Skip to main content

Advertisement

SpringerLink
Log in

Effect of diosmin on apoptotic signaling molecules in N-nitrosodiethylamine-induced hepatocellular carcinoma in experimental rats

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The aim of the present study was to evaluate the antioxidant and chemopreventive efficiency of diosmin against N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis in adult male rats. Rats were classified into four groups as follows: Group I: Control, Group II: NDEA-induced hepatocellular carcinogenic rats, Group III: Cancer-bearing animals treated with diosmin (200 mg/kg/body weight/day) orally for 28 days, Group IV: Control animals treated with diosmin (200 mg/kg/body weight/day) alone for 28 days. The model of NDEA-induced HCC rats elicited significant increases in alpha-fetoprotein (AFP), lipid peroxidation (LPO) and increase in anti-apoptotic proteins (Bcl-2, Bcl-xL and Mcl-1) with a concomitant significant decline in liver antioxidant enzymes, pro-apoptotic (Bax and Bad) and caspase-3 &-9 proteins. The oral administration of diosmin as a protective agent normalized the altered levels of AFP, LPO, antioxidant enzymes, pro- and anti-apoptotic proteins as well as caspase-3 and -9 proteins. Transmission electron microscopical studies also revealed that treatment of diosmin has a perspective anti-cancer activity by rearranging hepatic cell structure and its integrity. Results of this study suggest that diosmin may be one of a pharmacological and therapeutic representative against hepatocellular carcinoma.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Mittal S, El-Serag HB (2013) Epidemiology of HCC: Consider the Population. J Clin Gastroenterol 47:S2-S6. https://doi.org/10.1097/MCG.0b013e3182872f29

    Article  PubMed Central  Google Scholar 

  2. Caillot F, Derambure C, Bioulac-Sage P, Francois A, Goria O, Hiron M, Daveau M, Salier JP (2009) Transient and etiology-related transcription regulation in cirrhosis prior to hepatocellular carcinoma occurrence. World J Gastroenterol 15:300–309. https://doi.org/10.3748/wjg.15.300

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Gurtsevitch VE (2008) Human oncogenic viruses: hepatitis B and hepatitis C viruses and their role in hepatocarcinogenesis. Biochem (Mosc) 73:504–513. https://doi.org/10.1134/S0006297908050039

    Article  CAS  Google Scholar 

  4. Lewinska A, Siwak J, Rzeszutek I, Wnuk M (2015) Diosmin induces genotoxicity and apoptosis in DU145 prostate cancer cell line. Toxicol In Vitro 29:417–425. https://doi.org/10.1016/j.tiv.2014.12.005

    Article  CAS  PubMed  Google Scholar 

  5. Bouskela E, Cyrino FZ, Lerond L (1997) Summary of Product Characteristics. Daflon 500 mg film-coated tablets. Micronized purified flavonoidfraction. Br J Pharmacol 122:1611–1616

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Firuzi O, Miri R, Tavakkoli M, Saso L (2011) Antioxidant therapy: current status and future prospects. Curr Med Chem 18:3871–3888. https://doi.org/10.2174/092986711803414368

    Article  CAS  PubMed  Google Scholar 

  7. Silvestro L, Tarcomnicu I, Dulea C, Attili NR, Ciuca V, Peru D, Savu SR (2013) Confirmation of diosmetin 3-O-glucuronide as major metabolite of diosmin in humans, using micro-liquidchromatography-mass spectrometry and ion mobility mass spectrometry. Anal Bioanal Chem 405:8295–8310. https://doi.org/10.1007/s00216-013-7237-y

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Tanaka T, Makita H, Kawabata K, Mori H, Kakumoto M, Satoh K, Hara A, Sumida T, Tanaka T, Ogawa H (1997) Chemoprevention of azoxymethane-induced rat colon carcinogenesis by the naturally occurring flavonoids, diosmin and hesperidin. Carcinogenesis 18:957–965

    Article  CAS  PubMed  Google Scholar 

  9. Katsenis K (2005) Micronized purified flavonoid fraction (MPFF): a review of its pharmacological effects, therapeutic efficacy and benefits in the management of chronic venous insufficiency. Curr Vasc Pharmacol 3:1–9. https://doi.org/10.2174/1570161052773870

    Article  CAS  PubMed  Google Scholar 

  10. Bogucka-Kocka A, Woźniak M, Feldo M, Kockic J, Szewczyk K (2013) Diosmin-isolation techniques, determination in plant material and pharmaceutical formulations, and clinical use. Nat Prod Commun 8:545–550

    CAS  PubMed  Google Scholar 

  11. No Author (2004) Diosmin monograph. Altern Med Rev 9:308–311

  12. Lyseng-Williamson KA, Perry CM (2003) Micronised purified flavonoid fraction: a review of its use in chronic venous insufficiency, venous ulcers and haemorrhoids. Drugs 63:71–100

    Article  CAS  PubMed  Google Scholar 

  13. Srinivasan S, Pari L (2012) Ameliorative effect of diosmin, a citrus flavonoid against streptozotocin-nicotinamide generated oxidative stress induced diabetic rats. Chem Biol Interact 195:43–51. https://doi.org/10.1016/j.cbi.2011.10.003

    Article  CAS  PubMed  Google Scholar 

  14. Rehman MU, Tahir M, Quaiyoom Khan A, Khan R, Lateef A, Hamiza OO (2013) Diosmin protects against trichloroethylene-induced renal injury in Wistar rats: plausible role of p53, Bax and caspases. Br J Nutr 110:699–710. https://doi.org/10.1017/S0007114512005752

    Article  CAS  PubMed  Google Scholar 

  15. Tahir M, Rehman MU, Lateef A, Khan R, Khan AQ, Qamar W (2013) Diosmin protects againstethanol-induced hepatic injury via alleviation of inflammation and regulation of TNF- alpha and NF-kappa B activation. Alcohol 47:131–139. https://doi.org/10.1016/j.alcohol.12.010

    Article  CAS  PubMed  Google Scholar 

  16. Queenthy SS, John B (2013) Diosmin exhibits anti-hyperlipidemic effects in isoproterenol induced myocardial infarcted rats. Eur J Pharmacol 718:213–218. https://doi.org/10.1016/j.ejphar.2013.08.031

    Article  CAS  PubMed  Google Scholar 

  17. Tahir M, Rehman MU, Lateef A, Khan AQ, Khan R, Qamar W (2013) Diosmin abrogates chemically induced hepatocarcinogenesis via alleviation of oxidative stress, hyperproliferative and inflammatory markers in murine model. Toxic Lett 220:205–218. https://doi.org/10.1016/j.toxlet.2013.04.004

    Article  CAS  Google Scholar 

  18. Lewinska A, Siwak J, Rzeszutek I, Wnuk M (2015) Diosmin induces genotoxicity and apoptosis in DU145 prostate cancer cell line. ToxicolIn Vitro 29:417–425. https://doi.org/10.1016/j.tiv.2014.12.005

    Article  CAS  Google Scholar 

  19. Arab HH, Salama SA, Omar HA, Arafa ES, Maghrabi IA (2015) Diosmin protects against ethanol- induced gastric injury in rats: novel anti-ulcer actions. PLoS ONE 10:e0122417. https://doi.org/10.1371/journal.pone.0122417

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Perumal S, HariBabu L, Langeswaran K, Gowthamkumar S, Balasubramanian MP (2015) Pharmacological potential of natural flavonoid diosmin against n-nitrosodiethylamine- induced hepatocellular carcinogenesis in wistar albino rats. Int J Res Biosci 4:25–36

    Google Scholar 

  21. Hogberg J, Larson RE, Kristoferson A, Orrenius S (1974) NADPH-dependent reductase solubilised from microsomes by peroxidation and its activity. Biochem Biophys Res Commun 56:836–842. https://doi.org/10.1016/0006-291X(74)90681-0

    Article  CAS  PubMed  Google Scholar 

  22. Devasagayam TPA, Tarachand U (1987) Decreased lipid peroxidation in the rat kidney during gestation. Biochem Biophys Res Commun 145:134–138. https://doi.org/10.1016/0006-291X(87)91297-6

    Article  CAS  PubMed  Google Scholar 

  23. Marklund G, Marklund (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474. https://doi.org/10.1111/j.1432-1033.1974.tb03714.x

    Article  CAS  PubMed  Google Scholar 

  24. Sinha AK (1972) Colorimetric assay of catalase. Anal Biochem 47(2):389–394. https://doi.org/10.1016/0003-2697(72)90132-7

    Article  CAS  PubMed  Google Scholar 

  25. Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hekstra WG (1973) Selenium biochemical role as a component of glutathione peroxidase purification and assay. Science 179:588–590. https://doi.org/10.1126/science.179.4073.588

    Article  CAS  PubMed  Google Scholar 

  26. Moron MS, Depierre JW, Mannervik B (1979) Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochem Biophys Acta 582:67–78. https://doi.org/10.1016/0304-4165(79)90289-7

    Article  CAS  PubMed  Google Scholar 

  27. Omaye ST, Turnbull JD, Sauberlich HE (1979) Selected methods for the determination of ascorbic acid in animal cells, tissues and fluids. Methods Enzymol 62:3–11. https://doi.org/10.1016/0076-6879(79)62181-X

    Article  CAS  PubMed  Google Scholar 

  28. Desai ID (1984) Vitamin E analysis methods for animal tissues. Methods Enzymol 105:138–147

    Article  CAS  PubMed  Google Scholar 

  29. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    CAS  Google Scholar 

  30. Robinson G, Gray T (1996) Electron microscopy 2: practical procedures. In: Bancroft JD, Stevens A (eds) Theory and practice of histological techniques, 4th edn. Churchill Livingstone, Edinburgh, pp. 586

    Google Scholar 

  31. Dimitrios S, Grigoris A, Antonios M, Dimitrios K (2012) Chemoprevention of liver cancer by plant polyphenols. Food ChemToxicol 50:2155–2170. https://doi.org/10.1016/j.fct.2012.04.002

    Article  CAS  Google Scholar 

  32. Glauert HP, Calfee-Mason K, Stemm DN, Tharappel JC, Spear BT (2010) Dietary antioxidants in the prevention of hepatocarcinogenesis: a review. Nutr Food Res 54:875–896. https://doi.org/10.1002/mnfr.200900482

    Article  CAS  Google Scholar 

  33. Shahjahan M, Vani G, Shyamaladevi CS (2005) Effect of Solanumtrilobatum on theantioxidant status during diethyl nitrosamine induced and phenobarbital promoted hepatocarcinogenesis in rat. Chem Biol Interact 156:113–123

    Article  CAS  PubMed  Google Scholar 

  34. Yang CS, Lambert JD, Ju J, Gang Lu, Sang S (2007) Tea and cancer prevention: molecular mechanisms and human relevance. Toxicol Appl Pharmacol 224:265–273. https://doi.org/10.1016/j.taap.2006.11.024

    Article  CAS  PubMed  Google Scholar 

  35. Plochmann K, Korte G, Koutsilieri E, Richling E, Riederer P, Rethwilm A, Schreier P, Scheller C (2007) Structure activity relationships of flavonoid-induced cytotoxicity on human leukemia cells. Arch Biochem Biophys 460:1–9. https://doi.org/10.1016/j.abb.2007.02.003

    Article  CAS  PubMed  Google Scholar 

  36. Sell S (2008) Alpha-fetoprotein, stem cells and cancer: how study of the production of alpha- fetoprotein during chemical hepatocarcinogenesis led to reaffirmation of the stem cell theory of cancer. Tumour Biol 29:161–180. https://doi.org/10.1159/000143402

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Singhal A, Jayaraman M, Dhanasekaran DN, Kohli V (2012) Molecular and serum markers in hepatocellular carcinoma: predictive tools for prognosis and recurrence. Crit Rev Oncol Hematol 82:116–140. https://doi.org/10.1016/j.critrevonc.2011.05.005

    Article  PubMed  Google Scholar 

  38. Shabnam M, Shilpa B, Naveen C, Deepshikha K, Jain SK (2012) Den+ 2-AAF-induced multistep hepatotumorigenesis in Wistarrats:suppoertie evidence and insights. Protoplasma 250:175–183. https://doi.org/10.1007/s00709-012-0392-8

    Article  CAS  Google Scholar 

  39. Nickerson J (2001) Experimental observations of a nuclear matrix. J Cell Sci 4:463–474

    Google Scholar 

  40. Rajkapoor B, Murugesh N, Chodon D, Sakthisekaran D (2005) Chemoprevention of N-nitrosodiethylamine induced phenobarbitol promoted liver tumors in rat by extract of Indigofera aspalathoides. Biol Pharm Bull 28:364–366. https://doi.org/10.1248/bpb.28.364

    Article  CAS  PubMed  Google Scholar 

  41. Lansky EP, Newman RA (2007) Punicagranatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. J Ethnopharmacol 109:177–206. https://doi.org/10.1016/j.jep.2006.09.006

    Article  CAS  PubMed  Google Scholar 

  42. Subash S, Essa MM, Al-Asmi A, Al-Adawi S, Vaishnav R, Braidy N (2014) Pomegranate from Oman alleviates the brain oxidative damage in transgenic mouse model of Alzheimer’s disease. J Tradit Complement Med 4:232–238. https://doi.org/10.4103/2225-4110.139107

    Article  PubMed  PubMed Central  Google Scholar 

  43. Ahmed S, Mundhe N, Borgohain M, Chowdhury L, Kwatra M, Bolshette N, Ahmed A, Lahkar M (2016) Diosmin modulates the NF-kB signal transduction pathways and downregulation of various oxidative stress markers in alloxan-induced diabetic nephropathy. Inflammation 39:1783–1797. https://doi.org/10.1007/s10753-016-0413-4

    Article  CAS  PubMed  Google Scholar 

  44. Shanmugam KR, Mallikarjuna K, Nishanth K, Kuo CH, Sathyavelu Reddy K (2011) Protective effect of dietary ginger on antioxidant enzymes and oxidative damage in experimental diabetic rat tissues. Food Chem 124:1436–1442

    Article  CAS  Google Scholar 

  45. Zaanan A, Williet N, Hebbar M, Dabakuyo TS, Fartoux L (2013) Gemcitabine plus oxaliplatin in advanced hepatocellular carcinoma: a large multicenter AGEO study. J Hepatol 58:81–88. https://doi.org/10.1016/j.jhep.2012.09.006

    Article  CAS  PubMed  Google Scholar 

  46. Lobo V, Patil A, Phatak A, Chandra N (2010) Free radicals, antioxidants and functional foods: impact on human health. Pharmacogn Rev 4:118–126. https://doi.org/10.4103/0973-7847.70902

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Devaki T, Binuclara J, Raghunandakumar S, Asokkumar S, Naveenkumar C, Premkumar T (2013) Anti-fibrotic effect of diosmin against dmn-induced liver fibrosisIn rats: a biochemical analysis. IJRPNS 2:283–292

    Google Scholar 

  48. Gönüllü U, Kato H (1995) Nuclear DNA and nuclear protein content of tumor cell in adenocarcinoma of the lung. Lung Cancer 13:13–19. https://doi.org/10.1016/0169-5002(95)00475G

    Article  PubMed  Google Scholar 

  49. Abdel-Daim MM, Khalifa HA, Abushouk AI, Dkhil MA, Al-Quraishy SA (2017) Diosmin attenuates methotrexate-induced hepatic, renal, and cardiac injury: a biochemical and histopathological study in mice. Oxid Med Cell Longev. https://doi.org/10.1155/2017/3281670

    Article  PubMed  PubMed Central  Google Scholar 

  50. Kang MH, Reynolds CP (2009) Bcl-2 inhibitors: targeting mitochondrial apoptotic pathways in cancer therapy. Clin Cancer Res 15:1126–1132. https://doi.org/10.1158/1078-0432.CCR-08-0144

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Professor and Head, Department of Pharmacology & Environmental Toxicology, University of Madras for providing Transmission Electron Microscopy (TEM) facility.

Author information

Authors and Affiliations

  1. Department of Biochemistry, School of Life Sciences, Vels University, Chennai, Tamil Nadu, India

    Srinivasan Perumal & M. P. Balasubramanian

  2. Department of Bioinformatics, Alagappa University, Karaikudi, Tamil Nadu, India

    Kulanthaivel Langeshwaran

  3. Central Research Laboratory, Meenakshi Ammal Dental College, Meenakshi Academy of Higher Education and Research (MAHER), Chennai, Tamil Nadu, India

    Jayaraman Selvaraj, Rajagopal Ponnulakshmi & B. Shyamaladevi

Authors
  1. Srinivasan Perumal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kulanthaivel Langeshwaran
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jayaraman Selvaraj
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rajagopal Ponnulakshmi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. B. Shyamaladevi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. P. Balasubramanian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jayaraman Selvaraj.

Ethics declarations

Conflict of interest

The authors declared that there is no conflict of interest from any of the authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Perumal, S., Langeshwaran, K., Selvaraj, J. et al. Effect of diosmin on apoptotic signaling molecules in N-nitrosodiethylamine-induced hepatocellular carcinoma in experimental rats. Mol Cell Biochem 449, 27–37 (2018). https://doi.org/10.1007/s11010-018-3339-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-018-3339-3

Keywords

Search

Navigation