Matriks metalloproteinaz inhibitörü doksisiklinin enkapsüle peritoneal skleroz üzerine etkisi
Küçük Resim Yok
Tarih
2010
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
GİRİŞ: Peritonun sklerozu ve fibrozu periton diyalizi hastalarının en yıkıcıkomplikasyonu olan enkapsüle periton sklerozuna (EPS) yol açabilmektedir. Matriksmetalloproteinazların (MMP) aktivasyonu çeşitli inflamatuar ve fibrotik süreçlerekatkıda bulunmakta, anjiyogenez, hücre göçü, skleroz ve fibrozis sürecine öncülüketmektedir. Bu süreçte önemli olan, bazal membran gibi hücre dışı matrikselemanlarının MMP' ler tarafından yıkılmasıdır. Son zamanlarda MMP düzeylerininEPS gelişiminin erken bir laboratuar belirteci olabileceği bildirilmiştir. Bu çalışmada,bir MMP-2 inhibitörü olan doksisiklinin EPS üzerine etkilerinin deneysel rat modelindearaştırılması amaçlanmıştır.YÖNTEM: Kırk adet üremik olmayan Wistar Albino sıçan her grupta on hayvanolacak şekilde ayrıldı.Kontrol grubu: 0-3.haftalarda, günlük 2 mL %0.9 NaCl, intraperitoneal (İP) verildiKlorheksidin glukonat: (KG) grubu: 0-3.haftalarda, günlük 2 mL %0.1 KG ve %15etanol çözeltisi, İP verildi.Dinlenme grubu: 0-3.haftalarda, günlük 2 mL %0.1 KG ve %15 etanol çözeltisi, İPverildi. Ardından 4-6 haftalarda peritoneal dinlenmeye bırakıldı.Tedavi grubu (DOKS): 0-3.haftalarda, günlük 2 mL %0.1 KG ve %15 etanol çözeltisi,İP verildi. Ardından 4-6. haftalarda, 200 ?g/ml doksisiklin içeren içme sularıhazırlanarak tedavi verildi.Çalışmanın sonunda, bir saatlik periton eşitleme testi yapıldıktan sonra sıçanlarsakrifiye edilerek doğrudan kardiyak ponksiyon ile kan örneği, orta hat kesisi ile dediyalizat ve periton örnekleri alındı. Serum ve diyalizat biyokimyasal incelemeleri veperitonun patolojik değerlendirimi yapıldı. Diyalizat örneklerinde VEGF, MCP-1 veMMP-2 düzeyleri ELISA yöntemiyle ölçüldü. İstatistiksel analizler için Kruskal Wallistek yönlü ANOVA ve Mann Whitney U testi kullanıldı. p<0.05 anlamlı kabul edildi.SONUÇLAR: Üç hafta KG uygulaması periton membranında fonksiyonel ve yapısaldeğişikliklere yol açtı; visseral peritonda yaygın yapışıklıklar ve enkapsülasyonuneşlik ettiği deneysel EPS tablosu oluşturuldu.Üç hafta KG uygulandığında UF volümünde (27.3±0.3 ml'ye karşın 0.3±0.8 ml) veD1/D0 glukoz oranında (0.48±0.02'ye karşın 0.18±0.01) azalma saptandı (p<0.05).Bu azalma dinlenme ve doksisiklin grubunda anlamlı olarak düzeldi. D/P üre(0.51±0.02'ye karşın 0.82±0.03) ve D/P protein (18±2'ye karşın 54±7) oranlarındaartış saptandı (p<0.05). Bu artış doksisiklin grubunda anlamlı olarak geriledi. Diyalizatlökosit sayısında artış (739±47'ye karşın 1025±125/mm3) saptandı (p<0.05). Bu artışdinlenme grubunda kısmen düzelse de (727±99) doksisiklin grubunda (378±77)anlamlı olarak düşük bulundu (p<0.05). KG grubunda tüm diyalizat sitokin düzeylerikontrol grubuna göre artmıştı: VEGF (22±2'ye karşın 63±3), MCP-1 (34±3'e karşın444±28) ve MMP-2 (13±2'ye karşın 163±11). Dinlenme grubunda bu artışlardagerileme saptandı: (VEGF: 51±5), (MCP-1: 150±40) ve (MMP-2: 28±5) (p<0.05).Doksisiklin grubunda VEGF (26±3) ve MCP-1 (60±12) düzeyleri dinlenme grubunagöre düşük saptandı (p<0.05). MMP-2 (25±6) düzeyi doksisiklin grubunda dinlenmegrubuna göre düşük bulunsa da bu azalma istatistiksel anlama ulaşmadı.ÇIKARSAMA: Doksisiklin, peritonda oluşan fonksiyonel ve morfolojik değişiklikleri,inflamasyonu ve neovaskülarizasyonu azaltarak iyileştirmiştir. Doksisiklin EPStedavisinde yeni bir seçenek olabilir.Anahtar Kelimeler: doksisiklin, fibrozis, MMP-2, periton diyalizi
INTRODUCTION: Sclerosis and fibrosis of peritoneum ultimately may lead toencapsulating peritoneal sclerosis (EPS) which is the most devastating complicationin peritoneal dialysis patients. The activation of matrix metalloproteinases (MMPs,gelatinases) contributes to various inflammatory and fibrosing conditions. MMPs candegrade extracellular matrix components especially in basement membrane which isimportant for angiogenesis and migration of cells that promotes sclerosing andfibrosing process. More recently it has been shown that MMPs may be an earlydetection marker in EPS. Here in we present the effects of doxycycline, a MMPinhibitor, in experimental peritoneal sclerosing and fibrosing model.METHOD: We allocated forty non uremic wistar albino rats as follow:Control group: 2 mL isotonic saline intraperitoneally (IP) daily for 3 weeksCG group: 2 mL/200 g IP injection of 0.1% chlorhexidine gluconate (CG) and 15%ethanol dissolved in saline daily for a total of 3 weeksResting group: CG (weeks 0-3) plus peritoneal resting (weeks 4-6)Regression group: CG (0-3 weeks), plus doxycycline 200 ?Êg/ml in drinking water (4-6weeks).At the end of the study an one-hour peritoneal equilibration test was performed andrats were sacrified. Blood samples collected through direct cardiac puncture anddialysate samples through midline incision. Peritoneal functional and morphologicalchanges and dialysate VEGF, MCP-1 and MMP-2 levels were evaluated by EL.SA ineach groups.Kruskal Wallis, one way ANOVA and Mann Whitney U test were used for statisticalanalysis. p value < 0.05 was considered as statistically significant.RESULTS:Exposure to CG for 3 weeks resulted in alternations in peritoneal membran functionand morphology. Extensive adhesions in viceral peritoneum formed and experimentalEPS model created. Exposure to CG for 3 weeks; UF volume (27.3 }0.3 vs 0.3 }0.8ml) and D1/D0 glucouse (0.48 }0.02 vs 0.18 }0.01) decreased (p<0.05). This wassignificant in doxycycline and resting groups. D/P urea (0.51 }0.02 vs 0.82 }0.03) andD/P protein (18 }2 vs 54 }7) increased (p<0.05). In doxycycline group theseparameters were significantly reduced. Increase in dialysate cell count was detected(739 }47 vs 1025 }125/mm3) (p<0.05). This increase was partly ameliorated in restinggroup (727 }99) where as in doxycycline group (378 }77) this was statistically lowerthan CG group (p<0.05). Dialysate cytokine levels were increased in CG group thanother groups: VEGF (22 }2 vs 63 }3), MCP-1 (34 }3 vs 444 }28) and MMP-2 (13 }2 vs163 }11). In resting group significant decrease in these parametres was detected:(VEGF: 51 }5), (MCP-1: 150 }40) and (MMP-2: 28 }5) (p<0.05). In doxycycline grouplevels of VEGF (26 }3) and MCP-1 (60 }12) were lower than resting. MMP-2 (25 }6)level in doxycycline group was lower than resting group but did not reach statisticallysignificance.CONCLUSION: Doxycycline has significantly improved functional and structuralparameters of peritoneum by reducing inflammation and neovascularation.Doxycycline may be a future challenge in EPS.Key words: doxycycline, fibrosis, MMP-2, peritoneal dialysis
INTRODUCTION: Sclerosis and fibrosis of peritoneum ultimately may lead toencapsulating peritoneal sclerosis (EPS) which is the most devastating complicationin peritoneal dialysis patients. The activation of matrix metalloproteinases (MMPs,gelatinases) contributes to various inflammatory and fibrosing conditions. MMPs candegrade extracellular matrix components especially in basement membrane which isimportant for angiogenesis and migration of cells that promotes sclerosing andfibrosing process. More recently it has been shown that MMPs may be an earlydetection marker in EPS. Here in we present the effects of doxycycline, a MMPinhibitor, in experimental peritoneal sclerosing and fibrosing model.METHOD: We allocated forty non uremic wistar albino rats as follow:Control group: 2 mL isotonic saline intraperitoneally (IP) daily for 3 weeksCG group: 2 mL/200 g IP injection of 0.1% chlorhexidine gluconate (CG) and 15%ethanol dissolved in saline daily for a total of 3 weeksResting group: CG (weeks 0-3) plus peritoneal resting (weeks 4-6)Regression group: CG (0-3 weeks), plus doxycycline 200 ?Êg/ml in drinking water (4-6weeks).At the end of the study an one-hour peritoneal equilibration test was performed andrats were sacrified. Blood samples collected through direct cardiac puncture anddialysate samples through midline incision. Peritoneal functional and morphologicalchanges and dialysate VEGF, MCP-1 and MMP-2 levels were evaluated by EL.SA ineach groups.Kruskal Wallis, one way ANOVA and Mann Whitney U test were used for statisticalanalysis. p value < 0.05 was considered as statistically significant.RESULTS:Exposure to CG for 3 weeks resulted in alternations in peritoneal membran functionand morphology. Extensive adhesions in viceral peritoneum formed and experimentalEPS model created. Exposure to CG for 3 weeks; UF volume (27.3 }0.3 vs 0.3 }0.8ml) and D1/D0 glucouse (0.48 }0.02 vs 0.18 }0.01) decreased (p<0.05). This wassignificant in doxycycline and resting groups. D/P urea (0.51 }0.02 vs 0.82 }0.03) andD/P protein (18 }2 vs 54 }7) increased (p<0.05). In doxycycline group theseparameters were significantly reduced. Increase in dialysate cell count was detected(739 }47 vs 1025 }125/mm3) (p<0.05). This increase was partly ameliorated in restinggroup (727 }99) where as in doxycycline group (378 }77) this was statistically lowerthan CG group (p<0.05). Dialysate cytokine levels were increased in CG group thanother groups: VEGF (22 }2 vs 63 }3), MCP-1 (34 }3 vs 444 }28) and MMP-2 (13 }2 vs163 }11). In resting group significant decrease in these parametres was detected:(VEGF: 51 }5), (MCP-1: 150 }40) and (MMP-2: 28 }5) (p<0.05). In doxycycline grouplevels of VEGF (26 }3) and MCP-1 (60 }12) were lower than resting. MMP-2 (25 }6)level in doxycycline group was lower than resting group but did not reach statisticallysignificance.CONCLUSION: Doxycycline has significantly improved functional and structuralparameters of peritoneum by reducing inflammation and neovascularation.Doxycycline may be a future challenge in EPS.Key words: doxycycline, fibrosis, MMP-2, peritoneal dialysis
Açıklama
Anahtar Kelimeler
Fizyopatoloji, Physiopathology, Morfoloji, Morphology, Nefroloji, Nephrology, Böbrek yetmezliği-kronik, Kidney failure-chronic, Diyaliz, Dialysis, Doksisiklin, Doxycycline, Fibroz, Fibrosis, Matriks metalloproteinazlar, Matrix metalloproteinases, Periton diyalizi, Peritoneal dialysis, Skleroz, Sclerosis
