Evaluation of the effect of honey on immune status in mice bearing Ehrlich carcinoma

1. بسم الله الرحمن الرحيم Evaluation of the effect of honey on immune status in mice bearing Ehrlich carcinoma Ahmed G. Hegazi; Eman H. Abdel- Rahman; Samira Shawky, FayrouzAbd Allah and Amr M. Abdou National Research Center, Dokki, Giza, Egypt ahmed@ahmedhegazi com and ahmedhegazi128@gmail.com www.ahmedhegazi.com

4. Introduction • Cancer continues to represent the largest cause of mortality in the world and claims over 6 million lives every year (Abdullaev et al., 2000). • An extremely promising strategy for cancer prevention today is chemoprevention.

5. Plants, vegetables and herbs used in the folk and traditional medicine have been accepted currently as one of the main sources of cancer chemoprevention drug discovery and development (Abdullaev, 2001).

6. Antioxidants play an important role in inhibiting and scavenging radicals, thus providing protection to humans against infection and degenerative diseases (Hegazi & Abd El Hady, 2009 and Hegazi et al., 2014).

7. There is a growing interest in the pharmacological evaluation of various natural products used in traditional medicine. • Flavonoids, terpenoids, and steroids have received considerable attention in recent years due to their diverse pharmacological properties including antioxidant and antitumor activity (DeFeudis et al., 2003 and Takeoka & Dao, 2003).

8. Aim

9. From this viewpoint the present study was carried out to • evaluate the antitumor activity, • immunoglobulin levels, • phagocytic activity • lymphocyte transformations • delayed hypersensitivity • of coriander honey against Ehrlich ascites carcinoma (EAC) in Swiss albino mice.

10. Material and Methods:

11. Coriander honey: • Coriander honey was purchased from local market in Egypt. • Sterile distilled water was used to dilute honey immediately before administration by a stomach tube. • Honey was used in a final concentration of 500 mg/kg/mouse.

12. Animals: Studies were carried out using a total of 210 male Swiss albino mice weighting 22- 25 g obtained from Animal House of National Research Center, Giza, Egypt.

13. Ehrlich ascites carcinoma: Ehrlich ascites carcinoma (EAC) cells were obtained from Cancer Biology Section, National Cancer Institute, Cairo, Egypt. The Ehrlich tumor line was maintained, till the time of the experiment in female Swiss albino mice by serial intraperitoneal passage of 2×106 cells/mouse at 7-10 days intervals (Hegazi et al., 2014).

14. Standard anticancer drug: 5-Fluorouracil (Calbiochem, USA) was used as a standard anticancer drug. 5-Fluorouracil was injected intraperitoneally to mice at a dose of 20 mg/kg body weight (Karthigayan et al., 2007 and Balamurugan et al., 2010).

15. Experimental design: • A total of 210 male Swiss albino mice were divided into 6 groups (n=35) with an average weight 22-25 gm each as follows: 

17. At the second day post inoculation with Ehrlich carcinoma 0.1 ml of bovine serum albumin was inoculated intraperitoneally in each mice in all groups as priming for the specific antigen used in the skin test. • Blood and serum samples were collected from 5 mice from each group at 7, 14 and 21 days intervals post inoculation with Ehrlich effusive carcinoma cells. • Blood samples were subjected to determine lymphocyte transformation as Hegazi et al. (1981) & (1985) • phagocytic activity as Hegazi et al. (1995). • While the sera sample were used for the detection of different Immunoglobulin levels IgG, IgA and IgM (Deverill, 1978).

18. After 14 days of the experiment 5 mice from each group were subjected to 18 h fasting before sacrificing. • Anti- tumor effect of coriander honey was assessed by • observation of changes in body weight, • ascetics volume, • viable and nonviable tumor cell count.

19. The remaining animals in each group were kept • to check the mean survival time (MST) of the tumor bearing hosts • and increase in life span percentage (ILS %). • MST of each group containing five mice were monitored by recording the mortality daily for 7 weeks and ILS % was calculated using the following equation: MST= (Day of first death + Day of last death)/2. ILS % = [(Mean survival time of treated group/mean survival time of control group) -1] X 100 (Mazumder et al., 1997 and Gupta et al., 2000).

20. Blood and serum samples were collected from 5 mice from each group at 7, 14 and 21 days intervals post inoculation with Ehrlich carcinoma cells. • Blood samples were subjected to determine • blood lymphocytic count, • lymphocyte transformation • The phagocytic activity • While the sera sample were used for detection of different Immunoglobulin levels by quantitative determination of IgG, IgA and IgM (Deverill, 1978).

21. At 72 hours before the end of the experiment a 0.1-ml of bovine serum albumin was inoculated in the left footpad of the mice where the right foot pad was inoculated with saline. • Skin thickness was measured, 24 hours and 72 hours post inoculation to evaluate the delayed type hypersensitivity as Cheville and Beard (1972); Hegazi (1981) and Hegazi et al. (1985).

22. Results

23. Influence of honey in mice bearing Ehrlich carcinoma on immune status

24. Influence of honey in mice bearing Ehrlich carcinoma on immune status

25. Influence of honey in mice bearing Ehrlich carcinoma on immune status

26. Influence of honey in mice bearing Ehrlich carcinoma on immune status

27. Influence of honey in mice bearing Ehrlich carcinoma on immune status

28. Influence of honey in mice bearing Ehrlich carcinoma on immune status Influence of honey in mice bearing Ehrlich carcinoma on immune status

35. Effect of coriander honey on hematological parameters in EAC bearing mice.

36. Effect of coriander honey on IgM level in mice- bearing Ehrlich carcinoma: 

37. Effect of coriander honey on IgG level in mice- bearing Ehrlich carcinoma: 

38. Effect of coriander honey on IgA level in mice- bearing Ehrlich carcinoma: 

39. Effect of coriander honey on phagocytic activity level in mice- bearing Ehrlich carcinoma: 

40. Effect of coriander honey on Lymphocyte transformation (stimulation index using PHA) level in mice- bearing Ehrlich carcinoma: 

41. Effect of coriander honey on Delayed hypersensitivity (skin index / mm thickness): in mice- bearing Ehrlich carcinoma: 

42. ”سَنُرِيهِمْ آيَاتِنَا فِي الاَفَاقِ وَفِيَ أَنفُسِهِمْ حَتّىَ يَتَبَيّنَ لَهُمْ أَنّهُ الْحَقّأَوَلَمْ يَكْفِ بِرَبّكَ أَنّهُ عَلَىَ كُلّ شَيْءٍ شَهِيدٌ“ فصلت 53.

44. Evaluation of the effect of honey on immune status in mice bearing Ehrlich carcinoma

45. Evaluation of the effect of honey on immune status in mice bearing Ehrlich carcinoma Thank you

46. Prof. Dr. Ahmed Hegazi • Professor of Microbiology and ImmunologyNational Research Center, Dokki, Giza, Egypt • President of Egyptian Environmental Society for uses and production of bee productsSecretary of Egyptian Society of ApitherapySecretary General of African  Federation of Apiculture AssociationsMember of Apitherapy Commission , APIMONDIAE mail: ahmed@ahmedhegazi.com and ahmedhegazi128@gmail.com www.ahmedhegazi.com Tel:     +  2023 7749222 and + 20238860682 Fax:   + 202 37749222 and + 202 33370931 GMS: + 201001440063