STZ (Streptozotosin) ile Diyabet Oluşturulmuş Ratların Böbrek Dokusunda Silibinin'in HIF-1? (Hypoxia-InducibleFactor-1 Alpha) ve TLR-2 (Toll Like Receptor 2) Genlerinin mRNA Düzeylerine Etkisi

Silibinin, Silybum marianum (Deve dikeni) adı verilen bitkiden elde edilen bir flavanoiddir. Bu deneysel çalışmada; hücre koruyucu, antiinflamatuar, antikanserojen, antioksidan etkileri olan ve proteüniriyi azalttığı bilinen Silibininin, STZ ile diyabet oluşturulan ratların böbreklerinde HIF-1? ve TLR2 genlerinin ekspresyon seviyelerine etkisini tedavi öncesi ve sonrası karşılaştırıp, histokimyasal olarak da inceledik. Çalışmamızda 5 grup vardı: Kontrol Grubu, Diyabet Grubu (STZ 65 mg/kg, i.p. tek doz), Diyabetik Grup+100mg/kg Silibinin Tedavi Grubu, Diyabetik Grup+200mg/kg Silibinin Tedavi Grubu, Silibinin grubu (5 rata 100 mg/kg silibinin, 5 rata 200 mg/kg silibinin). Böbreğin biri HIF-1? ve TLR2 genlerinin ekspresyon seviyeleri için değerlendirilirken, diğer böbrek histopatolojik olarak değerlendirildi. Histolojik olarak ilaç alan gruplarda, diyabetik gruplara kıyasla Diyabetik Nefropati (DN) bulguları açısından anlamlı bir değişiklik gözlenmedi. Alınan doku örneklerinden elde edilen total RNA'lar kullanılarak RT-PCR metodu ile 2 adet hedef gen ve bir adet kalibratör genin ifadesindeki değişiklikler incelendi. Elde edilen bulgular kontrol grubu ve diğer gruplardaki örnekler ile karşılaştırılmıştır. 2-??CT yöntemi ile kat değişimleri hesaplandığında ve eşik -2/2 olarak alındığında TLR2 geninde değişimler tespit edilmiştir ancak istatistiksel olarak anlamlı bir fark bulunamamıştır (p< 0,05). Silibinin'in diyabetli ratların böbrek dokusunda oluşan hasara karşı yeteri kadar etkili olmadığı görüldü.

Effect of Silibinin on mRNA Levels of HIF-1? (Hypoxia-Inducible Factor-1 Alpha) and TLR-2 (Toll-Like Receptor 2) Genes in Kidney of STZ (Streptozotocin) Induced Diabetic Rats

Silibinin is a flavonoid that derived from the plant named Silybum marianum.In this experimental study, we compared and histochemically examined the effect of Silibinin on the expression levels of HIF-1? and TLR2 genes in the kidneys of STZ-diabetic rats before and after the treatment. There were 5 groups in this study: Control Group, Diabetes Group (STZ 65 mg/kg, i.p. single dose), Diabetic Group+100mg/kg Silibinin Treatment Group, Diabetic Group +200mg/kg Silibinin Treatment Group, Silibinin group (100 mg/kg silibinin on 5 rats, 200 mg/kg silibinin on 5 rats). The changes in the expression of two target genes and one calibrator gene were examined using the RT-PCR method. No statistically significant difference was found when the fold changes were calculated using 2-??CT method (p>0,05). No significant change was observed in the groups taking drugs histologically when compared to diabetic groups. It was seen that Silibinin is not quite effective on the damage formed in the kidney tissue of diabetic rats according to Diabetic Nephropaty findings. Also silibinin is not effective on the mRNA levels of diabetic rat kidneys.

PDF

___

1. Jorgens, V. (2006). Oskar Minkowski (1858-1931). An outstanding master of diabetes research. HORMONES-ATHENS-, 5(4), 310.
2. Zimmet, P. Z., Magliano, D. J., Herman, W. H., & Shaw, J. E. (2014). Diabetes: a 21st century challenge. The lancet Diabetes & endocrinology, 2(1), 56-64.
3. Lu, J., Jaafer, R., Bonnavion, R., Bertolino, P., & Zhang, C. X. (2014). Transdifferentiation of pancreatic ?-cells into insulin-secreting cells: From experimental models to underlying mechanisms. World journal of diabetes,5(6), 847.
4. Lin, H. P., Chan, T. M., Fu, R. H., Chuu, C. P., Chiu, S. C., Tseng, Y. H., ... & Chen, H. S. (2015). Applicability of adipose-derived stem cells in Type 1 diabetes mellitus. Cell transplantation, 24(3), 521-532.
5. Forouhi, N. G., & Wareham, N. J. (2014). Epidemiology of diabetes. Medicine,42(12), 698- 702.
6. Chowdhury, T. A., Shaho, S., & Moolla, A. (2014). Complications of diabetes: progress, but significant challenges ahead. Annals of translational medicine,2(12).
7. Hu, C., Sun, L., Xiao, L., Han, Y., Fu, X., Xiong, X., ... & Kanwar, Y. S. (2015). Insights into the mechanisms involved in the expression and regulation of extracellular matrix proteins in diabetic nephropathy. Current medicinal chemistry, 22(24), 2858-2870.
8. Martínez-Castelao, A., Navarro-González, J. F., Górriz, J. L., & de Alvaro, F. (2015). The concept and the epidemiology of diabetic nephropathy have changed in recent years. Journal of clinical medicine, 4(6), 1207-1216.
9. Ullah, F., Afridi, A. K., Rahim, F., Ashfaq, M., Khan, S., Shabbier, G., & ur Rahman, S. (2015). Knowledge of diabetic complications in patients with diabetes mellitus. Journal of Ayub Medical College Abbottabad, 27(2), 360-363.
10. López-Revuelta, K., Abreu, A. A. M., GerreroMárquez, C., Stanescu, R. I., Marín, M. I. M., & Fernández, E. P. (2015). Diabetic Nephropathy without Diabetes. Journal of clinical medicine, 4(7), 1403-1427.
11. Dasu, M. R., Devaraj, S., Park, S., & Jialal, I. (2010). Increased toll-like receptor (TLR) activation and TLR ligands in recently diagnosed type 2 diabetic subjects. Diabetes care, 33(4), 861- 868.
12. Ziello, J. E., Jovin, I. S., & Huang, Y. (2007). Hypoxia-Inducible Factor (HIF)-1 regulatory pathway and its potential for therapeutic intervention in malignancy and ischemia. Yale J Biol Med, 80(2), 51-60.
13. Takiyama, Y., & Haneda, M. (2014). Hypoxia in diabetic kidneys. BioMed research international, 2014.
14. Kren, V., & Walterova, D. (2005). Silybin and silymarin-new effects and applications. Biomedical Papers, 149(1), 29-41.
15. Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2- ??CT method. methods,25(4), 402-408.
16. Xu, X., Chen, P., Zheng, Q., Wang, Y., & Chen, W. (2011). Effect of pioglitazone on diabetic nephropathy and expression of HIF-1? and VEGF in the renal tissues of type 2 diabetic rats. Diabetes research and clinical practice, 93(1), 63-69.
17. Chen, S., Okahara, F., Osaki, N., & Shimotoyodome, A. (2015). Increased GIP signaling induces adipose inflammation via a HIF1?-dependent pathway and impairs insulin sensitivity in mice. American Journal of Physiology-Endocrinology and Metabolism, 308(5), E414-E425.
18. Girgis, C. M., Cheng, K., Scott, C. H., & Gunton, J. E. (2012). Novel links between HIFs, type 2 diabetes, and metabolic syndrome. Trends in Endocrinology & Metabolism, 23(8), 372-380.
19. Tang, S. C., Yiu, W. H., Lin, M., & Lai, K. N. (2015). Diabetic nephropathy and proximal tubular damage. Journal of Renal Nutrition, 25(2), 230- 233.
20. Rajamani, U., & Jialal, I. (2014). Hyperglycemia induces Toll-like receptor-2 and-4 expression and activity in human microvascular retinal endothelial cells: implications for diabetic retinopathy. Journal of diabetes research, 2014.
21. Sawa, Y., Takata, S., Hatakeyama, Y., Ishikawa, H., & Tsuruga, E. (2014). Expression of toll-like receptor 2 in glomerular endothelial cells and promotion of diabetic nephropathy by Porphyromonas gingivalis lipopolysaccharide. PloS one, 9(5), e97165.
22. Zikou, X., Tellis, C. C., Rousouli, K., Dounousi, E., Siamopoulos, K. C., & Tselepis, A. D. (2015). Differential membrane expression of toll-like receptors and intracellular cytokine induction in peripheral blood monocytes of patients with chronic kidney disease and diabetic nephropathy. Nephron Clinical Practice, 128(3-4), 399-406.