Improvement of the functionality of pancreatic Langerhans islets via reduction of bacterial contamination and apoptosis using phenolic compounds

Document Type: Original Article


1 Toxicology and Diseases Group, The Institute of Pharmaceutical Sciences (TIPS), Tehran University of Medical Sciences, Tehran, Iran

2 Department of Drug and Food, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

3 Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran


Objective(s): During type-1 diabetes treating by pancreatic islet transplantation, increasing oxidative stress and microbial contaminations are the main reasons of transplantation failure. In this study, we evaluated anti-apoptotic, antioxidant and antimicrobial potentials of phenolic compounds called ellagic acid (EA) and silybin on rat pancreatic islets.
Materials and Methods: By doing MTT assay, effective concentrations of EA and silybin were determined as 1500 and 2100 μM, respectively. Then, ELISA methods, flow cytometry and MIC were done to investigate antioxidant, anti-apoptotic and antibacterial effects of those compounds, respectively.
Results: Results of FITC Annexin-V and PI staining via flow cytometry, and also caspase-3 and -9 activities performed that EA has anti-apoptotic effects on pancreatic cells. Both compounds significantly diminished reactive oxygen species, and enhanced antioxidant power and insulin secretion. Furthermore, the minimum inhibitory concentration test indicated that these two have antibacterial effects on both Gram-positive and Gram-negative bacteria which usually contaminate the pancreatic islets.
Conclusion: These findings support that use of EA and silybin can improve the function of islets which are used in transplantation, along with decreasing islets bacterial contamination.


Main Subjects

1. Yin N, Chen T, Yu Y, Han Y, Yan F, Zheng Z, Chen Z. Facile mechanical shaking method is an improved isolation approach for islet preparation and transplantation. Exp Ther Med 2016; 12:3658-3664.
2. Rahimifard M, Navaei-Nigjeh M, Mahroui N, Mirzaei S, Siahpoosh Z, Nili-Ahmadabadi A, et al. Improvement in the function of isolated rat pancreatic islets through reduction of oxidative stress using traditional Iranian medicine. Cell J 2014; 16:147-163.
3. Santos‐Buelga C, Scalbert A. Proanthocyanidins and tannin‐like compounds–nature, occurrence, dietary intake and effects on nutrition and health. J Sci Food Agri 2000; 80:1094-1117.
4. Gu L, Kelm MA, Hammerstone JF, Beecher G, Holden J, Haytowitz D, et al. Concentrations of proanthocyanidins in common foods and estimations of normal consumption. J Nutr 2004; 134:613-617.
5. Clifford MN, Scalbert A. Ellagitannins–nature, occurrence and dietary burden. J Sci Food Agric 2000; 80:1118-1125.
6. Goodwin EC, Atwood WJ, Dimaio D. High-throughput cell-based screen for chemicals that inhibit infection by simian virus 40 and human polyomaviruses. J Virol 2009; 83:5630-5639.
7. Nohynek LJ, Alakomi HL, Kähkönen MP, Heinonen M, Helander IM, Oksman-Caldentey KM, et al. Berry phenolics: antimicrobial properties and mechanisms of action against severe human pathogens. Nutr Cancer 2006; 54:18-32.
8. Beserra AM, Calegari PI, Souza MC, Dos Santos RA, Lima JC, Silva R.M, et al. Gastroprotective and ulcer-healing mechanisms of ellagic acid in experimental rats. J Agric Food Chem 2011; 59:6957-6965.
9. Iakovleva L, Ivakhnenko A, Buniatian N. The protective action of ellagic acid in experimental myocarditis. Eksp Klin Farmakol 1997; 61:32-34.
10. Kaur R, Sharma U, Singh B, Arora S. Antimutagenic and antioxidant characteristics of Chukrasia tabularis a Juss extracts. Int J Toxicol 2011; 30:21-34.
11. Iqbal  J, Zaib S, Farooq U, Khan A, Bibi I,  Suleman S. Antioxidant, antimicrobial, and free radical scavenging potential of aerial parts of Periploca aphylla and Ricinus communis. ISRN Pharmacology 2012; doi:  10.5402/2012/563267.
12. Dell’agli M, Parapini S, Basilico N, Verotta L, Taramelli D, Berry C,  et al. In vitro studies on the mechanism of action of two compounds with antiplasmodial activity: Ellagic acid and 3, 4, 5-trimethoxyphenyl (6′-O-galloyl)-β-D-glucopyranoside. Planta Med 2003; 069:162-164.
13. Yoon CH, Chung SJ, Lee SW, Park YB, Lee SK, Park MC. Gallic acid, a natural polyphenolic acid, induces apoptosis and inhibits proinflammatory gene expressions in rheumatoid arthritis fibroblast-like synoviocytes. Joint Bone Spine 2013; 80:274-279.
14. Forester SC, Choy YY, Waterhouse AL Oteiza PI. The anthocyanin metabolites gallic acid, 3‐O‐methylgallic acid, and 2, 4, 6‐trihydroxybenzaldehyde decrease human colon cancer cell viability by regulating pro‐oncogenic signals. Mol Carcinog 2014; 53:432-439.
15. Baeeri M, Mohammadi-Nejad S, Rahimifard M, Navaei-Nigjeh M, Moeini-Nodeh S, Khorasani R,  et al. Molecular and biochemical evidence on the protective role of ellagic acid and silybin against oxidative stress-induced cellular aging. Mol Cell Biochem 2018; 441:21-33.
16. Vattem D, Shetty K. Biological functionality of ellagic acid: a review. J Food Biochem 2005; 29:234-266.
17. Friling RS, Bensimon A, Tichauer Y, Daniel V. Xenobiotic-inducible expression of murine glutathione S-transferase Ya subunit gene is controlled by an electrophile-responsive element. Proc Natl Acad Sci 1990; 87:6258-6262.
18. Wiseman H, Halliwell B. Damage to DNA by reactive oxygen and nitrogen species: role in inflammatory disease and progression to cancer. Biochem J 1996; 313:17-29.
19. Bisen PS, Bundela SS, harma A. Ellagic Acid? chemopreventive role in oral cancer. J Cancer Sci Therap 2012; 4:23-30.
20. Weisburg JH, Schuck AG, Reiss SE, Wolf BJ, Fertel SR, Zuckerbraun HL, et al. Ellagic acid, a dietary polyphenol, selectively cytotoxic to HSC-2 oral carcinoma cells. Anticancer Res 2013; 33:1829-1836.
21. Li TM, Chen GW, Su CC, Lin JG, Yeh CC, Cheng KC,  et al. Ellagic acid induced p53/p21 expression, G1 arrest and apoptosis in human bladder cancer T24 cells. Anticancer Res 2005; 25:971-979.
22. Zhang HM, Zhao Li, Xu H, Chen WW, Tao L. Research progress on the anticarcinogenic actions and mechanisms of ellagic acid. Cancer Biol Med 2014; 11:92-100.
23. Saeidnia S, Abdollahi M. Antioxidants: Friends or foe in prevention or treatment of cancer: The debate of the century. Toxicol Appl Pharmacol 2013; 271:49-63.
24. Loguercio C, Festi D. Silybin and the liver: from basic research to clinical practice. World J Gastroenterol 2011; 17:2288-2301.
25. Gazak R, Walterova D, Kren V. Silybin and silymarin-new and emerging applications in medicine. Curr Med Chem 2007; 14:315-338.
26. Dashti-Khavidaki S, Shahbazi F, Khalili H, Lessan-Pezeshki M. Potential renoprotective effects of silymarin against nephrotoxic drugs: a review of literature. J Pharm Pharm Sci 2012, 15:112-123.
27. Dehmlow C, Erhard J, De Groot H. Inhibition of Kupffer cell functions as an explanation for the hepatoprotective properties of silibinin. Hepatology 1996; 23:749-754.
29. Yoo HG, Jungm SN, Hwang YS, Park JS, Kim MH, Jeong M,  et al. Involvement of NF-KB and caspases in silibinin-induced apoptosis of endothelial cells. Int J Mol Med 2004; 13:81-86.
30. Katiyar SK, Roy AM,  Baliga MS. Silymarin induces apoptosis primarily through a p53-dependent pathway involving Bcl-2/Bax, cytochrome c release, and caspase activation. Mol Cancer Ther 2005; 4:207-216.
31. Fallahzadeh MK, Dormanesh B, Sagheb MM, Roozbeh J, Vessal G, Pakfetrat M, et al. Effect of addition of silymarin to renin-angiotensin system inhibitors on proteinuria in type 2 diabetic patients with overt nephropathy: a randomized, double-blind, placebo-controlled trial. Am J Kidney Dis 2012; 60:896-903.
32. Vessal G, Akmali M, Najafi P, Moein MR, Sagheb MM. Silymarin and milk thistle extract may prevent the progression of diabetic nephropathy in streptozotocin-induced diabetic rats. Renal Failure 2010; 32:733-739.
33. Ylipaasto P, Klingel K, Lindberg AM, Otonkoski T, Kandolf R, Hovi T, et al. Enterovirus infection in human pancreatic islet cells, islet tropism in vivo and receptor involvement in cultured islet beta cells. Diabetologia 2004; 47:225-239.
34. Bucher P, Oberholzer J, Bosco D, Mathe Z, Toso C, Bühler LH, Berney T, et al. Microbial surveillance during human pancreatic islet isolation. Transplant Int 2005; 18;584-589.
35. Shah SMM, Khan FA, Shah SMH, Chishti KA, et al. Evaluation of phytochemicals and antimicrobial activity of white and blue capitulum and whole plant of Silybum marianum. World Appl Sci J 2011; 12:1139-1144.
36. Moeini-Nodeh S, Rahimifard M, Baeeri M, Abdollahi M. Functional improvement in rats’ pancreatic islets using magnesium oxide nanoparticles through antiapoptotic and antioxidant pathways. Biol Trace Elem Res 2017; 175:146-155.
37. Rahimifard M, Navaii-Nigjeh M, Nilli-Ahmadabadi A, Pourkhalili N, Baeeri M, Mohammadirad A , et al. On the benefit of pure glycyrrhizic acid on the function and metabolic activity of isolated pancreatic Langerhans islets in vitro. Asian J Anim Vet Adv 2012; 7:1212-1218.
38. Shoae-Hagh P, Rahimifard M, Navaei-Nigjeh M, Baeeri M, Gholami M, Mohammadirad A, et al. Zinc oxide nanoparticles reduce apoptosis and oxidative stress values in isolated rat pancreatic islets. Biol Trace Elem Res 2014; 162:262-269.
39. Maqbool F, Bahadar H, Niaz K, Baeeri M, Rahimifard M, Navaei-Nigjeh M, et al. Effects of methyl mercury on the activity and gene expression of mouse Langerhans islets and glucose metabolism. Food Chem Toxicol 2016; 93:119-128.
40. Bahadar H, Maqbool F, Mostafalou S, Baeeri M, Rahimifard M, Navaei-Nigjeh M, et al. Assessment of benzene induced oxidative impairment in rat isolated pancreatic islets and effect on insulin secretion. Environ Toxicol Pharmacol 2015; 39:1161-1169.
41. Jowzi N, Rahimifard M, Navaei-Nigjeh M, Baeeri M, Darvishi B, Rezvanfar MA, et al. Reduction of chlorpyrifos-induced toxicity in human lymphocytes by selected phosphodiesterase inhibitors. Pest Biochem Physiol 2016; 128:57-62.
42. Yousefbeyk F, Gohari AR, Hashemighahderijani Z, Ostad SN, Sourmaghi MHS, Amini M, et al. Bioactive terpenoids and flavonoids from Daucus littoralis Smith subsp. hyrcanicus Rech. f, an endemic species of Iran. Daru 2014; 22:1-6.
43. Benavente-Garcia O, Castillo J. Update on uses and properties of citrus flavonoids: new findings in anticancer, cardiovascular, and anti-inflammatory activity. J Agric Food Chem 2008; 56:6185-6205.
44. Jadhav R, Puchchakayala G. Hypoglycemic and antidiabetic activity of flavonoids: boswellic acid, ellagic acid, quercetin, rutin on streptozotocin-nicotinamide induced type 2 diabetic rats. Int J Pharm Pharm Sci 2012; 4:251-256.
45. Rahimifard M, Maqbool F, Moeini-Nodeh S, Niaz K, Abdollahi M, Braidy N, et al. Targeting the TLR4 signaling pathway by polyphenol: A novel therapeutic strategy for neuroinflammation. Ageing Res Rev 2017; 36:11-19.
46. El-Shitany NA, El-Bastawissy EA, El-Desoky K. Ellagic acid protects against carrageenan-induced acute inflammation through inhibition of nuclear factor kappa B, inducible cyclooxygenase and proinflammatory cytokines and enhancement of interleukin-10 via an antioxidant mechanism. Int Immunopharmacol 2014; 19:290-299.
47. Ghasemi-Niri SF, Maqbool F, Baeeri M, Gholami M , Abdollahi M. Phosalone-induced inflammation and oxidative stress in the colon: Evaluation and treatment. World J Gastroenterol 2016; 22:4999-5011.
48. Tyagi AK, Agarwal C, Singh RP, Shroyer KR, Glode LM,  Agarwal R. Silibinin down-regulates survivin protein and mRNA expression and causes caspases activation and apoptosis in human bladder transitional-cell papilloma RT4 cells. Biochem Biophys Res Commun 2003; 312:1178-1184.
49. Slavov E, Georgiev IP, Dzhelebov P, Kanelov I, Andonova M, Georgieva TM, et al. High-fat feeding and Staphylococcus intermedius infection impair beta cell function and insulin sensitivity in mongrel dogs. Vet Res Commun 2010; 34:205-215.
50. Walker FR, Owens J, Ali S, Hodgson DM. Individual differences in glucose homeostasis: do our early life interactions with bacteria matter? Brain Behav Immun 2006; 20:401-409.
51. Alves MJ, Ferreira IC, Froufe HJ, Abreu RMV, Martins A ,Pintado M. Antimicrobial activity of phenolic compounds identified in wild mushrooms, SAR analysis and docking studies. J Appl Microbiol 2014; 115:346-357.
52. Nikolić DM, Đorđević PB, Dimitrijević-Srećković V, Džingalašević M, Belij S,  Kalezić N. Influence of the purification of human adult pancreatic islets on insulin secretion. Vojnosanit Pregl 2010; 67:128-131.
53. Mandel TE. Fetal islet transplantation in diabetic mice: a model for human islet transplants. Fetal Islet Transplantation 1988; 165-184.
54. Berger MG, Majumder K, Hodges JS, Bellin MD, Schwarzenberg SJ, Gupta S, et al. Microbial contamination of transplant solutions during pancreatic islet autotransplants is not associated with clinical infection in a pediatric population. Pancreatology 2016; 4:555-562.
55. Sugiura T, Mizuno T, Okamura Y, Ito T, Yamamoto Y,  Kawamura I, et al. Impact of bacterial contamination of the abdominal cavity during pancreaticoduodenectomy on surgical‐site infection. Br J Surg 2015; 102:1561-1566.
56. Baumann CA, Saltiel AR. Spatial compartmentalization of signal transduction in insulin action. Bioessays 2001; 23:215-222.
57. Fernández-Real JM, López-Bermejo A, Vendrell J, Ferri MJ, Recasens M, Ricart W. Burden of infection and insulin resistance in healthy middle-aged men. Diabetes Care 2006; 29:1058-1064.
58. Mohseni Salehi Monfared SS, Larijani B, Abdollahi M. Islet transplantation and antioxidant management: a comprehensive review. World J Gastroenterol 2009; 15:1153-61.