Anat Cell Biol.  2019 Jun;52(2):183-190. 10.5115/acb.2019.52.2.183.

Genistein improve nicotine toxicity on male mice pancreas

• Salahshoor MR
• Mirzaei F
• Roshankhah S
• Jalili P
• Jalili C

• KMID: 2451222
• DOI: http://doi.org/10.5115/acb.2019.52.2.183

Abstract

Nicotine is the most toxic factor of tobacco. Genistein is a phytoestrogen and antioxidant that has numerous health benefits. The aim of this study is to evaluate the effects of genistein against toxic properties of nicotine to the pancreas of mice. For this purpose, 48 male mice were randomly assigned into six groups (n=8): normal control, nicotine control (2.5 mg/kg), genistein (25 and 50 mg/kg), and nicotine+genistein (25 and 50 mg/kg) treated groups. Various doses of genistein and genistein+nicotine were administered intraperitoneally to animals for 4 weeks. The weight of pancreas, total antioxidant capacity and nitrite oxide of serum, insulin levels, and the number and diameter of islets of Langerhans were investigated. Nicotine administration reduced significantly total antioxidant capacity, insulin, pancreas weight, and the number and diameter of islets of Langerhans and increased nitrite oxide in serum compared to the control normal group (P<0.05). Conversely, genistein and genistein+nicotine increased significantly insulin, total antioxidant capacity, and the number and diameter islets of Langerhans and decreased serum nitrite oxide compared to the nicotine control group. It seems that the genistein can improve pancreas damage following the nicotine administration.

Keyword

Genistein; Pancreas; Nicotine

MeSH Terms

Animals
Genistein*
Humans
Insulin
Insurance Benefits
Islets of Langerhans
Male*
Mice*
Nicotine*
Pancreas*
Phytoestrogens
Tobacco
Genistein
Insulin
Nicotine
Phytoestrogens

Figure

• 

  Fig. 1 Procedures for the dissection and remove of pancreas from the abdomen of one of the mice in treatment groups. (A) Stimulus examination. (B) Beginning removal of pancreas. (C) Pancreas extraction along the intestines. (D) Pancreas removal at the caecum. (E) Pancreas removal. (F) Pancreas examination, the pancreas with attached spleen being examined after removal from the mouse. Spleen is indicated by the red arrow, and the pancreas is indicated by the black arrow.

• 

  Fig. 2 Comparison of the mean of animal's weight between treatment groups. a)Significant decrease in the mean weight of pancreas in the nicotine control group (2.5 mg/kg) compared to the control normal (saline) group (P<0.05). b)Significant increase in both genistein groups compared to the nicotine control group (P<0.05). c)Significant increase compared to the nicotine control group and decrease compared to the both genistein and normal control groups (P<0.05). The mean weight of pancreas was not significant in all genistein groups compared to the normal control group (P>0.05).

• 

  Fig. 3 Effects of nicotine, genistein, and nicotine+genistein on the mean nitrite oxide levels. a)Significant increase in nitrite oxide level in the nicotine control group (2.5 mg/kg) compared to the normal control (saline) group (P<0.05). b)Significant decrease in both genistein groups compared to the nicotine control group (P<0.05). c)Significant decrease compared to nicotine control group and increased compared to the both genistein and normal control groups (P<0.05). The mean nitrite oxide level was not significant in all genistein groups compared to the normal control group (P>0.05).

• 

  Fig. 4 Correlation analysis between treatment groups in mean insulin level in blood serum. a)Significant decrease in insulin level in the nicotine control group compared to the normal control (saline) group (P<0.05). b)Significant increase in the mean of insulin level in blood serum n all genistein groups compared to the nicotine control group (P<0.05). c)Significant increase in insulin level in serum in both genistein+nicotine groups compared to the nicotine control group and decrease compared to the both genistein and normal control groups (P<0.05). The insulin in blood serum was not significant in all genistein groups compared to the normal control group (P>0.05).

• 

  Fig. 5 Correlation analysis between treatment groups in BALB/c male mice; morphometrically changes in pancreas islets number (A) and diameter of Langerhans islets (B). a)Significant decrease in the number and diameter of Langerhans islets in the nicotine control group (2.5 mg/kg) compared to the normal control group (saline) (P<0.05). b)Significant increase in the number and diameter of Langerhans islets in all genistein groups compared to the nicotine control group (P<0.05). c)Significant increase in of the number and diameter of Langerhans islets in all genistein+nicotine groups compared to the nicotine control group and decrease compared to the both genistein and normal control groups (P<0.05). The mean diameter and a number of pancreatic islets were as not significant in all genistein groups compared to the normal control group (P>0.05).

• 

  Fig. 6 Histological sections of the pancreas (H&E staining, ×100). (A) Normal pancreas structure in the micrograph of pancreas section in the normal control group. (B) Normal pancreas structure in the micrograph of pancreas section in genistein group (50 mg/kg). (C) after treatment with nicotine (2.5 mg/kg) in the nicotine control group, the pancreas section shows variable changes and manifest injury. Vacuolization in tissues (blue arrow), reduction of islet (red arrow), and blooding in pancreas tissue (yellow arrow) were observed. (D) Normal pancreas structure in the micrograph of pancreas section in nicotine+genistein group (50 mg/kg). After treatment with genistein in all doses, it was observed that genistein reduced pancreas damage caused by nicotine.

• 

  Fig. 7 Comparison of nicotine, saline, and genistein+nicotine groups with respect to the mean of serum total antioxidant capacity levels. a)Significant decrease in the mean of serum total antioxidant capacity level in the nicotine control group (2.5 mg/kg) compared to the normal control (saline) group (P<0.05). b)Significant increase in the mean of serum totals antioxidant capacity level at both genistein groups compared to the nicotine control group (P<0.05). c)Significant increase compared to the nicotine control group and decrease compared to the normal control group on the mean of serum total antioxidant capacity level in all genistein+nicotine groups (P<0.05).

Reference

1. Salahshoor MR, Roshankhah S, Hosseni P, Jalili C. Genistein improves liver damage in male mice exposed to morphine. Chin Med J (Engl). 2018; 131:1598–1604.

2. Velasquez MT, Bhathena SJ. Role of dietary soy protein in obesity. Int J Med Sci. 2007; 4:72–82.

3. Dolinoy DC, Weidman JR, Waterland RA, Jirtle RL. Maternal genistein alters coat color and protects Avy mouse offspring from obesity by modifying the fetal epigenome. Environ Health Perspect. 2006; 114:567–572.

4. Pavese JM, Farmer RL, Bergan RC. Inhibition of cancer cell invasion and metastasis by genistein. Cancer Metastasis Rev. 2010; 29:465–482.

5. Lee JS. Effects of soy protein and genistein on blood glucose, antioxidant enzyme activities, and lipid profile in streptozotocin-induced diabetic rats. Life Sci. 2006; 79:1578–1584.

6. Behloul N, Wu G. Genistein: a promising therapeutic agent for obesity and diabetes treatment. Eur J Pharmacol. 2013; 698:31–38.

7. Gawish AM, Ramadan S, Hassan AM, Issa AM. Morphometrical, histopathological, and cytogenetical ameliorating effects of green tea extract on nicotine toxicity of the testis of rats. J Cytol Histol. 2010; 1:1105.

8. Panahi S, Abdollahifar MA, Aliaghaei A, Nazarian H, Paktinat S, Abdi S, Farahani RM. Application of stereological methods for unbiased estimation of sperm morphology in the mice induced by busulfan. Anat Cell Biol. 2017; 50:301–305.

9. Kadiyala V, Lee LS, Banks PA, Suleiman S, Paulo JA, Wang W, Rosenblum J, Sainani NI, Mortele K, Conwell DL. Cigarette smoking impairs pancreatic duct cell bicarbonate secretion. JOP. 2013; 14:31–38.

10. Wang H, Yu M, Ochani M, Amella CA, Tanovic M, Susarla S, Li JH, Wang H, Yang H, Ulloa L, Al-Abed Y, Czura CJ, Tracey KJ. Nicotinic acetylcholine receptor alpha7 subunit is an essential regulator of inflammation. Nature. 2003; 421:384–388.

11. Polkowski K, Mazurek AP. Biological properties of genistein. A review of in vitro and in vivo data. Acta Pol Pharm. 2000; 57:135–155.

12. Jalili C, Salahshoor MR, Hoseini M, Roshankhah S, Sohrabi M, Shabanizadeh A. Protective effect of thymoquinone against morphine injuries to kidneys of mice. Iran J Kidney Dis. 2017; 11:142–150.

13. Wan C, Jin F, Du Y, Yang K, Yao L, Mei Z, Huang W. Genistein improves schistosomiasis liver granuloma and fibrosis via dampening NF-kB signaling in mice. Parasitol Res. 2017; 116:1165–1174.

14. Neuman JC, Truchan NA, Joseph JW, Kimple ME. A method for mouse pancreatic islet isolation and intracellular cAMP determination. J Vis Exp. 2014; e50374.

15. Elayat AA, el-Naggar MM, Tahir M. An immunocytochemical and morphometric study of the rat pancreatic islets. J Anat. 1995; 186(Pt 3):629–637.

16. Borai A, Livingstone C, Kaddam I, Ferns G. Selection of the appropriate method for the assessment of insulin resistance. BMC Med Res Methodol. 2011; 11:158.

17. Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem. 1996; 239:70–76.

18. Thrower E. Pathologic cellular events in smoking-related pancreatitis. Cancers (Basel). 2015; 7:723–735.

19. Huang H, Xu Y, van den Pol AN. Nicotine excites hypothalamic arcuate anorexigenic proopiomelanocortin neurons and orexigenic neuropeptide Y neurons: similarities and differences. J Neurophysiol. 2011; 106:1191–1202.

20. Mineur YS, Abizaid A, Rao Y, Salas R, DiLeone RJ, Gundisch D, Diano S, De Biasi M, Horvath TL, Gao XB, Picciotto MR. Nicotine decreases food intake through activation of POMC neurons. Science. 2011; 332:1330–1332.

21. Williams DL, Schwartz MW. The melanocortin system as a central integrator of direct and indirect controls of food intake. Am J Physiol Regul Integr Comp Physiol. 2005; 289:R2–R3.

22. Tamm C, Zhivotovsky B, Ceccatelli S. Caspase-2 activation in neural stem cells undergoing oxidative stress-induced apoptosis. Apoptosis. 2008; 13:354–363.

23. Hashemi J, Pasalar P, Soleimani M, Khorramirouz R, Fendereski K, Enderami SE, Kajbafzadeh AM. Application of a novel bioreactor for in vivo engineering of pancreas tissue. J Cell Physiol. 2018; 233:3805–3816.

24. Scuro LS, Simioni PU, Grabriel DL, Saviani EE, Modolo LV, Tamashiro WM, Salgado I. Suppression of nitric oxide production in mouse macrophages by soybean flavonoids accumulated in response to nitroprusside and fungal elicitation. BMC Biochem. 2004; 5:5.

25. Guo TL, Germolec DR, Zheng JF, Kooistra L, Auttachoat W, Smith MJ, White KL, Elmore SA. Genistein protects female nonobese diabetic mice from developing type 1 diabetes when fed a soy- and alfalfa-free diet. Toxicol Pathol. 2015; 43:435–448.

26. Jalili C, Ahmadi S, Roshankhah S, Salahshoor M. Effect of Genistein on reproductive parameter and serum nitric oxide levels in morphine-treated mice. Int J Reprod Biomed (Yazd). 2016; 14:95–102.

27. Gumustekin K, Ciftci M, Coban A, Altikat S, Aktas O, Gul M, Timur H, Dane S. Effects of nicotine and vitamin E on glucose 6-phosphate dehydrogenase activity in some rat tissues in vivo and in vitro. J Enzyme Inhib Med Chem. 2005; 20:497–502.

28. Fu Z, Zhang W, Zhen W, Lum H, Nadler J, Bassaganya-Riera J, Jia Z, Wang Y, Misra H, Liu D. Genistein induces pancreatic beta-cell proliferation through activation of multiple signaling pathways and prevents insulin-deficient diabetes in mice. Endocrinology. 2010; 151:3026–3037.