Morphological investigation of alcohol-induced hepatocyte apoptosis and liver injury in rats

The aim of this study was to morphologicall investigation of alcohol-induced hepatocyte apoptosis and liver injury in rats. A total of 20 male Sprague-Dawley rats were divided into two groups: control, and alcohol treated; each group contain 10 animals. The rats in alcohol treated group was given a daily dose of 6 g/kg ethanol by using intragastric intubation. Control group was given the same volume of saline. This application was continued daily for a total of 6 weeks. The end of the experiment all animals were anesthetized. The anesthetized rats were sacrificed and liver tissues were removed for histopathological investigation. Liver damage was examined by using hematoxylin-eosin and apoptosis was determined by terminal-deoxynucleotidyl- transferase mediated dUTP nick end labeling (TUNEL). There existed hepatocyte diffuse steatosis and hemorrhage in alcohol treated group. Our data indicate an enhancement in the activity of TUNEL in hepatocyte apoptosis of the alcholol treated group. The effects of alcohol on liver can be clearly detected as a hepatocyte cell death and liver injury. © 2011 OMU All rights reserved

Quercetin ameliorates methotrexate-induced renal damage, apoptosis and oxidative stress in rats

Background: In the present study, the protective and therapeutic effects of quercetin (QE) on renal injury induced by methotrexate (MTX) have been examined. Materials and methods: A total of 24 male rats were divided into the following three groups: control group, MTX group, and MTX+QE group. Rats in MTX group received 20mg/kg of single dose of MTX, while those in MTX+QE group received 20mg/kg of single dose MTX, in addition to 15mg/kg of QE administered 30min prior to MTX and in the following 5-day period as a single daily dose. At the end of the experimental period, renal tissues were removed for histopathological and biochemical assessments. Results: Light microscopic examination showed a disruption of the renal structure in rats in MTX group in the form of tubular degeneration and dilation, with shedding of the tubular epithelial cells into the lumen. QE treatment was associated with less marked degenerative changes, with a similar histological appearance to that of controls. Furthermore, QE treatment resulted in decreased the number of apoptotic cells. Biochemical assessments showed significantly higher malondialdehyde (MDA) levels in MTX group as compared to control and MTX+QE groups. superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) levels showed a significant decrease in MTX group as compared to controls. However, QE significantly suppressed MDA level, compensated deficits in the anti-oxidant defenses [reduced SOD, GSH-Px, and CAT levels] in kidney tissue resulted from MTX administration. Conclusions: In conclusion, renal toxic effects of MTX may be alleviated by QE

Deneysel Olarak Oluşturulan Testiküler İskemi Reperfüzyon Hasarına Karşı Urtica Dioica’Nın Koruyucu Etkisinin Apoptotik Ve Proliferatif Olarak İncelenmesi

Amaç: Testis torsiyonu çocukluk çağın akut skrotum tablosuna yol açan önemli sebeplerden biridir. Zamanında ve uygun tedavi edilmediği takdirde organ kaybıyla sonlanan ağır bir acil cerrahi problemdir. Testis torsiyonunun tedavisinde amaç İ/R’nin bozucu etkilerini azaltmak veya yok etmektir. Burada hücre membran bütünlüğünü koruyan etkili ajanlardan birisi antioksidanlardır. Bu çalışmada sıçanlarda deneysel testiküler İ/R modelinde antioksidan özellikleri olan Urtica dioica'nın (UD) antiapoptotik ve proliferatif olarak etkilerinin araştırılması amaçlandı. Gereç ve Yöntem: Çalışmamızda 24 adet erkek Sprague Dawley cinsi rat 3 gruba ayrılmıştır: Kontrol grubu, İ/R grubu ve İ/R+UD (2 mg/kg) grubu. Dört saat torsiyon ve 4 saatlik detorsiyon süresi sonunda sıçanlar anestezisi altında sakrifiye edildiler ve testis dokuları çıkartılarak histopatolojik değerlendirmeler yapıldı. Histopatolojik olarak STÇ ölçümü, Johnsen skorlaması, Hematoksilen-Eozin boyaması, proliferatif hücre çekirdek antijeni (PCNA) immünboyaması ve bir apoptozis belirteçi olan TUNEL boyaması yapıldı. Ayrıca tüm verilerin istatistiksel analizi ve gruplar arası karşılaştırmaları da yapıldı. Bulgular: Urtica dioica tedavisinin, ?/R sonrası dü?en STÇ ve JTBS’nu yükselttiği görüldü. İ/R sonrasında testislerin interstisyel yapısında bozulma, seminifer tübül yapılarında düzensizlik ve germ hücre dökülmesi gözlendi. Testisteki bu yapısal bozulmaların UD tedavisi sonrasında hafiflediği görüldü. Ayrıca UD tedavisinin İ/R sonucu azalan PCNA aktivitesini yükselttiği, artan TUNEL pozitif hücre sayısını ise düşürdüğü görüldü. Sonuç: Antiapoptotik ve koruyucu etkileri olan UD'nın, İ/R hasarının sıçan testislerinde neden olduğu histopatolojik değişiklikleri ve apoptozisi önlediği görüldü.Objective: Testicular torsion is one of the most important reasons that cause to acute scrotum table in childhood and it is an urgent surgical problem that result in losing the organ as long as not treating timely and appropriately. The aim is to reduce or eliminate the disruptive effects of ischemia reperfusion (?/R) in the treatment of the testicular torsion. Antioxidants are the one of the effective agents that protect the integrity of the cell membrane. In this study, it is aimed to examine the effects of urtica dioica (UD) that have antioxidant feature in the experimental testicular ?/R model in rats in terms of antiapoptatic and proliferative. Material and Method: In our study, 24 male rats have been divided into three groups: control group, ?/R group and ?/R+ UD (2 mg/ kg) group. At the end of the four hourly torsion and detorsion process, the rats have been sacrificed under anaesthesis and histopathological evaluations have been made by removing the testicular tissues. Seminiferous tubule caliber measurement, Johnson score, Haematoxylan-Eosin staining, proliferative cell nucleus antigen (PCNA) immunohistochemical staining and TUNEL as histopathological have been conducted Results: It has been observed that the treatment of UD has increased the Johnson score and seminife tubule caliber that decrease after ?/R, and deterioration in the interstitial structure of testicular, irregularities in the seminife tubule structure, and germ cell loss have been seen after ?/R. This structural deterioration in the testicular has reduced after the treatment of UD. It has also observed that the treatment of the UD has increased the activity of PCNA that decreases as a result of ?/R and this treatment has also reduced the number of the TUNEL positive cells that increase. Conclusion: It has observed that the protective effects of UD and histopathological changes in the testicular of the rat have been prevented the apoptosis and histological damage

Testicular tissues of experimental diabetic rats generated jnk and IL-6 relationship

Doktora TeziBu çalışmada, diyabetik erkek infertilitesinde rolü olabileceği ileri sürülen JNK’nın inhibisyonuyla veya inhibisyonsuz, hiperglisemiyle sinerjik bir etkileşimde bulunan IL-6 arasındaki ilişkinin ortaya konması amaçlanmıştır. Sprague Dawley erkek sıçanlar kullanılarak; kontrol (n=8), 2. grup (tek doz 60 mg/kg streptozotosin i.p.,15 gün;n=8), 3. grup (60 mg/kg streptozotosin,30 gün;n=8), 4. grup (60 mg/kg streptozotosin -15mg/kg SP600125 i.p.,15 gün;n=8) ve 5. grup (60 mg/kg streptozotosin -15mg/kg SP600125,30 gün;n=8) oluşturulmuştur. 2.,3.,4. ve 5. grupların testis/vücut ağırlık oranlarında,kontrol grubuna göre anlamlı bir azalma belirlenmiştir (P=0.001; P=0.002; P=0.006; P=0.011). Kontrol grubuyla karşılaştırılan, 2.,3.,4. ve 5. grup seminifer tübül çaplarının da azaldığı gözlenmiştir (sırasıyla P<0.001, P<0.001, P=0.012, P<0.001). 2. ile 4. grup ve 3. ile 5. gruplar karşılaştırıldığında ise inhibitör verilen gruplar lehine anlamlı bir yükselme (P<0.001) bulunmuştur. Tüm grupların Johnsen skor değerlerinin, kontrol grubuna göre anlamlı derecede azaldığı (P<0.001) ve 3.ile -5. gruplar karşılaştırıldığında ise 3. gruba göre anlamlı derecede artış (P=0.01) belirlenmiştir. Apoptotik tübül ve apoptotik hücre indeksleri kontrol grubu ile karşılaştırıldığında; 2.,3. ve 4. gruplarda anlamlı olarak artarken (2. ve 3. gruplar için P=0.021,P=0.020; 4. grup için P=0.043,P=0.020); 5. grupta ise 3. gruba göre apoptotik tübül (P=0.021) ve hücre (P=0.020) indeksleri azalmıştır. Bütün grupların fosfo- c-Jun N-terminal kinaz histolojik skorlama sonuçları incelendiğinde, kontrol grubu göre; 2. ve 3. grup değerlerinin yükseldiği (P=0.003,P=0.01); ancak SP600125 verilen 4. ve 5. gruplarda fark olmadığı görülmüştür (P=0.248,P=0,141). İnterlökin-6 immunreaktivitesinin ise 2.,3.,4. ve 5. gruplarda, kontrol grubuna göre yüksek olduğu (sırasıyla P=0.006,P=0.004, P= 0.010 ve 0.004), ancak 3. ile 5. gruplar kıyaslandığında, 5. gruptaki değerlerin anlamlı derecede azaldığı ( P=0.010) tespit edilmiştir.AbstractIn this study, it is aimed to introduce the relationship between IL-6 which is synergistically interacts with hyperglycemia and inhibition or without inhibition of JNK which is suggested to have a role in diabetic male fertility. Using Sprague-Dawley male rats, control (n=8), second group (single dose 60 mg/kg streptozotocin i.p.,15 days;n=8), third group (60 mg/kg streptozotocin,30 days;n=8), forth group (60 mg/kg streptozotocin –15mg/kg SP600125 i.p.,15 days;n=8) and fifth group (60 mg/kg streptozotocin -15 mg/kg SP600125,30 days;n=8) were assigned. In testis to body weight ratios of second,third,forth and fifth groups, significant increase was observed in comparison to control group (P=0.001; P=0.002; P=0.006; P=0.011). The seminiferous tubular diameter of second, third, forth and fifth groups decreased when it was compared to control group (P<0.001, P<0.001, P=0.012, P<0.001,respectively). When the groups were compared pairwise –second group with forth group and third group with fifth group, there was a significant increase in favour of inhibitor injected groups (P<0.001). The Johnsen scores of all groups decreased significantly comparing to control group (P<0.001) while third and fifth groups were compared, it was defined a significant increase in fifth group in contrast with third group (P=0.01). In consideration of apoptotic tubules and apoptotic cells indices; there is significant increase in second, third ve forth groups in comparison to control group( for second and third groups P=0.021,P=0.020; for forth group P=0.043,P=0.020); whereas there is a significant decrease in fifth group in apoptotic tubules(P=0.021) and cell (P=0.020) indices in comparison to third group. When phospho- c-Jun N-terminal kinaz HSCORE results of all groups were analyzed by comparison to control group; it was observed an increase in the values of second and third groups (P=0.003,P=0.01); but there is no difference in SP600125 injected forth and fifth gorups (P=0.248,P=0,141). The interleukin-6 immunoreactivity of second,third,forth ve fifth groups in comparison to control group was higher (P=0.006,P=0.004, P= 0.010 ve 0.004,respectively), but when third and fifth groups were compared, the values in fifth group was observed as decreased significantly( P=0.010)

MAPK immunoreactivity in streptozotocin-induced diabetic rat testis

PURPOSE: To evaluate the alterations of two mitogen-activated protein kinases (MAPK)s, extracellular signal regulated kinase (ERK) and c-Jun NH2 terminal kinase (JNK), in the testes of male rats with experimental diabetes.METHODS: Twenty males Sprague-Dawley rats were randomly divided into a control group (n=8) and a diabetes group (administration of 40 mg/kg/day streptozotocin (STZ) for five sequential days, n=12). After six weeks, testicular biopsy samples were obtained for light microscopy and immunohistochemical methods.RESULTS: The PCNA (proliferating cell nuclear antigen) index was significantly decreased in the diabetes group (p=0.004) when compared to the control group. Both total (t)-ERK and phosphor (p)-ERK immunoreactivities were significantly decreased in the diabetes group (p=0.004, p<0.001, respectively). The t-JNK immunoreactivity was unchanged in both groups (p=0.125), while p-JNK immunoreactivity was significantly increased in the diabetic group (p=0.002).CONCLUSIONS: The decrease of androgen levels in the course of diabetes may contribute to the decrease of the immunoreactivities of t-ERK and p-ERK. JNK may be activated due to the changes in various cytokines and chemochines that participate in the oxidative stress process of diabetes. Therefore, testicular apoptosis may occur and lead to infertility associated with diabetes. © 2014 Society for Neuroscience. All rights reserved

Effects of jnk inhibitor on inflammation and fibrosis in the ovary tissue of a rat model of polycystic ovary syndrome.

In our study, we aimed to investigate the effects of Jun N-terminal kinase inhibitor (SP600125) on fibrosis and inflammation in rats with polycystic ovary syndrome (PCOS)

Effects of jnk inhibitor on inflammation and fibrosis in the ovary tissue of a rat model of polycystic ovary syndrome

Objective: In our study, we aimed to investigate the effects of Jun N-terminal kinase inhibitor (SP600125) on fibrosis and inflammation in rats with polycystic ovary syndrome (PCOS). Method: 50 Wistar-albino rats were divided into five groups (n=10 each): control group, sham group, PCOS group, SP600125+ PCOS group and SP600125 group. In the estradiol valerate (EV)-treated group in which PCOS was injected with a single 4 mg/kg i.p. of EV in 0.2 ml sesame oil and the rats were sacrificed on day 60. The estradiol valerate (EV)-treated + SP600125-treated group was injected with a single 4 mg/kg i.p. of EV in 0.2 ml sesame oil. As of day 60, the treatment group was additionally given 15 mg/kg i.p. of SP600125 once daily for 4 consecutive days and the rats were sacrificed on day 65. Histopathological findings (ovarian morphology, edema, inflammatory cell infiltration, vascular congestion and hyperemia) and collagen type IV immunoexpression were assessed. Results: The SP600125+ PCOS group showed a significant level of improvement in ovarian follicle morphology, edema, inflammatory infiltrate, vascular congestion and hyperemia as compared with the PCOS group. Furthermore, collagen type IV immunoexpression showed a significant reduction in staining intensity on the theca cell layer and ovary stroma as compared to the PCOS group. Conclusion: This study demonstrates the therapeutic effect of SP600125 in the prevention of PCOS in an experimental model.Yuzuncu Yil University Scientific Research Project Coordination Unit, Van, TurkeyYuzuncu Yil University [2014-HIZ-TF079]This project is supported by a Grant from Yuzuncu Yil University Scientific Research Project Coordination Unit, Van, Turkey. (2014-HIZ-TF079)

Nigella sativa attenuates bleomycin-induced pulmonary fibrosis in rats by inhibition of inflammation, fibrosis, and inducible nitric oxide synthase

The present study investigated the anti-oxidant, anti-inflammatory and anti-fibrotic potential of Nigella sativa (NS) against bleomycin (BLC)-induced lung injury and fibrosis, an experimental animal model. A total of 18 male Sprague-Dawley rats were divided into three groups: Control, BLC, BLC+NS; each group consist of 6 animals. Pulmonary fibrosis was induced by a single intratracheal instillation of 2.5 mg/kg BLC. BLC+NS group rats were intragastric administered daily 400 mg/kg of NS from day 1 to 28. At the end of the study, lung tissues were removed for histopathological and immunohistochemical investigation. Lung tissues were stained with hematoxylin and eosin and Masson's Trichrome for histological evaluation. Moreover, the inducible nitric oxide synthase (iNOS) expression in the lung tissues was determined by immunohistochemical staining. BLC-induced histological changes including lung inflammation and lung fibrosis were significantly detected compared to the control group. NS treatment significantly ameliorated the BLC mediated histological changes and reduced the inflammatory cell infiltrate in lung tissues. NS significantly blocked collagen accumulations with parallel reduction in the fibrosis score. In addition, NS also markedly decreased the positive staining of iNOS in lung tissues. Our study provides evidence that NS significantly ameliorated BLM-induced pulmonary fibrosis in rats via the inhibition of inflammation score, fibrosis score, and iNOS expression. Therefore, NS may be a potential therapeutic reagent for the treatment of lung fibrosis. © 2015 OMU

Urtica dioica'nın sıçanlarda doksorubisine bağlı kardiyotosisitede histolojik hasar, oksidatif stress ve lipid peroksidasyon bakılayıcı etkileri]

Objective: Doxorubicin (DOX) is a highly effective anti-cancer drug with limited clinical use due to its serious cardiotoxicity. Urtica dioica L. seeds (UD), have been widely used in folk medicine, particularly in the therapy for advanced cancer patients, possesses a potent anti-oxidant properties. The goal of present study was to investigate the cardioprotective effects of UD on DOX-induced cardiotoxicity. Method: The rats in the UD treated group were given intraperitoneally 2 ml/kg UD. To induce cardiotoxicity, 30 mg/kg DOX was injected intraperitoneally by a single dose and the rats were sacrificed after 48 h. Results: The present study revealed for the first time a protective role of UD against DOX-induced cardiotoxicity. UD therapy significantly protected against DOX-induced myocardial damage which was characterized by conduction abnormalities, vacuolization, inflammatory cell infiltration, hemorrhages, and myofibrillar disarrangement. As indicators of oxidative stress, DOX caused significantly increase lipid peroxidation and reduction in activities of antioxidant enzymes; superoxide dismutase, glutathione peroxidase, and catalase. UD treatment significantly attenuated DOX-induced oxidative injury. Conclusion: The present study showed that UD might be a suitable cardioprotector against toxic effects of DOX

Protective effects of Urtica dioica L. on experimental testicular ischaemia reperfusion injury in rats

In this study, it was aimed to examine the effects of Urtica dioica L. (UD) that has antioxidant feature in the experimental testicular I/R model in rats in terms of anti-apoptotic and antioxidative effects. In our study, 24 male rats were divided into three groups: control group, I/R group and I/R+UD (2mgkg(-1)) group. Seminiferous tubule calibre measurement, Johnson score, haematoxylin-eosin staining, proliferative cell nucleus antigen (PCNA) immunohistochemical staining and TUNEL as histopathological have been conducted. The structural deterioration in the testicular on I/R group has reduced after the treatment of UD. Our data indicate a significant reduction in the activity of in situ identification of apoptosis using terminal dUTP nick end labelling (TUNEL), and there was a rise in the expression of proliferating cell nuclear antigen (PCNA) in testis tissues of UD-treated rats in the I/R group. The I/R+UD group showed a decrease in malondialdehyde levels and an increase in the activities of superoxide dismutase, catalase and glutathione peroxidase in comparison with the I/R group. It could be concluded that protective effects of UD on the I/R testicles are via reduction of histological damage, apoptosis, oxidative stress and lipid peroxidation.Namik Kemal University Research Center, Tekirdag, TurkeyNamik Kemal University [NKUBAP.00.20.AR.14.11]This study was supported under Project NKUBAP.00.20.AR.14.11 by the Namik Kemal University Research Center, Tekirdag, Turkey