Antiproliferative activity of Brassica nigra seeds extract in liver tissue of mice exposed to phenobarbital

Ibtehal Naseer Salman; Dalya Basil Hanna; Bahir Abdul-Razzaq Mshimesh

doi:10.32947/ajps.v22i1.826

Authors

Ibtehal Naseer Salman Department of Pharmacology/ College of Pharmacy/ Mustansiriyah University, Baghdad, Iraq
Dalya Basil Hanna Department of Clinical Laboratory Sciences/ College of Pharmacy/ Mustansiriyah University, Baghdad, Iraq
Bahir Abdul-Razzaq Mshimesh Department of Pharmacology/ College of Pharmacy/ Mustansiriyah University, Baghdad, Iraq

DOI:

https://doi.org/10.32947/ajps.v22i1.826

Keywords:

Hepatocellular proliferation, Phenobarbital, Brassica seeds

Abstract

Hepatocellular proliferation is one of the most common causes of hepatocellular carcinoma (HCC), a type of cancer that is widely distributed disease. Hepatocellular carcinoma treatment has numerous barriers, including ineffectiveness, side effects, and drug resistance to currently available

treatments. Previous studies showed that a high intake of Brassica vegetables has been associated to a decreased risk of a number of malignancies. The aim of this study is the evaluation of antiproliferative activity of Brassica nigra seeds extract in mice exposed to phenobarbital. Brassica nigra seeds where extracted; phytochemical analysis of the extract was done that including phytochemical screening tests and Gas chromatography-mass spectrometry (GC-MS) analysis. Antiproliferative activity of hydro alcoholic Brassica seeds extract has been studied by 800mg/kg and compare with control group (given normal saline), phenobarbital group (Phenobarbital 75mg/kg) and combination group (Brassica extract 800mg/kg+ Phenobarbital 75mg/kg). The GC-MS analysis revealed the presence of isothiocynate compound. Histologically phenobarbital induced severe hepatocellular proliferation (hyperplasia and hypertrophy), glass ground cytoplasm, while Brassica seeds extract produce improvement in histopathological changes that include mild scattered proliferation picture and eosinophilic cytoplasm. In comparison to phenobarbital group, Combination groups pretreated with Brassica nigra seeds for 14 days and phenobarbital for 7 days caused significant reduction relative liver weight and alanine aminotransferase (ALT) Brassica nigra seeds extract have isothiocynate as main compound it showed antiproliferative action on the liver tissue, implying that it may have a promising effect in minimizing the risk of liver cancer.

References

- Issues CC. Current Clinical Issues Liver Cancer on the Rise TO. 2005;1029–32.‏ DOI: https://doi.org/10.7326/0003-4819-142-12_Part_1-200506210-00024

- Maronpot RR. Biological basis of differential susceptibility to hepatocarcinogenesis among mouse strains. J Toxicol Pathol. 2009;22(1):11–33. DOI: https://doi.org/10.1293/tox.22.11

- Whysner J, Ross PM, Williams GM. Phenobarbital mechanistic data and risk assessment: Enzyme induction, enhanced cell proliferation, and tumor promotion. Pharmacol Ther. 1996;71(1–2):153–91. DOI: https://doi.org/10.1016/0163-7258(96)00067-8

- Müller PB, Taboada J, Hosgood G, Partington BP, VanSteenhouse JL, Taylor HW, et al. Effects of long-term phenobarbital treatment on the liver in dogs. J Vet Intern Med. 2000;14(2):165–71.

- Smith TK, Lund EK, Clarke RG, Bennett RN, Johnson IT. Effects of Brussels sprout juice on the cell cycle and adhesion of human colorectal carcinoma cells (HT29) in vitro. J Agric Food Chem. 2005;53(10):3895–901. DOI: https://doi.org/10.1021/jf048025v

- Van Poppel G, Verhoeven DTH, Verhagen H, Goldbohm RA. Brassica vegetables and cancer prevention: Epidemiology and mechanisms. Adv Exp Med Biol. 2000;472:159–68. DOI: https://doi.org/10.1007/978-1-4757-3230-6_14

- Giovannucci E, Rimm EB, Liu Y, Stampfer MJ, Willett WC. A Prospective Study of Cruciferous Vegetables and Prostate Cancer. Cancer Epidemiol Biomarkers Prev. 2003;12(12):1403–9.

- Abbaoui B, Lucas CR, Riedl KM, Clinton SK, Mortazavi A. Cruciferous Vegetables, Isothiocyanates, and Bladder Cancer Prevention. Mol Nutr Food Res. 2018;62(18):1–14. DOI: https://doi.org/10.1002/mnfr.201800079

- Zhao H, Lin J, Grossman HB, Hernandez LM, Dinney CP, Wu X. Dietary isothiocyanates, GSTM1, GSTT1, NAT2 polymorphisms and bladder cancer risk. Int J Cancer. 2007;120(10):2208–13. DOI: https://doi.org/10.1002/ijc.22549

- Mastuo T, Miyata Y, Yuno T, Mukae Y, Otsubo A, Mitsunari K, et al. Molecular mechanisms of the anti-cancer effects of isothiocyanates from cruciferous vegetables in bladder cancer. Molecules. 2020;25(3). DOI: https://doi.org/10.3390/molecules25030575

- Omoja V, Ihedioha T, Eke G, PeterAjuzie I, Okezie S. Evaluation of the acute toxicity, phytochemical constituents and anti - ulcer properties of methanolic leaf extract of Annona muricata in mice. J Intercult Ethnopharmacol. 2014;3(1):37. DOI: https://doi.org/10.5455/jice.20140111103203

- Evans W.C. Trease and Evan's Pharmacognosy. Haroourt Publishers,London, ISBN: 0702026174, 336-337.

- Harborne J.B. Phytochemicsal methods: A Guide to Modern Techniques of Plant Analysis. London, Chapman and Hall, 2007; ISBN: 81-8128-410-4, 1-34.

- Khalaf HA, Mahdi MF, Abaas IS. Preliminary Phytochemical and GC-MS analysis of chemical constituents of Iraqi Plantago lanceoleta L. Al-Mustansiriyah Journal of Pharmaceutical Sciences (AJPS) (2018); 18.2: 114-121. DOI: https://doi.org/10.32947/ajps.v18i2.485

- Souza BA. Manual de ensaios toxicológicos in vivo.Manual de ensaios toxicológicos in vivo. 1994. 122-122.‏‏

- Bioassay Technology Labrotary kit protocol available from : (www.bt-laboratory.com | 1008 Junjiang Inter. Bldg. 228 Ningguo Rd. Yangpu Dist. Shanghai. China Tel: 86 21 65109711 | Fax: 86 21 65109711 816 | E-mail: [email protected])

- Anthony L. Junqueira’s Basic Histology text and atlas. Edition. McGraw-Hill Education. 2016

- Gaskill, C. L., et al. "Liver histopathology and liver and serum alanine aminotransferase and alkaline phosphatase activities in epileptic dogs receiving phenobarbital." Veterinary Pathology 42.2 (2005): 147-160

- Balogh J, Iii DV, Gordon S, Li X, Ghobrial RM, Jr HPM. Jhc-3-041. J Hepatocell Carcinoma [Internet]. 2016;Volume 3:41–53. Available from: https://www.dovepress.com/hepatocellular-carcinoma-a-review-peer-reviewed-article-JHC. DOI: https://doi.org/10.2147/JHC.S61146

- Kim E, Viatour P. Hepatocellular carcinoma: old friends and new tricks. Exp Mol Med [Internet]. 2020;52(12):1898–907. Available from: http://dx.doi.org/10.1038/s12276-020-00527-1 DOI: https://doi.org/10.1038/s12276-020-00527-1

- Ahmed AG, Hussein UK, Ahmed AE, Kim KM, Mahmoud HM, Hammouda O, et al. Mustard Seed (Brassica nigra) Extract Exhibits Antiproliferative Effect against Human Lung Cancer Cells through Differential Regulation of Apoptosis, Cell Cycle, Migration, and Invasion. Molecules 2020;25(9):2069. Available from: http://dx.doi.org/10.3390/molecules25092069

- Verhoeven DTH, Goldbohm RA, Van Poppel G, Verhagen H, Van Den Brandt PA. Epidemiological studies on Brassica vegetables and cancer risk. Cancer Epidemiol Biomarkers Prev. 1996;5(9):733–48.

- Kristal AR. Brassica vegetables and prostate cancer risk: A review of the epidemiologic evidence. Pharm Biol. 2002;40(SUPPL.):55–8. DOI: https://doi.org/10.1076/phbi.40.7.55.9169

- Nandini T, Ramya Sree D, Janbee S, Siva Sai Kiran K, Raveendra Babu G. Study of immunomodulatory effect of seeds of Brassica nigra and Cuminum cyminum in albino rats. Int J Pharm Clin Res. 2016;8(1):10–4.

- Ahmed, Asmaa Gamal, et al. "Mustard seed (Brassica nigra) extract exhibits antiproliferative effect against human lung cancer cells through differential regulation of apoptosis, cell cycle, migration, and invasion." Molecules 25.9 (2020): 2069. DOI: https://doi.org/10.3390/molecules25092069

- Upwar N, Patel R , Waseem N , Mahobia N. In vitro anthelmintic activity of Brassica nigra Linn. seeds. (2011).

- Anand P, Murali YK, Tandon V, Murthy PS, Chandra R. Insulinotropic effect of aqueous extract of Brassica nigra improves glucose homeostasis in streptozotocin induced diabetic rats. Exp Clin Endocrinol Diabetes. 2009;117(6):251–6.‏ DOI: https://doi.org/10.1055/s-2008-1080917

- Basha SN, Rekha R, Saleh S, Yemane S. Evaluation of invitro anthelmintic activities of Brassica nigra, Ocimum basilicum and Rumex abyssinicus. Pharmacogn J. 2011;3(20):88–92. DOI: https://doi.org/10.5530/pj.2011.20.17

- Radha Krishnan K, Azhagu Saravana Babu P, Babuskin S, Sivarajan M, Sukumar M. Modeling the Kinetics of Antioxidant Extraction from Origanum vulgare and Brassica nigra. Chem Eng Commun. 2015;202(12):1577–85. DOI: https://doi.org/10.1080/00986445.2014.957757

- Uddin MS, Millat MS, Islam MS, Hussain MS, Uddin MG, Siddiqui SA, et al. Exploration of in vitro thrombolytic, anthelminthic, cytotoxic and in vivo anxiolytic potentials with phytochemical screening of flowers of Brassica nigra. Futur J Pharm Sci. 2020;6(1). DOI: https://doi.org/10.1186/s43094-020-00099-x

- Danlami U. Phytochemical, Nutritional and Antimicrobial Evaluations of the Aqueous Extract of Brassica Nigra (Brassicaceae) Seeds. 2016; 4(4):161. DOI: https://doi.org/10.11648/j.ajac.20160404.17

- Sharma A, Rai PK, Prasad S. GC–MS detection and determination of major volatile compounds in Brassica juncea L. leaves and seeds. Microchem J [Internet]. 2018;138(2017):488–93. Available from: https://doi.org/10.1016/j.microc.2018.01.015 DOI: https://doi.org/10.1016/j.microc.2018.01.015

- Organisation for Econommic Cooperation and Development,OECD Guidline for the testingof chemicals.Test guidline 423, acute oral toxicity –acute toxic class method.http://www.oecdilibrary.org.

- Arogundade TT, Enaibe BU, Adigun OO, Adigun FM, Gbadamosi IT. Dose-dependent effect of black mustard seeds (Brassica nigra) extract on the prefrontal cortex of adult Wistar rats. Anatomy. 2017;11(3):107–14. DOI: https://doi.org/10.2399/ana.17.027

- Kumar M, Sharma S, Vasudeva N. In vivo assessment of antihyperglycemic and antioxidant activity from oil of seeds of Brassica nigra in streptozotocin induced diabetic rats. Adv Pharm Bull. 2013;3(2):359–65.

- Jie M, Cheung WM, Yu V, Zhou Y, Tong PH, Ho JW. Anti-proliferative activities of sinigrin on carcinogen-induced hepatotoxicity in rats. PLoS One. 2014 Oct 20;9(10):e110145. doi: 10.1371/journal.pone.0110145. PMID: 25329483; PMCID: PMC4203766. DOI: https://doi.org/10.1371/journal.pone.0110145

- Elcombe CR, Odum J, Foster JR, Stone S, Hasmall S, Soames AR, et al. Prediction of rodent nongenotoxic carcinogenesis: Evaluation to biochemical and tissue changes in rodents following exposure to nine nongenotoxic NTP carcinogens. Environ Health Perspect. 2002;110(4):363–75. DOI: https://doi.org/10.1289/ehp.02110363

- Maronpot RR, Yoshizawa K, Nyska A, Harada T, Flake G, Mueller G, et al. Liver enlargement - STP regulatory policy papers: Hepatic enzyme induction: Histopathology. Toxicol Pathol. 2010;38(5):776–95. DOI: https://doi.org/10.1177/0192623310373778

- Wei, P., Zhang, J., Egan-Hafley, M. et al. The nuclear receptor CAR mediates specific xenobiotic induction of drug metabolism. Nature 2000; 407, 920–923 https://doi.org/10.1038/35038112 DOI: https://doi.org/10.1038/35038112

- Nesnow S, Ward W, Moore T, Ren H, Hester SD. Discrimination of tumorigenic triazole conazoles from phenobarbital by transcriptional analyses of mouse liver gene expression. Toxicol Sci. 2009;110(1):68–83. DOI: https://doi.org/10.1093/toxsci/kfp076

- Jones HB, Orton TC, Lake BG. Effect of chronic phenobarbitone administration on liver tumour formation in the C57BL/10J mouse. Food Chem Toxicol. 2009 Jun;47(6):1333-40. doi: 10.1016/j.fct.2009.03.014. Epub 2009 Mar 17. PMID: 19298838. DOI: https://doi.org/10.1016/j.fct.2009.03.014

- Waterman CL, Currie RA, Cottrell LA, Dow J, Wright J, Waterfield CJ, et al. An integrated functional genomic study of acute phenobarbital exposure in the rat. BMC Genomics. 2010;11(1):1–17. DOI: https://doi.org/10.1186/1471-2164-11-9

- Geter DR, Bhat VS, Bhaskar Gollapudi B, Sura R, Hester SD. Dose-response modeling of early molecular and cellular key events in the CAR-mediated hepatocarcinogenesis pathway. Toxicol Sci. 2014;138(2):425–45. DOI: https://doi.org/10.1093/toxsci/kfu014

- KILIMCI, Figen SEVIL. Rabbit liver lobes: an anatomical study of experimental surgical approaches. Erciyes Üniversitesi Veteriner Fakültesi Dergisi, 2020, 17.2: 103-108. DOI: https://doi.org/10.32707/ercivet.760679

- Health H, Meeting J, Panel FAO, Residues P, Core WHO, Group A, et al. ”Interpretation of liver effects” Provided by UK. 2018;1–4.

- Melega S, Canistro D, De Nicola GR, Lazzeri L, Sapone A, Paolini M. Protective effect of Tuscan black cabbage sprout extract against serum lipid increase and perturbations of liver antioxidant and detoxifying enzymes in rats fed a high-fat diet. Br J Nutr. 2013;110(6):988–97. DOI: https://doi.org/10.1017/S0007114513000068

- Gaskill CL, Miller LM, Mattoon JS, Hoffmann WE, Burton SA, Gelens HCJ, et al. Liver histopathology and liver and serum alanine aminotransferase and alkaline phosphatase activities in epileptic dogs receiving phenobarbital. Vet Pathol. 2005;42(2):147–60. DOI: https://doi.org/10.1354/vp.42-2-147

- Müller PB, Taboada J, Hosgood G, Partington BP, VanSteenhouse JL, Taylor HW, et al. Effects of long-term phenobarbital treatment on the liver in dogs. J Vet Intern Med. 2000;14(2):165–71. DOI: https://doi.org/10.1111/j.1939-1676.2000.tb02231.x

- Rajamurugan R, Selvaganabathy N, Kumaravel S, Ramamurthy C,Sujatha V, Thirunavukkarasu C. Polyphenol contents and antioxidant activity of Brassica nigra (L.) Koch. leaf extract. NatProd Res.(2011).

- Rajamurugan R, Selvaganabathy N, Kumaravel S, Ramamurthy C, Sujatha V, Thirunavukkarasu C. Polyphenol contents and antioxidant activity of Brassica nigra (L.) Koch. leaf extract. Nat Prod Res. 2012;26(23):2208–10 DOI: https://doi.org/10.1080/14786419.2011.637215

- Ahn M, Kim J, Bang H, Moon J, Kim GO, Shin T. Hepatoprotective effects of allyl isothiocyanate against carbon tetrachloride-induced hepatotoxicity in rat. Chem Biol Interact. 2016;254:102–8. Available from: http://dx.doi.org/10.1016/j.cbi.2016.05.037 DOI: https://doi.org/10.1016/j.cbi.2016.05.037

- Radwan SA, El-wessemy AM, Sakr SM. Sodium Barbital Induced Biochemical , Histological and Histochemical Changes in the Liver of Albino Mouse * Department of Biological Sciences , Faculty of Education , Ain Shams University , Cairo , Egypt . ** High Institution of Applied arts , 6 th October City Abstract. 2000;(1971).

- Mansour A, Abdel-Raheem K, El-Mossallamy N, El-Yamany N , Mahmoud S .Impact of chronic administration of ethanol and/or phenobarb-itone and drug withdrawal on transaminases and proteins in blood and liver of albino rats. J. Egypt. Ger. Soc. Zool.1995;16 (A): 429-450.

- Asaad NK, Razooqi QA, Mustafa MA. Toxicity of Cadmium Chloride on White Rats Liver and the Protective Role of Brassica Nigra Seed Extract. Indian J Forensic Med Toxicol. 2021;(January).