The Effects of Remote Ischemic Preconditioning and N-Acetylcysteine with Remote Ischemic Preconditioning in Rat Hepatic Ischemia Reperfusion Injury Model

Background. Remote ischemic preconditioning (RIP) and pharmacological preconditioning are the effective methods that can be used to prevent ischemia reperfusion (IR) injury. The aim of this study was to evaluate the effects of RIP and N-Acetylcysteine (NAC) with RIP in the rat hepatic IR injury model. Materials and Methods. 28 rats were divided into 4 groups. Group I (sham): only laparotomy was performed. Group II (IR): following 30 minutes of hepatic pedicle occlusion, 4 hours of reperfusion was performed. Group III (RIP + IR): following 3 cycles of RIP, hepatic IR was performed. Group IV (RIP + NAC + IR): following RIP and intraperitoneal administration of NAC (150 mg/kg), hepatic IR was performed. All the rats were sacrificed after blood samples were taken for the measurements of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels and liver was processed for conventional histopathology. Results. The hepatic histopathological injury scores of RIP + IR and RIP + NAC + IR groups were significantly lower than IR group (P = 0.006, P = 0.003, resp.). There were no significant differences in AST and ALT values between the IR, RIP + IR, and RIP + NAC + IR groups. Conclusions. In the present study, it was demonstrated histopathologically that RIP and RIP + NAC decreased hepatic IR injury significantly

The Immunohistochemical Evaluation of Kidney An Experimentally Induced Hypertensive And Diabetic Rat Model

AMAÇ: Bu çalışmada, deneysel olarak ayrı ayrı ve birlikte diyabet ve hipertansiyon modeli oluşturulan sıçanlarda, lipoik asidin hipertansif ve diyabetik böbrek üzerindeki tedavi edici/hasar önleyici etkisinin araştırılması amaçlandı. YÖNTEMLER: Çalışmamızda Wistar cinsi ratlar 8 gruba ayrıldı (n=7). I. grup; Kontrol, II. Grup; Diabetes Mellitus, III. grup; 5/6 Nefrektomi, IV. Grup; Lipoik asit, V. Grup; 5/6 Nefrektomi+Diyabet, VI. grup, Diyabet+Lipoik asit VII. Grup; 5/6 Nefrektomi+Lipoik asit ve VIII. Grup; 5/6Nefrektomi+Diyabet+Lipoik asit. Diyabet modeli 45mg/kg STZ enjeksiyonu ile oluşturuldu ve hipertansiyon modeli için 5/6 nefrektomi modeli uygulandı. dl-α-Lipoik asit 30mg/kg/gün olacak şekilde 8 hafta oral gavaj yöntemi ile deneklere uygulandı. Böbrek dokuları rutin ışık mikroskobik doku takip işlemlerinden geçirilip parafin bloklara gömüldü. İmmünohistokimyasal olarak AT1 (Anjiyotensin II tip I reseptörü), VEGF (Vasküler Endotelial Büyüme Faktörü) ve ET1 (Endotelin 1) antikorları işaretlendi. BULGULAR: Diyabet ve nefrektomi modellerinin ayrı ayrı ve birlikte uygulandığı deneysel gruplarda (Grup 2, 3, 5), glomerüloskleroz, mononükleer hücre infiltrasyonu, intersitisiyel fibrozis, damarlarda ve tübüllerde dilatasyon ve hiyalin materyal birikimi ile tübüler yapıların dejenerasyonu gözlendi. Aynı gruplarda tübülointersitisiyel ve glomerüler AT1 azalırken, VEGF ve ET1 artış göstermişti. Lipoik asit tedavi gruplarında ise AT1'de artış, VEGF ve ET1'de ise Kontrol ve Lipoik asit grubuna benzer şekilde azalma gözlendi. SONUÇ: Diyabet ve hipertansiyonun birlikte gözlenmesinin böbrek hasarının hızlı ilerlemesine neden olduğu, lipoik asidin bu hastalıklara karşı böbrek üzerinde tedavi edici etkisinin olduğu sonucuna varıldı. OBJECTIVE: In this study, experimental diabetes and nephrectomy have been applied separately and together in order to investigate possible therapeutic/damage prevention effects of Lipoic acid on hypertensive and diabetic rat kidneys. METHODS: Wistar rats were divided into 8 groups (n=7); Group 1; Control, Group 2; Diabetes Mellitus, Group 3; 5/6 Nephrectomy, Group 4; Diabetes Mellitus+5/6 Nephrectomy, Group 5; Lipoic acid administration, Group 6; Diabetes Mellitus+Lipoic acid, Group 7; 5/6 Nephrectomy+Lipoic acid, Group 8; Diabetes Mellitus+5/6 Nephrectomy+Lipoic acid groups respectively. Diabetes was formed by 45mg/kg STZ injection and for hypertension nephrectomy 5/6 model was applied. dl-α-Lipoic acid 30mg/kg/day was fed by oral gavage for 8 weeks. Kidney tissues were embedded into paraffin block after routine light microscopic preparation. AT1 (Angiotensinojen II type 1 receptor), VEGF (Vascular Endothelial Growth Factor) and ET1 (Endothelin) antibodies were labelled immunohistochemically in same group. RESULTS: In groups where diyabetes and nephrectomy were applied separately and together, glomerulosclerosis, mononuclear cell infiltration, interstitial fibrosis, vascular and tubular dilatation and hyalin deposition and degeneration of tubular structures were seen in glomerules. In the same group: tubulointerstitial and glomerular AT1 was decreased but VEGF and ET1 were increased. In Lipoic acid treatment groups, AT1 was increased and VEFG and ET1 were decreased similar to Control and Lipoic acid group. CONCLUSION: We have come to the conclusion that diabetes nd hypertension together increases the rate of renal injury and lipoic acid has therapeutic effect on kidney

The Effects of N-Acetylcysteine on Kidney Apoptosis in a Rat Intraabdominal Sepsis Model

Objective: The objective of this study was to investigate the effect of N-acetylcysteine (NAC) on kidney apoptosis using a rat intraabdominal sepsis model. Material and Methods: Rats were randomised into three study groups: sham (n=7), sepsis (n=7), and NAC (n=7) groups. In sham group, only laparotomy was performed, whereas in both sepsis and NAC groups, cecal ligation and perforation were done. After surgical process, in sham and sepsis groups, 1 mL saline was given once daily intraperitoneally for three Days, and in NAC group, 150 mg/kg NAC was given once daily intraperitoneally for three days. Six hours after the last dose, midline laparotomy was performed to all rats, and both kidneys were removed for biochemical and histopathological samplings. Findings from these tissues were compared based on malondialdehyde levels, structural changes in renal corpus and proximal tubules, mononuclear cell infiltration, erythrocyte extravasation and cysteinyl aspartate-specific proteinases (caspases)-3 immunoreactivity. Data were analysed using Kruskal-Wallis and Mann Witney-U tests. Results: Structural changes in proximal tubules, caspase-3 immunoreactivity and mononuclear cell infiltration and erythrocyte extravasation were significantly increased in sepsis group when compared to sham (p<0.05). Mononuclear cell infiltration and erythrocyte extravasation were significantly less in NAC group when compared to sepsis (p<0.05). Structural changes in interstitial space were increased in both sepsis and NAC groups when compared to sham (p<0.05). Conclusion: Although the findings of this study demonstrated an anti-inflammatory effect of NAC on kidneys when used during early sepsis, it was concluded that NAC as a free radical scavenger should be further studied for its potential effects on cell healing and prevention of apoptosis

The Effects of N-Acetylcysteine on Kidney Apoptosis in a Rat Intraabdominal Sepsis Model

Objective: The objective of this study was to investigate the effect of N-acetylcysteine (NAC) on kidney apoptosis using a rat intraabdominal sepsis model. Material and Methods: Rats were randomised into three study groups: sham (n=7), sepsis (n=7), and NAC (n=7) groups. In sham group, only laparotomy was performed, whereas in both sepsis and NAC groups, cecal ligation and perforation were done. After surgical process, in sham and sepsis groups, 1 mL saline was given once daily intraperitoneally for three Days, and in NAC group, 150 mg/kg NAC was given once daily intraperitoneally for three days. Six hours after the last dose, midline laparotomy was performed to all rats, and both kidneys were removed for biochemical and histopathological samplings. Findings from these tissues were compared based on malondialdehyde levels, structural changes in renal corpus and proximal tubules, mononuclear cell infiltration, erythrocyte extravasation and cysteinyl aspartate-specific proteinases (caspases)-3 immunoreactivity. Data were analysed using Kruskal-Wallis and Mann Witney-U tests. Results: Structural changes in proximal tubules, caspase-3 immunoreactivity and mononuclear cell infiltration and erythrocyte extravasation were significantly increased in sepsis group when compared to sham (p<0.05). Mononuclear cell infiltration and erythrocyte extravasation were significantly less in NAC group when compared to sepsis (p<0.05). Structural changes in interstitial space were increased in both sepsis and NAC groups when compared to sham (p<0.05). Conclusion: Although the findings of this study demonstrated an anti-inflammatory effect of NAC on kidneys when used during early sepsis, it was concluded that NAC as a free radical scavenger should be further studied for its potential effects on cell healing and prevention of apoptosis

İnsan göbek kordonu Wharton jölesinden mezenkimal kök hücrelerin eldesinde izolasyon yöntemlerinin ve antioksidatif kültür koşullarının etkisi

çoğaltılması rejeneratif tıp uygulamalarının ilk aşamalarından biridir. Terapötik amaçlar için in vitro hücre çoğaltmada, mezenkimal kök hücreleri (MKH)’ nin muhtemelen oksidatif stresi içeren erken yaşlanmaya hızla girmeleri önemli bir sorundur. Klinikte tedavide en yaygın kullanılan hücreler, kemik iliği MKH (Kİ-MKH) olmakla birlikte, göbek kordonu çevresinde yer alan Wharton jölesi MKH (WJ-MKH)’ nin invaziv olmayan şekilde elde edilebilmeleri, doku reddine yol açan proteinleri eksprese etmemeleri ve immünosupresif özellikleri nedeni ile Kİ-MKH’ nden daha kullanışlı olabilecekleri ileri sürülmektedir. Çalışmamız ile WJ-MKH’nin eldesi ve çoğaltılmasında kullanılan izolasyon yöntemlerinin etkinliğinin karşılaştırılması ve anti-oksidatif kültür koşullarının hücre canlılığı ve kök hücrelere özgü yüzey antijen ekspresyonlarına etkisinin belirlenmesi amaçlandı. Gereç ve Yöntem: Çalışmaya dahil etme kriterlerine uyan toplam 17 sağlıklı gebeden doğum sonrası kordon örnekleri alındı. Wharton jölesinden enzimatik ve eksplant yöntemi ile kök hücreler elde edilerek, hücre canlılıkları ve proliferasyon kapasiteleri karşılaştırıldı. Bulgular: Kök hücrelerin, osteoblastlara, kondrositlere ve adipositlere farklılaşma kapasiteleri immunohistokimyasal olarak gösterildi. Hücrelerin çoğaltılması sürecinde CD44, CD73, CD90, CD105 yüzey antijenlerinin dördüncü pasaja kadar anlamlı değişmeden eksprese edildiği saptandı. Antioksidan moleküller N-asetil sistein ve askorbik asit ilavesi ile sağlanan kültür koşullarının hücre canlılığı ve hücre yüzey antijen ekspresyonları üzerine etkisi gösterildi. Sonuç: Wharton jölesinden MKH’lerin izolasyonunda eksplant yönteminin yüzey antijen ifadeleri anlamlı değişmeksizin, daha kısa zaman diliminde elde edilen toplam hücre sayısı nedeni ile enzimatik yönteme göre daha avantajlı olduğu sonucuna varıldı

İSKEMİ - REPERFÜZYON MODELİNDE, BÖBREK KORTEKSİNDEKİ HİSTOPATOLOJİK DEĞİŞİKLİKLER VE ERİTROPOETİNİN KORUYUCU ETKİSİ

Eritropoetin (EPO), böbrekten salgılanan glikoprotein yapıda ve hematopoetik sistem üzerine etkili bir hormondur

PRİMER OLİGODENDROSİT KÜLTÜRÜNDE METAMFETAMİNE BAĞLI HÜCRE ÖLÜMÜ

Psikostümilan bir ilaç olan metamfetaminin nöroksit etkisi in vitto ve in vivo çalışmalarla kanıtlanmıştır

The Effects of Remote Ischemic Preconditioning and N-Acetylcysteine with Remote Ischemic Preconditioning in Rat Hepatic Ischemia Reperfusion Injury Model

Background. Remote ischemic preconditioning (RIP) and pharmacological preconditioning are the effective methods that can be used to prevent ischemia reperfusion (IR) injury. The aim of this study was to evaluate the effects of RIP and N-Acetylcysteine (NAC) with RIP in the rat hepatic IR injury model. Materials and Methods. 28 rats were divided into 4 groups. Group I (sham): only laparotomy was performed. Group II (IR): following 30 minutes of hepatic pedicle occlusion, 4 hours of reperfusion was performed. Group III (RIP + IR): following 3 cycles of RIP, hepatic IR was performed. Group IV (RIP + NAC + IR): following RIP and intraperitoneal administration of NAC (150 mg/kg), hepatic IR was performed. All the rats were sacrificed after blood samples were taken for the measurements of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels and liver was processed for conventional histopathology. Results. The hepatic histopathological injury scores of RIP + IR and RIP + NAC + IR groups were significantly lower than IR group (P = 0.006, P = 0.003, resp.). There were no significant differences in AST and ALT values between the IR, RIP + IR, and RIP + NAC + IR groups. Conclusions. In the present study, it was demonstrated histopathologically that RIP and RIP + NAC decreased hepatic IR injury significantly

The Effects of Dexmedetomidine on Secondary Acute Lung and Kidney Injuries in the Rat Model of Intra-Abdominal Sepsis

In the present study, the effects of dexmedetomidine on secondary lung and kidney injuries were studied in the rat model of intra-abdominal sepsis by immunohistological and biochemical examinations. We measured serum creatinine, kidney tissue malondialdehide and plasma neutrophil gelatinase-associated lipocalin levels. In order to evaluate tissue injury we determined kidney tissue mononuclear cell infiltration score, alveolar macrophage count, histological kidney and lung injury scores and kidney and lung tissue immunoreactivity scores. We demonstrated that dexmedetomidine attenuates sepsis-induced lung and kidney injuries and apoptosis in the rat model of sepsis. There is still need for comparative studies in order to determine the effects of dexmedetomidine on organ functions in early human sepsis

Ischemic preconditioning attenuates lipid peroxidation and apoptosis in the cecal ligation and puncture model of sepsis

Sepsis and septic shock are are among the major causes of mortality in intensive care units. The lung and kidney are the organs most affected by sepsis. Evidence exists that lipid peroxidation and apoptosis may be responsible for the high mortality due to sepsis. Ischemic preconditioning (IP) is a method for the protection of tissues and organs against ischemia/reperfusion injury by reducing reactive oxygen species levels, lipid peroxidation and apoptosis. In the present study, the effects of IP were investigated in cecal ligation and puncture (CLP)-induced sepsis in rats. The three groups of animals used in the present controlled study were the sham-operated group (sham, n=7), which only underwent a laparotomy; the sepsis group (sepsis, n=7), which underwent cecal ligation and perforation; and the IP + sepsis group (IP+sepsis, n=7), which underwent CLP immediately prior to the application of three cycles of IP to the hind limb. The study was terminated at 6 h after the induction of CLP. Blood, kidney and lung tissue samples were collected for the determination of serum creatinine, blood urea nitrogen (BUN), neutrophil gelatinase-associated lipocalin (NGAL) and lung tissue malondialdehyde (MDA) levels, as well as histological examination. The serum creatinine, plasma NGAL and lung tissue MDA levels in the sepsis group were significantly increased compared with those in the sham and the IP+sepsis groups (P<0.05). Alveolar macrophage counts, histological kidney and lung injury scores, kidney (caspase 3) and lung tissue immuonreactivity (M30) scores in the sepsis group were also significantly increased compared with those in the sham and IP+sepsis groups (P<0.05). The alveolar macrophage count in the IP+sepsis group was increased compared with that in the sham group (P<0.05). In conclusion, IP inhibits lipid peroxidation and attenuates histological injury and apoptosis in the lung and kidney during sepsis