Medical ozone therapy reduces shock wave therapy-induced renal injury.

OBJECTIVES: Extracorporeal shock wave (ESW) lithotripsy is the preferred treatment modality for uncomplicated kidney stones. More recently free oxygen radical production following ESW application has been considered to be crucial in shock wave-induced renal damage. It has been shown that ozone therapy (OT) has ameliorative and preventive effects against various pathological conditions due to increased nitro-oxidative stress. In current study, we aimed to evaluate the efficacy of OT against ESW-induced renal injury. METHODS: Twenty-four male Sprague-Dawley rats were divided into three groups: sham-operated, ESW, and ESW + OT groups. All groups except sham-operated group were subjected to ESW procedure. ESW + OT group received 1 mg/kg/day of oxygen/ozone mixture intraperitoneally at 2 h before ESW, and OT was continued once a day for consecutive three days. The animals were killed at the 4th day, and kidney tissue and blood samples were harvested for biochemical and histopathologic analysis. RESULTS: Serum ALT and AST levels, serum neopterin, tissue nitrite/nitrate levels, and tissue oxidative stress parameters were increased in the ESW group and almost came close to control values in the treatment group (p < 0.05, ESW vs. ESW + OT). Histopathological injury scores were significantly lower in treatment group than the ESW group (p < 0.05, ESW vs. ESW + OT). Immunohistochemical iNOS staining scores in ESW group were higher than those of sham-operated group (p < 0.05, ESW vs. sham-operated), iNOS staining scores in OT group were significantly lower than the ESW group (p < 0.05, ESW + OT vs. ESW). CONCLUSION: OT ameliorates nitro-oxidative stress and reduces the severity of pathological changes in the experimental ESW-induced renal injury of rat model.

Efficacy of poly(adenosine diphosphate-ribose) polymerase inhibition in extracorporeal shock wave-induced renal injury.

OBJECTIVES: Extracorporeal shock wave lithotripsy (ESW) induces renal damage by excessive production of free oxygen radicals. Free Oxygen radicals cause cellular injury by inducing nicks in DNA. The enzyme poly(adenosine diphosphate-ribose) polymerase (PARP) involved in the process of repair of DNA in damaged cells. However, its activation in damaged cells can lead to adenosine triphosphate depletion and death. Thus, we designed a study to evaluate the efficacy of 3-aminobenzamide (3-AB), a PARP inhibitor, against extracorporeal shock wave induced renal injury. METHODS: Twenty-four Sprague-Dawley rats were divided into three groups: control, ESW, ESW + 3-AB groups. All groups except control group were subjected to ESW procedure. ESW + 3-AB group received 20 mg/kg/day 3-aminobenzamide intraperitoneally at 2 h before ESW and continued once a day for consecutive 3 days. The surviving animals were sacrificed at the 4th day and their kidneys were harvested for biochemical and histopathologic analysis. Blood samples from animals were also obtained. RESULTS: Serum ALT and AST levels, serum neopterin and tissue oxidative stress parameters were increased in the ESW group and almost came to control values in the treatment group (p < 0.05, ESW vs. ESW + 3-AB). Histopathological injury score were significantly lower in treatment group than the ESW group (p < 0.05, ESW vs. ESW + 3-AB). CONCLUSION: Our data showed that PARP inhibition protected renal tissue against ESW induced renal injury. These findings suggest that it would be possible to improve the outcome of ESW induced renal injury by using PARP inhibitors as a preventive therapy.

Inhibition of inducible nitric oxide synthase prevents shock wave therapy induced renal injury.

OBJECTIVES: Shock wave lithotripsy treatment (SWT) is not completely free from side effects; one of the accused mechanisms for renal injury during SWT is oxygen- and nitrogen-derived free radical productions. Therefore, we aimed to evaluate the effect of inhibition of nitric oxide (NO) production by N-[3(aminomethyl) benzyl) acetamidine] (1400W), highly selective inducible nitric oxide synthase (iNOS) inhibitor, at SWT-induced kidney damage. MATERIALS AND METHODS: Twenty-four rats those underwent right nephrectomy procedure were divided equally into three groups as control, SWT, and SWT + 1400W. 1400W was administered at a dose of 10 mg/kg at 2 h prior to SWT procedure and at the beginning of SWT procedure via intraperitoneal route and continued daily for consecutive 3 days. At the end of the fourth day, animals were killed via decapitation and trunk blood and the left kidneys were taken for biochemical and histopathologic evaluation. RESULTS: SWT caused renal tubular damage and increased lipid peroxidation and antioxidant enzyme activities and SWT also significantly increased nitro-oxidative products. Inhibition of iNOS via administration of 1400W ameliorated renal injury and decreased tissue lipid peroxidation (malondialdehyde), superoxide dismutase, glutathione peroxidase and nitrite/nitrate levels (NOx). In addition, it was seen that histolopathological changes were attenuated in the SWT + 1400W group when compared to SWT group. CONCLUSION: SWT-induced renal injury might be due to excessive production of oxygen free radicals and NO production. Inhibition of iNOS attenuates renal injury following SWT treatment. It can be concluded that iNOS inhibitors or peroxynitrite scavengers might be used to protect the kidneys against SWT-induced morphological and functional injuries.

The effects of N-acetylcysteine and ozone therapy on oxidative stress and inflammation in acetaminophen-induced nephrotoxicity model.

INTRODUCTION: Acetaminophen (APAP) is an analgesic and antipyretic agent. In overdoses, it is associated with nephrotoxicity. We examined the potential protective effects of N-acetylcysteine (NAC) and NAC + ozone therapy (OT) combination against APAP-induced nephrotoxicity. MATERIALS AND METHODS: Thirty-two male Sprague-Dawley rats were divided into four groups: sham, control (APAP), NAC, and NAC + OT. In the APAP, NAC, and NAC + OT groups, kidney injury was induced by oral administration of 1 g/kg APAP. The NAC group received NAC (100 mg/kg/day). NAC + OT group received NAC (100 mg/kg/day) and ozone/oxygen mixture (0.7 mg/kg/day) intraperitoneally for 5 days immediately after APAP administration. All animals were killed at 5 days after APAP administration. Renal tissues and blood samples were obtained for biochemical and histopathological analyses. Neopterin, tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-10 levels were measured in sera. Malondialdehyde (MDA) levels and glutathione peroxidase (GPx) activities were determined in renal homogenates. RESULTS: NAC and NAC + OT significantly decreased MDA and TNF-α levels and increased IL-10 levels and GPx activities. Serum neopterin and IL-6 levels were not different among all groups. APAP administration caused tubular necrosis in the kidney. The degrees of renal necrosis of the APAP group were higher than the other groups. Renal injury in rats treated with combination of NAC and OT were found to be significantly less than the other groups. CONCLUSIONS: Our results showed that NAC and OT prevented renal injury in rats and reduced inflammation. These findings suggest that combination of NAC and OT might improve renal damages because of both oxidative stress and inflammation.

The effects of medical ozone therapy on renal ischemia/reperfusion injury.

INTRODUCTION: This study was designed to investigate the possible beneficial effects of medical ozone therapy (OT), known as an immunomodulator and antioxidant, on the renal function, morphology, and biochemical parameters of oxidative stress in kidneys subjected to ischemia/reperfusion injury (IRI). MATERIALS AND METHODS: Thirty male Sprague-Dawley rats were classified into three groups: control, renal IRI, and renal IRI + OT. The IRI group was induced by bilateral renal ischemia for 60 min, followed by reperfusion for 6 h. After reperfusion, the kidneys and blood of rats were obtained for histopathologic and biochemical evaluation. RESULTS: Renal IRI increased the tissue oxidative stress parameters (lipid peroxidation, protein oxidation, and nitrite plus nitrate) and decreased the antioxidant enzyme activities (superoxide dismutase and glutathione peroxidase). The serum neopterin levels showed correlation with oxidative stress parameters. All these parameters were brought to control values in the treatment group. Histopathologically, the kidney injury in the treatment group was significantly lesser than in the renal IRI group. CONCLUSIONS: Our results clearly showed that OT has beneficial effect to protect kidney against IRI. The serum neopterin levels might be used as a marker to detect the degree of renal IRI.

Ozone therapy prevents renal inflammation and fibrosis in a rat model of acute pyelonephritis.

INTRODUCTION: Not only bacterial characteristics but also oxidative/nitrosative stress could play a significant role in renal parenchymal inflammatory processes in acute pyelonephritis (APN). This study was conducted to evaluate the effect of ozone therapy (OT), as an immunomodulator and antioxidant, on the renal function, morphology and biochemical parameters of oxidative stress in an experimental model of APN in rats. MATERIALS AND METHODS: Forty rats were divided equally into five groups as control, APN, APN + Antibiotic, APN + OT, and APN + Antibiotic + OT. APN was induced by 0.1 ml of freshly prepared Escherichia coli (ATCC 25922) solution containing 10(10) colony-forming unit/ml into the kidney. A control group was administered 0.1 ml of 0.9 % NaCl solution. Treatment was begun 72 h after bacterial inoculation. Control and APN groups were given 0.9% NaCl solution, APN + Antibiotic and APN + OT were given either antibiotic (ciprofloxacine 150 mg/kg intramuscular/twice daily) or OT. APN + Antibiotic + OT group was given both antibiotic and OT for five consecutive days. At the end of the seventh day, animals were killed via decapitation and trunk blood was collected. Both kidneys were harvested and one half of each kidney were immediately stored for antioxidant enzyme activity, tissue lipid peroxidation and protein carbonyl content. The remainder was fixed for histopathologic examination. RESULTS: E. coli-induced APN increased the renal glomerular and tubular dysfunction, oxidative stress parameters and antioxidant enzyme activities. Either antibiotherapy or OT markedly ameliorated renal dysfunction, the antioxidant status of the kidneys and histopathological injuries subjected to E. coli-induced APN. Interestingly, the combination of antibiotherapy and OT was much more effective than either of the treatment modalities alone. CONCLUSION: The combination of antibiotherapy and OT markedly ameliorated renal dysfunction and improved antioxidant status and histopathologic modalities in rats subjected to E. coli-induced APN than either antibiotherapy or OT treatment alone. Therefore, OT may be considered as an adjuvant therapy to classical antibiotherapy to prevent renal inflammation and fibrosis in APN.

Beneficial effects of N-acetylcysteine and ebselen on renal ischemia/reperfusion injury.

INTRODUCTION: It has been demonstrated that peroxynitrite accompanies acute renal ischemia and contributes to the pathophysiology of renal damage. Therefore, we aimed to investigate the roles of N-acetylcysteine (NAC), a well-known powerful antioxidant, and ebselen (E), a scavenger of peroxynitrite, on renal injury induced by renal ischemia/reperfusion injury (IRI) of rat kidney. MATERIALS AND METHODS: Forty male Sprague-Dawley rats were divided into five groups: sham, renal IRI, renal IRI+NAC, renal IRI+E, and renal IRI+NAC+E. IR injury was induced by 60 min of bilateral renal ischemia followed by 6 h of reperfusion. After reperfusion, kidneys and blood samples were obtained for histopathological and biochemical evaluations. RESULTS: Renal IR resulted in increased malondialdehyde and nitrite/nitrate levels suggesting increased lipid peroxidation and peroxynitrite production and decreased superoxide dismutase and glutathione peroxidase activities. Both NAC and E alone significantly decreased malondialdehyde and nitrite/nitrate levels and increased superoxide dismutase and glutathione peroxidase activities. Additionally in the renal IRI+NAC+E group, all biochemical results were quite close to those of sham group. Histopathologically, the kidney injury in rats treated with combination of NAC and E was found significantly less than the other groups. CONCLUSIONS: Both NAC and E are able to ameliorate IRI of the kidney by decreasing oxidative and nitrosative stresses and increasing free radical scavenger properties. Additionally, combination of NAC and E prevents kidney damage more than when each drug is used alone, suggesting that scavenging peroxynitrite nearby antioxidant activity is important in preventing renal IRI.

Effects of medical ozone therapy on acetaminophen-induced nephrotoxicity in rats.

Acetaminophen (APAP), also known as paracetamol, is the commonest cause of toxic ingestion in the world. Because overdose of APAP has life-threatening effects on kidney, treatment of APAP-induced nephrotoxicity has life-saving importance. Aim of the study was to evaluate the efficacy of medical ozone therapy in experimental model of APAP toxication. Twenty-one male Wistar rats (200-250 g) were randomly assigned into three groups containing seven rats each: Sham, control (only APAP treated), and APAP + ozone therapy groups. Rats were killed 48 hours after administration of APAP. Urea, creatinine levels in the blood, and malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activity in renal tissue were measured. Kidney tissues were stained with hematoxylin and eosin for histological assessment. APAP administration deteriorated the renal functions and significantly elevated renal MDA levels and depleted SOD and GSH-Px activities. Ozone therapy significantly reduced the MDA level, increased the SOD and GSH-Px activities, and normalized the renal histology. In conclusion, our study results are consistent with encouraging data for ozone therapy on APAP-induced nephrotoxicity in rats by improving antioxidant mechanism and oxidative stress.

The effect of iNOS inhibitors and hyperbaric oxygen treatment in a rat model of experimental colitis.

AIM: Our aim was to investigate the effectiveness of aminoguanidine (AMG), an inducible nitric oxide synthase inhibitor, and hyperbaric oxygen (HBO) treatment in an experimental colitis model. METHODS: We induced colitis in rats. In the control group, we applied 2 ml serum physiologic intraperitoneally for 7 days. In the HBO group, 100% oxygen at 2.4 atm pressure was applied for 7 days. In the AMG group, 100 mg/kg AMG was applied intraperitoneally for 7 days. In the HBO + AMG group, HBO and AMG were applied, respectively. At the end of 7 days, rats were sacrificed and the distal 10 cm part of colon was examined macro- and microscopically. RESULTS: Severity of colitis and NO activities were reduced by AMG, HBO, and HBO + AMG application. There was histologically significant improvement, especially in the HBO + AMG group. CONCLUSIONS: Both HBO and AMG were significantly effective in preventing weight loss, reducing NO activities, and severity of colitis, when comparing HBO and AMG separately.

Ameliorative effects of proanthocyanidin on renal ischemia/reperfusion injury.

INTRODUCTION: Several natural products have been reported to have beneficial effects on ischemia/reperfusion (I/R) injury, particularly from a preventative perspective. Therefore, this study was designed to investigate the efficiency of proanthocyanidin (PA), a natural product derived from grape seed, on renal dysfunction and injury induced by I/R of rat kidney. MATERIALS AND METHODS: Twenty-four male Sprague-Dawley rats were divided into three groups: sham-operated, I/R, I/R+PA. Rats were given PA (100 mg/kg/day peroral) 7 days prior to I/R. All rats except sham-operated underwent 60 min of bilateral renal ischemia followed by 6 h of reperfusion. After reperfusion, kidneys and blood were obtained for evaluation. Superoxide dismutase, glutathione peroxidase, malondialdehyde, protein carbonyl content, and nitrite/nitrate level (NO(x)) were determined in the renal tissue. Serum creatinine (S(Cr)), blood urea nitrogen (BUN), and aspartate aminotransferase (AST) were determined in the blood. Additionally, renal sections were used for histological grade of renal injury. RESULTS: PA significantly reduced the I/R-induced increases in S(Cr), BUN, and AST. In addition, PA markedly reduced elevated oxidative stress product, restored decreased antioxidant enzymes, and attenuated histological alterations. Moreover, PA attenuated the tissue NO(x), levels indicating reduced NO production. CONCLUSIONS: The pretreatment of rats with PA reduced the renal dysfunction and morphological changes, ameliorated cellular injury, and restored renal antioxidant enzymes caused by renal I/R.

Histological results of sclerotherapy, phototherapy, and pulsed 980-nm laser irradiation of the dorsal rabbit ear vein.

OBJECTIVE: To compare the histological changes occurring after three different treatment modalities for telangiectasias. MATERIALS AND METHODS: Thirty 16-week-old New Zealand white rabbits weighing 2.4-3.1 kg were enrolled in the study. The rabbits were divided into three groups. The group 1 received sclerotherapy, he group 2 received phototherapy, and group 3 received high-power diode laser treatments. All animals were treated on the right dorsal marginal ear vein. Biopsies were taken on days 1, 2, 7, and 30 post-treatment, and histopathogical evaluation was performed. RESULTS: Clinical and histological thrombosis occurred between days 1 and 7 in all groups. Superficial necrosis, neutrophil infiltration, and recanalization were mostly seen in group 3, whereas thrombosis was prominent in groups 1 and 2. CONCLUSIONS: All of the methods tested appear to have similar mechanisms of action, but had differing clinical and histological results. Phototherapy and laser treatment are non-invasive and do not require an exact, pinpoint technique, in contrast to sclerotherapy. However, sclerotherapy and phototherapy showed better results, especially with regard to recanalization.

Hyperbaric oxygen enhances the efficiency of 5-aminosalicylic acid in acetic acid-induced colitis in rats.

The aim of this study was to assess the efficiency of hyperbaric oxygen alone and in combination with 5-aminosalicylic acid in the acetic acid-induced colitis model, a well-known experimental model of inflammatory bowel disease in rats. Rats were randomly divided into five groups. In the noncolitis control group, rats were given isotonic saline, while in the other groups rats were treated by intracolonic administration of 4% acetic acid. In group 2, the untreated control group, no additional therapy was applied. In groups 3, 4, and 5 hyperbaric oxygen, 5-aminosalicylic acid. and 5-aminosalicylic acid + hyperbaric oxygen therapies were applied, respectively. Administration of acetic acid caused an inflammatory response in all animals. Histopathologic score was significantly higher in group 2 than in any other group. 5-Aminosalicylic acid and hyperbaric oxygen significantly decreased the histopathologic score (P < 0.05). Myeloperoxidase activity was also reduced significantly by 5-aminosalicylic acid (P < 0.05) but not by hyperbaric oxygen. The most prominent ameliorative effect, however, was seen in group 5 and the histopathologic score and myeloperoxidase activity were significantly lower than in groups 3 (P < 0.05) and 4 (P < 0.001). Hydroxyproline level also increased significantly in group 5, but not in groups 3 and 4 (P < 0.001). These findings indicate that hyperbaric oxygen therapy is effective in reducing the extent of colitis induced by acetic acid, although it is not as potent as 5-aminosalicylic acid. The combination of hyperbaric oxygen and 5-aminosalicylic acid, however, led to a much more prominent reduction in the severity of colitis. Hyperbaric oxygen may have a promising place in the treatment of inflammatory bowel disease.

Subacute cholestatic hepatitis likely related to the use of senna for chronic constipation.

We report a case of senna-induced cholestatic hepatitis which was not diagnosed at presentation. A 77 year old male was referred with abdominal pain, jaundice and elevated transaminase levels. A diagnosis of extrahepatic cholestasis was first suspected, due to the observation of a duodenal diverticulum and dilated proximal choledocus. However, the sphincterotomy did not improve cholestasis. At further evaluation, HBsAg was positive but serological work up was compatible with a healthy-carrier status. Further interrogation of the patient revealed a history of chronic senna intake to treat a chronic constipation. Liver biopsy showed bridging hepatocellular necrosis as well as canalicular cholestasis. Drug withdrawal resulted in a slow and progressive reduction in bilirubin levels and liver enzymes. In this case senna was likely the cause of a subacute cholestatic hepatitis exemplifying again the potential role of herbal related liver injury.

Effect of mesalazine, metronidazole and gentamicin on bacterial translocation in experimental colitis.

BACKGROUND AND AIM: In inflammatory bowel disease it has been established that enteric microorganisms are present in the final stage of the active inflammatory process. The purpose of the present study was to investigate the effects of mesalazine, and metronidazole-gentamicin combination, on bacterial translocation in an animal colitis model. METHODS: Fifty rats were stratified into five groups. The control group (group NC) was given only 2 mL saline enema and the remaining four groups were given 2 mL acetic acid enema. Group CC was the diseased control group. The treatment regimens started on the fifth day: mesalazine enema in group MesC, metronidazole-gentamicin in group MGC, and mesalazine + metronidazole + gentamicin in group MesMGC. After death on day 10, 2.5-cm colonic segments from all groups were weighed separately. In all rats, histopathological scoring was done, and samples from feces, blood, liver and spleen underwent microbiological analyses. RESULTS: For all diseased rats, both mean weight loss and colonic segment weight/bodyweight ratio was significantly higher than that in the sham group. As compared with other groups, body and colonic segment changes as well as histopathological scoring in rats receiving mesalazine enema either solely or in combination with the antibiotics were lower. No bacterial growth was found in the blood, liver and spleen of the rats in the control group while enteric bacteria, mainly Escherichia coli (35%) were the most common bacteria translocated to that in the latter. Antibiotic combination, alone or in combination with mesalazine was effective in reducing the bacterial translocation while mesalazine administration did not properly influence its regression. CONCLUSIONS: Gram-negative enteric bacteria, predominantly E. coli, was the most common bacteria isolated in bacterial translocation occurring in acetic acid-induced colitis. This trial showed that mesalazine alone did not incorporate the reduction of infectious events, despite its beneficial effect on inflammatory changes in experimental colitis. Metronidazole and gentamicin combination given intraperitoneally was more effective than topical mesalazine in decreasing bacterial translocation.