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Licensed Unlicensed Requires Authentication Published by De Gruyter October 18, 2018

Effect of Morus alba root bark extract on gene-level expression of inflammatory markers in rats subjected to ethanol and cerulein induced pancreatitis– influence of heat shock protein 70

  • Kavitha Yuvaraj and Arumugam Geetha EMAIL logo

Abstract

Background

Chronic pancreatitis (CP) is a persistent inflammation of the pancreas clinically presented with severe abdominal pain, progressive fibrosis, and loss of exocrine and endocrine functions. Inflammasomes, cytosolic multiprotein complexes which regulate the formation of proinflammatory cytokines, are influenced by various factors including heat shock proteins (HSPs). Morus alba L., or white mulberry root bark is a valued traditional Asian medicine with a diverse array of phytochemicals. The aim of this investigation was to define the modulatory action of methanolic extract of Morus alba root bark (MEMARB) on NLRP3 inflammasome, and HSPs in pancreas subjected to inflammatory insult.

Methods

Pancreatitis was induced in male albino Wistar rats by ethanol (0–36%) and cerulein (20 µg/kg b.wt., i.p.) for 5 weeks with or without MEMARB administration. Serum lipase/amylase (L/A) ratio, oxidative stress index (OSI) and reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio in the pancreas were evaluated. Levels of serum HSP70 was quantified by ELISA. NF-kappa B, NLRP3-ASC, caspase-1, IL-1β, IL-18, and HSP70 gene expression was quantified by quantitative real-time polymerase chain reaction (qPCR).

Results

L/A ratio and oxidative stress determined in terms of OSI and GSH/GSSG ratio were elevated in pancreatitis-induced rats. The levels were restored in MEMARB co-administered animals. Serum level of HSP70 was increased in pancreatitis-induced animals and dropped significantly in MEMARB co-administrated rats. Pancreatitis-induced group showed increased expression of NF-kappa B, IL-1β, IL-18, caspase-1, NLRP3-ASC and HSP70 mRNA than in MEMARB treated group.

Conclusions

It can be concluded that the M. alba root extract modulates the expression of HSP70 and NLRP3-ASC which might be attributed to its pancreato-protective effect.

Acknowledgements

The authors sincerely thank Dr Pramod K. Srivastava, Professor of Immunology and Medicine, and Director, Carole and Ray Neag Comprehensive Cancer Center, UConn School of Medicine, Farmington, Connecticut, U.S.A., for his kind gift of the HSP70 ELISA kit.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

[1] Whitcomb DC, Frulloni L, Garg P, Greer JB, Schneider A, Yadav D, et al. Chronic pancreatitis: an international draft consensus proposal for a new mechanistic definition. Pancreatology. 2016;16:218–24.10.1016/j.pan.2016.02.001Search in Google Scholar PubMed

[2] Kolodecik T, Shugrue C, Ashat M, Thrower EC. Risk factors for pancreatic cancer: underlying mechanisms and potential targets. Front Physiol. 2014;4:415.10.3389/fphys.2013.00415Search in Google Scholar PubMed

[3] Majumder S, Chari ST. Chronic pancreatitis. Lancet. 2016;387:1957–66.10.1016/S0140-6736(16)00097-0Search in Google Scholar PubMed

[4] Van Acker GJ, Weiss E, Steer ML, Perides G. Cause-effect relationships between zymogen activation and other early events in secretagogue-induced acute pancreatitis. Am J Physiol Gastrointest Liver Physiol. 2007;292:G1738–46.10.1152/ajpgi.00543.2006Search in Google Scholar PubMed

[5] Talukdar R, Sareen A, Zhu H, Yuan Z, Dixit A, Cheema H, et al. Release of cathepsin b in cytosol causes cell death in acute pancreatitis. Gastroenterology. 2016;151:747–58.10.1053/j.gastro.2016.06.042Search in Google Scholar PubMed PubMed Central

[6] Dawra R, Sah RP, Dudeja V, Rishi L, Talukdar R, Garg P, et al. Intra-acinar trypsinogen activation mediates early stages of pancreatic injury but not inflammation in mice with acute pancreatitis. Gastroenterology. 2011;141:2210–17.10.1053/j.gastro.2011.08.033Search in Google Scholar PubMed PubMed Central

[7] Hoque R, Malik A, Gorelick F, Mehal W. Sterile inflammatory response in acute pancreatitis. Pancreas. 2012;41:353–57.10.1097/MPA.0b013e3182321500Search in Google Scholar PubMed PubMed Central

[8] Habtezion A. Inflammation in acute and chronic pancreatitis. Curr Opin Gastroenterol. 2015;31:395–99.10.1097/MOG.0000000000000195Search in Google Scholar PubMed PubMed Central

[9] Hoque R, Mehal WZ. Inflammasomes in pancreatic physiology and disease. Am J Physiol Gastrointest Liver Physiol. 2015;308:G643–51.10.1152/ajpgi.00388.2014Search in Google Scholar PubMed PubMed Central

[10] Abderrazak A, Syrovets T, Couchie D, El Hadri K, Friguet B, Simmet T, et al. NLRP3 inflammasome: from a danger signal sensor to a regulatory node of oxidative stress and inflammatory diseases. Redox Biol. 2015;4:296–307.10.1016/j.redox.2015.01.008Search in Google Scholar PubMed PubMed Central

[11] Dawra RK, Dudeja V, Saluja AK. Heat shock proteins as modulators of pancreatitis. Pancreapedia Exocrine Pancreas Knowl Base. 2016. DOI: 10.3998/panc.2016.18Search in Google Scholar

[12] Kollar P, Bárta T, Hošek J, Souček K, Závalová VM, Artinian S, et al. Prenylated flavonoids from Morus alba L. cause inhibition of G1/S transition in THP-1 human leukemia cells and prevent the lipopolysaccharide-induced inflammatory response. Evid Based Complement Alternat Med. 2013;2013:350519–31.10.1155/2013/350519Search in Google Scholar

[13] Fariss MW, Reed DJ. High-performance liquid chromatography of thiols and disulfides: dinitrophenol derivatives. Methods Enzymol. 1987;143:101–09.10.1016/0076-6879(87)43018-8Search in Google Scholar PubMed

[14] Harma M, Harma M, Erel O. Increased oxidative stress in patients with hydatidiform mole. Swiss Med Wkly. 2003;133:563–66.10.4414/smw.2003.10397Search in Google Scholar PubMed

[15] Miller NJ, Rice-Evans C, Davies MJ, Gopinathan V, Milner A. A novel method for measuring antioxidant capacity and its application to monitoring the antioxidant status in premature neonates. Clin Sci. 1993;84:407–12.10.1042/cs0840407Search in Google Scholar PubMed

[16] Wagner H, Bauer R, Melchart D, Xiao PG, Staudinger A. Chromatographic fingerprint analysis of herbal medicines. Berlin, Germany: Springer, 2011.10.1007/978-3-7091-0763-8Search in Google Scholar

[17] Venkatesh Kumar R, Chauhan S. Mulberry: life enhancer. J Med Plant Res. 2008;2:271–78.Search in Google Scholar

[18] Vonlaufen A, Wilson JS, Pirola RC, Apte MV. Role of alcohol metabolism in chronic pancreatitis. Alcohol Res Health. 2007;30:48–54.Search in Google Scholar PubMed

[19] Deng X, Wang L, Elm MS, Gabazadeh D, Diorio GJ, Eagon PK, et al. Chronic alcohol consumption accelerates fibrosis in response to cerulein-induced pancreatitis in rats. Am J Pathol. 2005;166:93–106.10.1016/S0002-9440(10)62235-3Search in Google Scholar PubMed

[20] Kim H. Cerulein pancreatitis: oxidativestress, inflammation, and apoptosis. Gut Liver. 2008;2:74–80.10.5009/gnl.2008.2.2.74Search in Google Scholar PubMed PubMed Central

[21] Frulloni L, Patrizi F, Bernardoni L, Cavallini G. Pancreatic hyperenzymemia: clinical significance and diagnostic approach. JOP. 2005;6:536–41.Search in Google Scholar PubMed

[22] Devanath A, Kumari J, Joe J, Peter S, Rajan S, Sabu L, et al. Usefulness of lipase/amylase ratio in acute pancreatitis in South Indian population. Indian J Clin Biochem. 2009;24:361–65.10.1007/s12291-009-0065-3Search in Google Scholar PubMed PubMed Central

[23] Shahedi K, Pandol SJ, Hu R. Oxidative stress and alcoholic pancreatitis. J Gastroenterol Hepatol Res. 2013;2:335–42.Search in Google Scholar

[24] Wittel UA, Bachem M, Siech M. Oxygen radical production precedes alcohol-induced acute pancreatitis in rats. Pancreas. 2003;26:e74–80.10.1097/00006676-200305000-00018Search in Google Scholar PubMed

[25] Kang R, Lotze MT, Zeh HJ, Billiar TR, Tang D. Cell death and DAMPs in acute pancreatitis. Mol Med. 2014;20:466–77.10.2119/molmed.2014.00117Search in Google Scholar PubMed PubMed Central

[26] Rakonczay Z, Hegyi P, Takacs T, McCarroll J, Saluja AK. The role of NF-κB activation in the pathogenesis of acute pancreatitis. Gut. 2008;57:259–67.10.1136/gut.2007.124115Search in Google Scholar PubMed

[27] Chen X, Ji B, Han B, Ernst SA, Simeone D, Logsdon CD. NF-κB activation in pancreas induces pancreatic and systemic inflammatory response. Gastroenterology. 2002;122:448–57.10.1053/gast.2002.31060Search in Google Scholar PubMed

[28] Compan V, Martín-Sánchez F, Baroja-Mazo A, López-Castejón G, Gomez AI, Verkhratsky A, et al. Apoptosis-associated speck-like protein containing a CARD forms specks but does not activate caspase-1 in the absence of NLRP3 during macrophage swelling. J Immunol. 2015;194:1261–73.10.4049/jimmunol.1301676Search in Google Scholar PubMed PubMed Central

[29] HošEk J, Bartos M, Chudík S, Dall’Acqua S, Innocenti G, Kartal M, et al. Natural compound cudraflavone B shows promising anti-inflammatory properties in vitro. J Nat Prod. 2011;74:614–19.10.1021/np100638hSearch in Google Scholar PubMed

[30] Zelová H, Hanáková Z, ČErmáková Z, ŠMejkal K, Dalĺ Acqua S, Babula P, et al. Evaluation of anti-inflammatory activity of prenylated substances isolated from Morus albaMorus nigra. J Nat Prod. 2014;77:1297–303.10.1021/np401025fSearch in Google Scholar PubMed

[31] Ethridge RT, Ehlers RA, Hellmich MR, Rajaraman S, Evers BM. Acute pancreatitis results in induction of heat shock proteins 70 and 27 and heat shock factor-1. Pancreas. 2000;21:248–56.10.1097/00006676-200010000-00005Search in Google Scholar PubMed

[32] Hosokawa N, Hirayoshi K, Kudo H, Takechi H, Aoike A, Kawai K, et al. Inhibition of the activation of heat shock factor in vivo and in vitro by flavonoids. Mol Cell Biol. 1992;12:3490–98.10.1128/MCB.12.8.3490Search in Google Scholar PubMed

[33] Phillips PA, Dudeja V, McCarroll JA, Borja-Cacho D, Dawra RK, Grizzle WE, et al. Triptolide induces pancreatic cancer cell death via inhibition of heat shock protein 70. Cancer Res. 2007;67:9407–16.10.1158/0008-5472.CAN-07-1077Search in Google Scholar PubMed

[34] Sendler M, Mayerle J, Lerch MM. Necrosis, apoptosis, necroptosis, pyroptosis: it matters how acinar cells die during pancreatitis. Cell Mol Gastroenterol Hepatol. 2016;2:407–08.10.1016/j.jcmgh.2016.05.007Search in Google Scholar PubMed PubMed Central

[35] Schmitt E, Gehrmann M, Brunet M, Multhoff G, Garrido C. Intracellular and extracellular functions of heat shock proteins: repercussions in cancer therapy. J Leukoc Biol. 2007;81:15–27.10.1189/jlb.0306167Search in Google Scholar PubMed

Received: 2017-11-07
Accepted: 2018-07-17
Published Online: 2018-10-18

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