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Effect of Multiflora Honey on VEGF Expression and Renal Glomerular Tuft Area

This study examines the effect of multiflora honey on VEGF expression and glomerular tuft area in male Sprague-dawley rats induced with STZ. The aim is to determine if multiflora honey can decrease VEGF expression and glomerular tuft area.

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Effect of Multiflora Honey on VEGF Expression and Renal Glomerular Tuft Area

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  1. Traditional Medicine-2015Birmingham, UKAugust 03 – 05, 2015 Traditional Medicine 2015 SUSILORINI

  2. 3rd International Conference and Exhibition on  Traditional & Alternative Medicine August 03-05, 2015 Birmingham, UK In Association with: Presented By Name: SUSILORINI Country: INDONESIA

  3. EFFECT OF MULTIFLORA HONEY ON VEGF EXPRESSION AND RENAL GLOMERULAR TUFT AREA Experimental Study on Streptozotocin-induced Male Sprague-dawley Rats Susilorini* Indra Wijaya** Nur Wijaya H

  4. Overview • Background • Hypothesis • Methods • Results • Discussion • Conclusions & Future Directions

  5. Diabetic nephropathy is a diabetic complication that leads to renal disease and is one of thefactors of pain and death of diabetecian (10 years survival rate : 14% DM type 1: 33% DM type 2) • WHO predicts increasing number of diabetic nephropathy in the beginning of twenty first century. • In the developing countries there are 20-40% diabetic nephropathy in and 1% in child diabetecian in both types DM I and DM II.

  6. Many researchs has done to obtain effective drug, one of which is the honey • Some reseachers sugest that honey may lower blood glucose level in diabetician • Polifenol of multiflora honey especially quercetin is known to have antioxidant , anti-inflammation and anti apoptotic effects, which potentially reduce renal damage caused by diabetic nephropathy.

  7. Muliflora honey fromApis mellifera at Grinsing Batang, Central Java Flavonoid (0,21%) : total phenol (1,66%),quercetin (72,546%) glukose (350.593,34 ppm) , fruktose(465.634,35 ppm). Others: vit E (0,03 ppm), protein ( 0,09%) Cu ( 0,77 ppm), Mg (27,67ppm)..

  8. The aim of this study The research is carried out to determine the effect of multiflora honey on VEGF expression and glomerular tuft area on male Sprague-dawleyrats induced with STZ.

  9. Overview • Background • Hypothesis • Methods • Results • Discussion • Conclusions & Future Directions

  10. Hypothesis • Multiflora honey will decrease VEGF expression and Gomerular tuft area

  11. Overview • Background • Hypothesis • Methods • Results • Discussion • Conclusions & Future Directions

  12. METHOD • Twenty rats are divided into four group, all are induced with single dose STZ of 50 mg/kg. Hyperglycemic status is confirmed with GDP level after two days. • Group P1, P2 and P3 are given the honey in graded-dose during fourteen days. • Rats are terminated on day fifteen. VEGF expression is observed using Alfred Score.

  13. K O1 O3,4,,5 P1,2,3 Sampel Random Research design • Experimental, randomized post test only control group design

  14. VEGF Expression was analyzed using Kruskal Wallis test continued with Mann Whitney U Post-hoc test and Spearman correlation test. • Glomerular tuft area was measured microscopically. Glomerular tuft area was tested using One-way Anova Test continued with post hoc LSD and Pearson Correlation Test.

  15. Overview • Background • Hypothesis • Methods • Results • Discussion • Conclusions & Future Directions

  16. Mann Whitney U test result shows differences between controlled group and group P1 and P2, between group P1 and P3 and between group P2 ad P3. No difference between controlled group and group P3, and between group P1 and P2.

  17. LSD post hoc test result difference between controlled group and treated group P1, P2 and P3. But there is no difference within the group

  18. RESULT • VEGF expressions on group P1 and P2 are significantly different from control group, but group P3 (p= 0,0001) is not. • Significant differences of glomerular tuft area occur between control group and the other groups P1, P2 and P3 (p= 0,0001). • There are no differences among groups P1, P2 and P3. There is a dose effect relationship of glomerular tuft area with r= -0, 291. • Graded dose multiflora honey (0,33 ; 1 and 10 g/kgBW/day) significantly given lower glomerular tuft area , but lower VEGF expression was only seen in the group of 0 ,33 and 1 g/kgBW/day.

  19. Hasil 9 Picture 1. Histopathological morfologi of the kidney (H.E staining) K P1 normal P2 P3

  20. Picture 2. VEGF expression . (IHC usinganti-VEGF-A staining) K P1 P2 P3 normal

  21. Overview • Background • Hypothesis • Methods • Results • Discussion • Conclusions & Future Directions

  22. Multiflora honey is honey produced by bees consuming nectar from various flowers. • It has the highest polyphenol content so is expected to have the highest antioxidant effect. • Some researches (Erejewa et al, Khalil, et al; Al Wali) clinically proved that honey has low glycemic index and is safe on both DM type I and II

  23. Overview • Background • Hypothesis • Methods • Results • Discussion • Conclusions & Future Directions

  24. Hyperglicemia Metabolic change: OKSIDATIVE STRESS Hemodinamic change Structural change of the kidney: podositopati Increased VEGF expression hypertrophi of the glomerulus ( area glomerular tuft)

  25. quercetin Vitamin E

  26. Honey contains vitamin A, C, E, organic acid, enzymes, minerals, phenol and flavonoid as antioxidant, anti-apoptosis and anti-inflammation and also free radicals scavenger (Astarika, 2011). • Honey supplementary in a correct way decrease MDA level and correct SOD and CAT activity in pancreas of diabetic rats ( Erejuwa & friends, 2012).

  27. Quercetin prevents oxidative damage and cell death through some mechanism including oxidative stress scavenger, chelating metalic ions (Chen et al, 1990), antiinflammation by inhibited xanthine oxidase (Chang et al, 1993) and lipid peroxidase (Molan et al, 2000), and anti-apoptosis (saija et al, 1995; Miller, 1996; Cox et all, 2000),

  28. CONCLUSION Multiflora honey ( 0,33 ml/kgBW/day and 1 ml/kgBW/day) are proven to be able decreasing VEGF expression. • Multiflora honey ; • All dosage (0,33 ml/kgBW/day, 1 ml/kgBW/day and 10 ml/kgBW/day) is able to lower glomerular tuft area.

  29. Further and deeper research with more sample is needed so the result can be applied clinically and follow up research is also needed with longer timeto study chronic renal morphology changes.

  30. References • Sudoyo AW, Setiyohadi B, Alwi I, Simadibrata M, Setiati S. Buku Ajar IlmuPenyakitDalam. Editor: DepartemenIlmuPenyakitDalamFakultasKedokteran Indonesia. .Jakarta: Penerbit UI, 2006; Edisi IV, jilid III. Hal:1892-1907 • World Health Organization. Definition and diagnosis of diabetes mellitus and intermediate hyperglycemia: report of a WHO/IDF consultation. Geneva: WHO Press, World Health Organization.. 2006;20. • Wolf G, Ziyadeh F, Cellular and molecular mechanisms of proteinuria in diabetic nephropathy. Nephron Physiol. 2007;106:26-31. • Sustztak K, Raff A, Schiffer M, Botinger E. Glucosed- induced Reactive Oxygen Species Cause Apoptosis of Podocytes and Podocyte Depletion at the Onset of Diabetic Nephropathy. Diab J. 2006; 55:225-33 • Sourris KC and Forbes JM.Pathology RAGE Drives the Development of Glomerulosclerosis and Implicates Podocyte Activation in the Pathogenesis of Diabetic Nephropathy Interactions Between Advanced Glycation End-Products (AGE) and their Receptors in the Development and Progression of Diabetic Nephropathy. Am J Path. .2008; 162( 4): 1110-17 • Al-Waili NS. Natural honey lowers plasma glucosetikan, homocysteine, and blood lipids in healthy, diabetic, and hyperlipidemic subjects: comparison with dextrose and sucrose. J of Med Food. 2004; 7(1):100-7. • Chepulis LM. An Investigation of the Health Benefits of Honey as a Replacement For Sugar In the Diet. J of Comp Int Med. 2005; 4(1): 8. • Menini, et al. Increased glomerular cell (podocyte) apoptosis in rats with streptozotocin-induced diabetes mellitus: role in the development of diabetic glomerular diseases. Diabetologia. 2007 September 50:2591–9. • Ziyadeh FN, Wolf G. Pathogenesis of the podocytopathy and proteinuria in diabetic glomerulopathy. Curr Diabetes Rev. 2008.;Feb;4(1):39-45. • Consentino F, Eto M, Paola de Paolis, Ben van derLoo, Bachscimd M, Ulrich V, Kouroedov, et al. High Glucose Causes Upregulation of Cyclooxygenase-2 and Alters Prostanoid Profile in Human Endothelial Cells: Role of Protein Kinase C and Reactive Oxygen Species.Circulation. 2003; 107: 10017-23. (cited on february 27, 2010). http://www.circ.ahajournals.org • Sho-ichi Y, Matsui T. Advanced glycation end products, oxidative stress and diabetic nephropathy. Oxid Med Cell Longev.2010. Mar-Apr;3 (2); 101-8. • Brosius FC, Khoury CC, Buller CL, Chen S. Abnormalities in signaling pathways in diabetic nephropathy. Expert Rev EndocrinolMetab. 2010; 5(1): 51-64. • Guan F, Villegas G, Teichman J, Mundel T, Tufro A. Autocrine VEGF-A system in podocytes regulates podocin and its interaction with CD2AP. Am J Physiol Renal Physiol. 2006. 291:422-28. • Khalil M, Sulaiman S, Boukra L. Antioxidant properties of honey and its role in preventing health disorder. The Open Neu J. 2010; 3:6-16. • Gheldof N, Engeseth NJ. Antioxidant capacity of honeys from various floral sources based on the determination of oxygen radical absorbance capacity and inhibition of in vitro lipoprotein oxidation in human serum samples. J Agric Food Res. 2002; 50: 3050-5. • Jaganathan SK, Mandal M. A review: Antiproliferatif effects of honey and its polyphenol. J. Biomed and Biotech. 2009;1-3. • Busserroles, Geux, Rock, Mazur and Rayssiguler. Substituting honey for refined carbohydrates protects rats from hypertriglyseridemic and prooxidative effects of fructose.J Am Soc Sci.2002;3379-81 • Erejuwa O, et al. Antioxidant protective effect of glibenclamide and metformin in combination with honey in kidney of streptozotocin- induced diabetic rats. Int J Mol Sci. 2011;12,829-43. • Schrijvers BF, Flyvbjerg A, Vriese A. the role of vascular endothelial growth factor ( VEGF) in renal pathophysiology. Kidney Int J..2004;65:2003-17.(cited on 23 february 2010). Available from URL: http://www.ncbi.nlm.nih.gov/guide/PubMedresult..

  31. THANK YOU t

  32. Program Pascasarjana Magister Ilmu Biomedik Dan Pendidikan Dokter Spesialis I Ilmu Patologi Anatomi Fakultas Kedokteran Universitas Diponegoro Semarang

  33. Meet the eminent gathering once again atTraditional Medicine-2016London, UKOctober 03-05, 2016 Traditional Medicine-2016 Website: http://traditionalmedicine.conferenceseries.com/

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