The Effect of Doxycycline on Glomerulosclerosis in 5/6 Renal Ablation Nephropathy

Abstract

Objectives: The beneficial effect of matrix metalloproteinases (MMP) inhibitors in renal diseases has been reported. Their effect in segmental sclerosis is unknown. The aim of this study is to investigate the effect of a MMP inhibitor, doxycycline, on glomeruloscleresis (GS) in renal ablation nephropathy and to evaluate the MMP-2, MMP-9, tissue inhibitors of matrix metalloproteinases (TIMP)-1, TIMP-2, and collagen type IV expressions. Materials and Methods: Fourteen of the 32 female Wistar albinos were 5/6 nephrectomised. Doxycyline was given to half of each group (40 mg/ kg/day total 28 days). After sacrification, the GS, MMP-2 and MMP-9 expressions, and TIMP-2 expressions were analyzed histopathologically. Proand active MMP-2 and -9 were analyzed by gelatin zymography. TIMP-1 and TIMP-2 were measured with the enzyme-linked immunoassay. Results: Doxycycline administration to the 5/6 nephrectomy group improved GS but did not inhibit glomerular MMP-9 or cortical pro- and active- MMP-2 and pro-MMP9 but increased TIMP-1 and TIMP-2 expression in all groups in cortical tissue. Type IV collagen was decreased in the groups where GS were increased. MMP-9 expression and GS were increased in all groups receiving doxycycline. Conclusion: We have demonstrated improved GS in renal ablation model by doxycycline administration but also doxycycline has an unexpected adverse effect. The effect of doxycycline on the expression of MMP-2 and -9 cannot explain the improvement in GS but increased cortical TIPM-1 and -2 may be an important contributing factor for the inhibition of MMPs. Other types of MMPs and TIMPs may be important. Accumulation of other types of collagen may be prominent in GS of ablation nephropathy.

Keywords

• Matrix Metalloproteinases
• Tissue Inhibitors of Matrix Metalloproteinases
• Doxycycline
• Renal Ablation Nephropathy

Doksisiklinin 5/6 Renal Ablasyon Nefropatisi ile Gelişen Glomeruloskleroza Etkisi

Abstract

Amaç: Matriks metalloproteinaz (MMP) inhibitörlerinin böbrek hastalıklarına faydalı etkisi daha önce yayınlanmıştır. Ancak segmental skleroza olan etkileri belirsizdir. Bu çalışmanın amacı bir MMP inhibitörü olan doksisiklinin renal ablasyon nefropatisi ile oluşturulmuş glomerüloskleroza (GS) etkisi ve MMP-2, MMP-9, matriks metalloproteinaz doku inhibitörleri (TIMP)-1, TIMP-2 ve kollajen tip IV ekspresyonlarını incelemektir. Gereç ve Yöntem: Çalışmamızda 32 adet dişi Wistar albino sıçanın 14’üne 5/6 nefrektomi uygulanmış; bunların ve nefrektomi uygulanmayanların yarısına oral doksisiklin verilmiştir (40 mg/kg/gün toplam 28 gün). Sakrifikasyon sonrası histopatolojik olarak glomerüllerdeki sklerotik değişikliklerin yanı sıra, immünfloresans ile MMP-9 düzeyleri ve immünhistokimyasal olarak TIMP-2 ekspresyonu değerlendirilmiştir. Ayrıca enzim bağlı immünosorbent deneyi ile, TIMP-1 ve TIMP-2 düzeyleri; jelatin zimografiyle pro ve aktif MMP-2 ve MMP-9 ölçülmüştür. Bulgular: Araştırma sonucunda 5/6 nefrektomi modelinde doksisiklinin glomerulosklerozu azaltıcı etkisi saptanmış ancak glomerular MMP-9 ya da kortikal pro ve aktif MMP-2 ve pro-MMP9 inhibisyonu görülmemiştir. Tüm gruplarda kortikal dokuda TIMP-1 ve TIMP-2 artışı saptanmıştır. Tip IV kollajen glomerulosklerozun arttığı gruplarda azalmıştır. MMP-9 ekspresyonu ve glomerüloskleroz doksisiklin alan tüm gruplarda azalmıştır. Sonuç: Doksisiklin uygulanması ile renal ablasyon modelinde glomerülosklerozun azaldığını gösterdik, ancak doksisiklinin beklenmedik bir yan etkisi görüldü. Doksisiklinin MMP-2 ve MMP-9 ekspresyonu etkisi glomerulosklerozdaki düzelmeyi açıklayamaz, fakat kortikal TIMP-1 ve TIMP-2’deki yükselme MMP inhibisyonundaki önemli bir faktör olabilir. Diğer MMP ve TIMP tipleri de önemli olabilir. Diğer kollajen tiplerindeki birikim ablasyon nefropatisinde meydana gelen glomerulosklerozda belirgin olabilir.

Keywords

• Matriks Metalloproteinaz
• Matriks Metalloproteinaz Doku İnhibitörleri
• Doksisiklin
• Reanal Ablasyon Nefropatisi

References

1. 1. Woessner JF Jr. Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J. 1991;5:2145-2154.
2. 2. Sarioglu S, Ozer E, Kirimca F, et al. Matrix metalloproteinases-2 expression in laryngeal preneoplastic and neoplastic lesions. Pathol Res Pract. 2001;197:483-486.
3. 3. Sis B, Sagol O, Küpelioglu A, et al. Prognostic significance of matrix metalloproteinases-2, cathepsin D, and tenascin-C expression in colorectal carcinoma. Pathol Res Pract. 2004;200:379-387.
4. 4. Ahuja TS. Doxycycline decreases proteinuria in glomerulonephritis. Am J Kidney Dis. 2003;42:376-380.
5. 5. Naini AE, Harandi AA, Moghtaderi J, et al. Doxycycline: a pilot study to reduce diabetic proteinuria. Am J Nephrol. 2007;27:269-273.
6. 6. Güray M, Sarioglu S, Türkmen M, et al. Cyclosporine A toxicity in association with reduced renal mass. Transplant Proc. 2003;35:3128-3133.
7. 7. Wu LL, Cox A, Roe CJ, et al. Transforming growth factor beta 1 and renal injury following subtotal nephrectomy in the rat: role of the reninangiotensin system. Kidney Int. 1997;51:1553-1567.
8. 8. Tatekawa Y, Kemmotsu H, Joe K, et al. Matrix Metalloproteinase-9 Expression in Congenital Diaphragmatic Hernia During Mechanical Ventilation. 2005;35:524–529.

9. 9. Sanders J, Goor H, Hanemaaijer R Renal expression of matrix metalloproteinases in human ANCA-associated glomerulonephritis., Nephrol Dial Transplant. 2004:1:1412-1419.
10. 10. Kleiner D, Stetler-Stevenson WG. Quantitative Zymography: Detection of Picogram Quantities of Gelatinases. Anal Biochem. 1994;1;218:325-329.
11. 11. Ahuja TS, Gopalani A, Davies P, et al. Matrix metalloproteinase-9 expression in renal biopsies of patients with HIV-associated nephropathy. Nephron Clin Pract. 2003;95:c100-c104.
12. 12. Liu S, Li Y, Zhao H, et al. Increase in extracellular cross-linking by tissue transglutaminase and reduction in expression of MMP-9 contribute differentially to focal segmental glomerulosclerosis in rats. Mol Cell Biochem. 2006;284::9-17.
13. 13. Johnson TS, Haylor JL, Thomas GL, et al. Matrix metalloproteinases and their inhibitions in experimental renal scarring. Exp Nephrol. 2002;10:182-195.
14. 14. Bauvois B, Mothu N, Nguyen J, et al. Specific changes in plasma concentrations of matrix metalloproteinase-2 and -9, TIMP-1 and TGFbeta1 in patients with distinct types of primary glomerulonephritis. Nephrol Dial Transplant. 2007;22:1115-1122.
15. 15. Ahmed AK, Haylor JL, El Nahas AM, et al. Localization of matrix metalloproteinases and their inhibitors in experimental progressive kidney scarring. Kidney Int. 2007;71:755-763.
16. 16. Cai YI, Sich M, Beziau A, et al. Collagen distribution in focal and segmental glomerulosclerosis: an immunofluorescence and ultrastructural immunogold study. J Pathol. 1996;179:188-196.
17. 17. Olgemöller B, Schleicher E. Alterations of glomerular matrix proteins in the pathogenesis of diabetic nephropathy. Clin Investig. 1993;71(5 Suppl):S13-S19.
18. 18. Floege J, Johnson RJ, Gordon K, et al. Increased synthesis of extracellular matrix mesengial proliferative nephritis. Kidney Int 1991;40:477-488.
19. 19. Munkert A, Helmchen U, Kemper MJ, et al. Characterization of the transcriptional regulation of the human MT1-MMP gene and association of risk reduction for focal-segmental glomerulosclerosis with two functional promoter SNPs. Nephrol Dial Transplant. 2009;24:735-742.
20. 20. Björklund M, Koivunen E. 2005 Gelatinase-mediated migration and invasion of cancer cells. Biochim Biophys Acta. 25;1755:37-69.
21. 21. Hanemaaijer R, Visser H, Koolwijk P, et al. Inhibition of MMP synthesis by doxycycline and chemically modified tetracyclines (CMTs) in human endothelial cells. Adv Dent Res. 1998;12:114-118.
22. 22. Golub LM, McNamara TF, Ryan ME, et al. Adjunctive treatment with subantimicrobial doses of doxycycline: effects on gingival fluid collagenase activity and attachment loss in adult periodontitis. J Clin Periodontol. 2000;28:146-156.
23. 23. Lutz J, Yao Y, Song E, et al. Inhibition of matrix metalloproteinases during chronic allograft nephropathy in rats.Transplantation. 2005; 27;79:655-661.