T, Yang F, Xia J, Wang H, Ma K, Liu M, Wu S, et al: TMEM16A Ca²⁺-activated Cl⁻ channel inhibition ameliorates acute pancreatitis via the IP3R/Ca2+/NFκB/IL-6 signaling pathway. J Adv Res. 23:25–35. 2020.
61	Yan J, Wang Y, Zhang J, Liu X, Yu L and He Z: Rapidly blocking the calcium Overload/ROS production feedback loop to alleviate acute kidney injury via microenvironment-responsive BAPTA-AM/BAC Co-delivery Nanosystem. Small. 19:e22069362023.
62	Fanczal J, Pallagi P, Görög M, Diszházi G, Almássy J, Madácsy T, Varga Á, Csernay Biró P, Katona X, Tóth E, et al: TRPM2-mediated extracellular Ca²⁺ entry promotes acinar cell necrosis in biliary acute pancreatitis. J Physiol. 598:1253–1270. 2020.
63	Ricardo Carvalho VP, Figueira da Silva J, Buzelin MA, Antônio da Silva Júnior C, Carvalho Dos Santos D, Montijo Diniz D, Binda NS, Borges MH, Senna Guimarães AL, Rita Pereira EM and Gomez MV: Calcium channels blockers toxins attenuate abdominal hyperalgesia and inflammatory response associated with the cerulein-induced acute pancreatitis in rats. Eur J Pharmacol. 891:1736722021.
64	Zhang P, Yin X, Wang X, Wang J, Na G and Ирина Павловна К: Paeonol protects against acute pancreatitis by Nrf2 and NF-κB pathways in mice. J Pharm Pharmacol. 74:1618–1628. 2022.
65	Mayerle J, Sendler M, Hegyi E, Beyer G, Lerch MM and Sahin-Tóth M: Genetics, cell biology, and pathophysiology of pancreatitis. Gastroenterology. 156:1951–1968. 2019.
66	Werneburg NW, Guicciardi ME, Bronk SF and Gores GJ: Tumor necrosis factor-alpha-associated lysosomal permeabilization is cathepsin B dependent. Am J Physiol Gastrointest Liver Physiol. 283:G947–G956. 2002.
67	Forsmark CE, Swaroop Vege S, Wilcox CM and Campion EW: Acute pancreatitis. N Engl J Med. 375:1972–1981. 2016.
68	Hines OJ and Pandol SJ: Management of severe acute pancreatitis. BMJ. 367:l62272019.
69	Li L, Tucker RW, Hennings H and Yuspa SH: Chelation of intracellular Ca2+ inhibits murine keratinocyte differentiation in vitro. J Cell Physiol. 163:105–114. 1995.
70	Tymianski M, Spigelman I, Zhang L, Carlen PL, Tator CH, Charlton MP and Wallace MC: Mechanism of action and persistence of Neuroprotection by Cell-permeant Ca2+ chelators. J Cereb Blood Flow Metab. 14:911–923. 1994.
71	Toronyi É, Hamar J, Perner F and Szende B: Prevention of apoptosis reperfusion renal injury by calcium channel blockers. Exp Toxicol Pathol. 51:209–212. 1999.
72	Jang M, Shin M, Cho Y, Baik H, Kim S, Hwang E and Kim C: 1,2-bis (2-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid (BAPTA-AM) inhibits caffeine-induced apoptosis in human neuroblastoma cells. Neurosci Lett. 358:189–192. 2004.
73	Hsu S, Jan C and Liang W: The investigation of the pyrethroid insecticide lambda-cyhalothrin (LCT)-affected Ca²⁺ homeostasis and -activated Ca2⁺-associated mitochondrial apoptotic pathway in normal human astrocytes: The evaluation of protective effects of BAPTA-AM (a selective Ca²⁺ chelator). Neurotoxicology. 69:97–107. 2018.
74	Frosali S, Leonini A, Ettorre A, Di Maio G, Nuti S, Tavarini S, Di Simplicio P and Di Stefano A: Role of intracellular calcium and S-glutathionylation in cell death induced by a mixture of isothiazolinones in HL60 cells. Biochim Biophys Acta. 1793:572–583. 2009.
75	Nathens AB, Curtis JR, Beale RJ, Cook DJ, Moreno RP, Romand J, Skerrett SJ, Stapleton RD, Ware LB and Waldmann CS: Management of the critically ill patient with severe acute pancreatitis. Crit Care Med. 32:2524–2536. 2004.
76	Maher MM, Lucey BC, Gervais DA and Mueller PR: Acute pancreatitis: The role of imaging and interventional radiology. Cardiovasc Inter Rad. 27:208–225. 2004.
77	Nordestgaard AG, Wilson SE and Williams RA: Correlation of serum amylase levels with pancreatic pathology and pancreatitis etiology. Pancreas. 3:159–161. 1988.
78	Moridani MY and Bromberg IL: Lipase and pancreatic amylase versus total amylase as biomarkers of pancreatitis: An analytical investigation. Clin Biochem. 36:31–33. 2003.
79	Song R, Yu D and Park J: Changes in gene expression of tumor necrosis factor alpha and interleukin 6 in a canine model of caerulein-induced pancreatitis. Can J Vet Res. 80:236–241. 2016.
80	Jiang CY, Wang W, Tang JX and Yuan ZR: The adipocytokine resistin stimulates the production of proinflammatory cytokines TNF-α and IL-6 in pancreatic acinar cells via NF-κB activation. J Endocrinol Invest. 36:986–992. 2013.
81	Fang S, Li P, Zhu C, Han X, Bao P and Guo W: Research progress of ulinastatin in the treatment of liver diseases. Int J Clin Exp Patho. 13:2720–2726. 2020.