Ovarian Ischemia-reperfusion Research Articles

Prevention of infertility induced by ovarian ischemia reperfusion injury by benidipine in rats: Biochemical, gene expression, histopathological and immunohistochemical evaluation

IntroductionBenidipine has been reported to prevent the ischemia/reperfusion (I/R) damage in heart tissue and to suppress oxidant and proinflammatory cytokine production, increased by I/R. However, There was no information about the effects of benidipine on I/R injury in the ovary and the damage of I/R-induced infertility. ObjectivesThe aim of the study was to investigate the effects of benidipine on bilateral ovarian I/R injury and whether or not effective in the treatment of I/R-induced infertility in rats. MethodForty-eight females, albino Wistar rats were randomly divided into 4 groups: IRC group (ovarian I/R group, n=12), IRB-2 group (ovarian I/R+2mg/kg benidipine group, n=12), IRB-4 group (ovarian I/R+4mg/kg benidipine group, n=12) and HG group (healthy group with sham operation, n=12). In IRB-2 and IRB-4 groups, two hours ischemia and two hours reperfusion was performed following orally benidipine administration. After this I/R procedure, 6 rats from each group performed bilateral overectomy. Ovarian levels of malondialdehyde (MDA) and total glutathione (tGSH), ovarian gene expressions of interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) and also apoptosis were evaluated. The other 6 rats from each group were put in together with six male rats in separated cages for 2 months in order to reproduce. During this period, rats which did not become pregnant were accepted as infertile. ResultsMDA levels, expressions of TNF-α and IL-1β in IRC group were significantly higher than in the SGA group and tGSH was decreased. In total, 4mg/kg benidipine has better prevented ovaries from the increase of oxidants and proinflammatory cytokines, the decrease of antioxidants than 2mg/kg benedipine. In the histopathological examination hemorrhage, congestion, follicle degeneration, neutrophil infiltration and necrosis were seen in ovarian tissue of IRC group. Only dilated and congested blood vessels were found in the IRB-2 group. No histopathological finding was encountered in the IRB-4 group. I/R caused infertility in rats. In total, 4mg/kg benidipine prevented from infertility better than the dose of 2mg/kg benedipine. ConclusionIn total, 4mg/kg benidipine reduced I/R injury and I/R-related infertility more significantly compared to 2mg/kg benedipine in rat ovaries.

Antioxidant and antiapoptotic effects of erdosteine in a rat model of ovarian ischemia-reperfusion injury.

Objective(s):To evaluate the protective effect of erdosteine, an antiapoptotic and antioxidant agent, on torsion–detorsion evoked histopathological changes in experimental ovarian ischemia-reperfusion (IR) injury.Materials and Methods:Eighteen female Wistar albino rats were used in control, IR, and IR+Edosteine (IR-E) groups, (n=6 in each). The IR-E group received the erdosteine for seven days before the induction of torsion/retorsion, (10 mg/kg/days). The IR and IR-E groups were exposed to right unilateral adnexal torsion for 3 hr. Three hours later, re-laparotomy was performed, and the right ovaries were surgically excised. Oxidant and antioxidants levels were determined in serum. The ovarian tissue samples were received and fixed with 10% neutral buffered formalin. The sections were stained with H&E, anti-PCNA, and TUNEL.Results:The IR group were showed severe acute inflammation, polynuclear leukocytes and macrophages, stromal oedema and haemorrhage. Treatment with erdosteine in rats significantly retained degenerative changes in the ovary PCNA (+) cell numbers were significantly decreased in the IR and IR-E groups unlike the control group. However, its numbers were significantly increased in the IR-E group unlike the IR group. TUNEL (+) cell numbers were significantly increased in the IR group unlike the control and the IR-E groups. In erdosteine treated group, TUNEL (+) cells were detected significantly less than the IR group (P<0.05).Conclusion:In conclusion, erdosteine maybe a protective agent for ovarian damage and decreasing lipid peroxidation products and leukocytes aggregation after adnexal torsion in animals.

Protective effect of hydrogen rich saline solution on experimental ovarian ischemia reperfusion model in rats

BackgroundThe present study aimed to investigate the effects of hydrogen rich saline solution (HRSS) in a rat model of ovarian ischemia–reperfusion injury. MethodsThirty-six female Wistar-albino rats were grouped randomly, into six groups of six rats. The groups were classified as: sham (S), hydrogen (H), torsion (T), torsion/detorsion (TD), hydrogen-torsion (HT), and hydrogen-torsion/detorsion (HTD). Bilateral adnexal torsion was performed for 3h in all torsion groups. HRSS was given 5ml/kg in hydrogen groups intraperitoneally. Malondialdehyde (MDA) and glutathione-S-transferase (GST) levels were measured in both the plasma and tissue samples. Tissue sections were evaluated histopathologically, and the apoptotic index was detected by TUNEL assay. The results were analyzed by Kruskal–Wallis and Pearson chi-square tests using computer software, SPSS Version 20.0 for Windows. ResultsThe MDA levels were higher and GST levels were lower in the torsion and detorsion groups when compared to other groups, but the differences were insignificant (P>0.05). The MDA levels were lower and GST levels were higher in the HT and HTD groups compared with the T and TD groups (P>0.05). Follicular injury, edema, vascular congestion, loss of cohesion and apoptotic index were higher in the torsion groups but decreased in the groups that received HRSS. ConclusionsAccording to histopathological and biochemical examinations, HRSS is effective in attenuating ischemia–reperfusion induced ovary injury.

Effect of Kineret® on ovarian ischemia reperfusion injury in a rat model.

To investigate the effect of Kineret® on ischemia reperfusion (IR) injury in rat ovaries. Rats were divided into four groups: ovarian IR (IRG); 50 mg/kg Kineret® + ovarian IR (KIR-50); 100 mg/kg Kineret® + ovarian IR (KIR-100); and sham operation (SOC). KIR-50 (n = 10) and KIR-100 (n = 10) groups received an intraperitoneal injection of Kineret® at doses of 50 and 100 mg/kg, respectively. IRG and SOC (n = 10) rat groups were given distilled water as solvent using the same method. The results were compared between the groups. In rats in which IR occurred, oxidant parameters, such as malondialdehyde (MDA) and myeloperoxidase (MPO), were increased, the level of proinflammatory interleukin 1 beta (IL-1β) was elevated and total glutathione (tGSH) as an antioxidant was decreased in the ovarian tissues. Administration of Kineret® at a dose of 100 mg/kg inhibited the increase of MDA, MOP and IL-1β and a decrease in tGSH caused by IR more significantly than administration of Kineret® at a dose of 50 mg/kg. In addition, 100 mg/kg Kineret® significantly decreased severe hemorrhage, degeneration and inflammatory signs in the follicular cells, caused by IR. Kineret® at 100 mg/kg markedly ameliorated increased apoptosis in ovarian tissue with IR more significantly than 50 mg/kg kineret. Our findings indicate that Kineret® might be useful in clinical practice for the treatment of damage that may occur as a result of ovarian torsion.

The Ovarian Congestion after “U-74389G” Administration in Rats

Object: The possible recessing capacity of the antioxidant drug “U-74389G” was studied in a rat model. It included the evaluation of the mean Ovarian Congestion (OC) lesions after induced ovarian Ischemia-Reperfusion (IR) injury. Methods: The 40 used rats of mean mass 231.875 g were classified at 2 evaluation endpoints: one of 60 min for groups A and C and one of 120 min for groups B and D after reperfusion. The U-74389G was administered only in groups C and D. Results: U-74389G administration non-significantly declined the OC lesions scores by 0.6 mild [-1.208205–0.0082055] (p=0.0832). Reperfusion time non-significantly augmented the OC lesions scores by 0.4 without lesions [0.7678806–0.56788075] (p=0.2860). The interaction of U-74389G administration with reperfusion time non-significantly declined the OC lesions scores by 0.2727273 without lesions [-0.6477081–0.1022535] (p=0.1492). Conclusion: The U-74389G administration presented a no significant short-term recess direction for OC scores without lesions alteration. A more long-term study time or an enhanced U-74389G dose may outcome more significant effects.

Effects of curcumin on ovarian ischemia-reperfusion injury in a rat model.

Ischemia-reperfusion injury is a significant problem following reperfusion treatment for ovarian torsion. It is generally caused by reactive oxygen species-induced damage. Antioxidant agents, such as curcumin, may protect ovaries from this adverse effect. The aim of the present randomized, controlled study was to evaluate the short-term protective effect of curcumin on a rat model of ovarian ischemia-reperfusion injury. A total of 30 female Wistar albino rats, weighing 160-230 g, were divided into 2 groups depending upon the time of unilateral, left ovary ischemia/reperfusion (group 1, 2 h ischemia/2 h reperfusion and group 2, 4 h ischemia/4 h reperfusion). These groups were subdivided into 3 subgroups (sham, control and curcumin). The sham subgroups were not subjected to ischemia/reperfusion. Control and curcumin subgroups were performed under ischemia for 2 h plus 2 h reperfusion or 4 h ischemia plus 4 h reperfusion. Curcumin, 200 mg/kg, was intraperitoneally administered simultaneously with reperfusion to the curcumin subgroups. Serum nitric oxide (NO), NO synthase (NOS), xanthine oxidase (XO), total antioxidant status (TAS), total oxidant status (TOS) and histological scores were measured and compared between subgroups. For group 1, no significant differences were observed between NO, NOS, XO, TAS or TOS. The left ovary histological grade was significantly higher in the control and curcumin subgroups compared with the sham subgroup (P=0.036). For group 2, TOS was significantly higher in the control group compared with the sham and curcumin groups (P=0.023). However, TAS was also significantly higher in the control subgroup compared with the other 2 subgroups (P=0.005). Left ovary histological grade was significantly higher in the control and curcumin subgroups compared with the sham subgroup (P=0.038). No significant differences were observed between NO, NOS or XO between the group 2 subgroups. The results showed that curcumin exerted no major significant protective effect on ischemia-reperfusion injury in the rat ovary.

Octreotide protects ovary against ischemia-reperfusion injury in rats: Evaluation of histological and biochemical parameters.

This study investigated the efficacy of octreotide for prevention of ischemia-reperfusion injury in rat ovary. Thirty-two adult female rats were included. Rats were divided into five groups: in the sham group, the abdominal wall was only opened and closed; in the torsion group, ischemia was induced for 3 h using a torsion model involving atraumatic vascular clips; in the torsion/octreotide group, rats were given 100 µg/kg i.p. octreotide 30 min before torsion was induced; in the torsion/detorsion group, rats underwent 3 h ischemia-3 h reperfusion; in the torsion/detorsion/octreotide group, rats underwent 3 h ischemia followed by 100 µg/kg octreotide i.p. 30 min prior to 3 h reperfusion. Ovarian tissue damage was scored on histopathology. Ovarian tissue malondialdehyde and plasma pentraxin 3 were measured biochemically. In comparison with the sham group, both the torsion and torsion/detorsion groups had significantly higher scores for follicular degeneration, vascular congestion, edema, hemorrhage and leukocyte infiltration. Octreotide significantly decreased these scores in both groups. Ovarian malondialdehyde and plasma pentraxin 3 were significantly higher both in the torsion and torsion/detorsion groups compared with the sham group. Octreotide also decreased these levels significantly both in the torsion/octreotide and torsion/detorsion/octreotide groups. Octreotide ameliorated the potential side-effects of ovarian ischemia-reperfusion injury in a rat model.

The effects of tacrolimus on the activity and expression of tissue factor in the rat ovary with ischemia–reperfusion induced injury

In the present study, the effects of tacrolimus on the activity and expression of tissue factor (TF) were investigated in the ovarian ischemia–reperfusion induced injury in rats. Twenty-eight female rats (8–12 weeks, 300–350g) were divided into four groups: control, ischemia–reperfusion (IR), tacrolimus treated before ischemia (TBI), and tacrolimus treated before reperfusion (TBR) groups (n=7/per group). TF activity was measured using Quick's method, whereas TF expression was examined immunohistochemically. TF activity was significantly higher in all treated groups compared with the control group. Strong ovarian TF expression was demonstrated in the IR and TBR groups. Moreover, tacrolimus decreased TF activity in the TBI group compared with the IR group. The decreased activity of TF in the ovarian IR model may prevent IR-related inflammation during transplant procedure.

Protective effect of colchicine on ovarian ischemia-reperfusion injury: an experimental study.

The aim of the present study is to investigate the efficiency of colchicine in the experimental rat ovarian torsion model in the light of histological and biochemical data. A total of 35 Wistar albino female rats were randomly divided into 5 groups, group 1: (control-sham operated, n = 7); group 2: (torsion/detorsion, n = 7) 2 hours of ischemia and 2 hours of reperfusion; group 3: (torsion/detorsion, n = 7), 2 hours of ischemia and 5 days of reperfusion; group 4: (torsion/detorsion, n = 7) 2 hours of ischemia and 2 hours of reperfusion and a signal dose of oral 1 mL/kg colchicine; and group 5: (torsion/detorsion, n = 7), 2 hours of ischemia and 5 days of reperfusion and 5 days of oral 1 mg/kg colchicine. Histopathologic evaluation was performed by a scoring that assesses congestion, bleeding, edema, and cellular degeneration in the ovarian tissue. Catalase, tissue malondialdehyde (MDA), and protein carbonyl levels were calculated. The histopathologic scores, MDA, and protein carbonyl levels in the control and colchicine groups were significantly lower than groups 2 and 3 (P < .001). Catalase activities were significantly higher in the control and colchicine groups than in groups 2 and 3 (P < .001). The results of the histopathologic parameters and biochemical markers showed that protective effects of colchicine treatment persisted up to 5 days. Our study results revealed that colchicine reduced ovarian ischemia-reperfusion injury in experimental rat ovarian torsion model. As the ovarian detorsion is the first choice of the treatment modality in the early phase, antioxidant and anti-inflammatory treatment modalities like colchicine might be used to reduce ovarian ischemia-reperfusion injury.

Antioxidant effect of erdosteine and lipoic acid in ovarian ischemia–reperfusion injury

ObjectiveTo investigate the effects of erdosteine and alpha lipoic acid (ALA) in a rat model of ovarian ischaemia–reperfusion injury. Study designForty-eight female Wistar albino rats were separated, at random, into six groups of eight rats. The groups were classified as: sham, torsion, detorsion, detorsion+erdosteine 100mg/kg, detorsion+alpha lipoic acid (ALA) 100mg/kg, and detorsion+erdosteine+ALA. The investigators executing the biochemical and histological analyses were blinded to the randomization until the end of the study. ResultsThe TOS (Total Oxidant Status) and OSI (Oxidative Stress Index) levels are higher in the Torsion and Detorsion groups when compared with the ones in the Sham group (p<0.05). Strong correlation was found between OSI and total histological score in the sham, torsion and detorsion groups (r=0.765, p<0.001). The mean levels of TOS and OSI in the rats that received erdosteine and/or ALA were significantly lower compared with the sham, torsion and detorsion groups (p<0.05). Mean TOS and mean OSI were lower in the detorsion+erdosteine+ALA group compared with the detorsion+erdosteine and detorsion+ALA groups (p<0.05). In comparison with the detorsion group, the numbers of primordial follicles (p=0.006) and primary follicles (p=0.036) were increased in the groups that received erdosteine and/or ALA. ConclusionsErdosteine and ALA decreased ischaemia–reperfusion injury in an experimental rat ovarian torsion model; combination treatment had a greater effect than either agent alone. Treatment with erdosteine and/or ALA was found to preserve the loss of reproductive capacity normally observed after ovarian torsion.

The protective effect of quercetin on IMA levels and apoptosis in experimental ovarian ischemia-reperfusion injury

ObjectiveTo investigate the protective effect of quercetin (QE), an anti-inflammatory and anti-oxidant agent, on torsion–detorsion induced histopathological changes and blood IMA levels in experimental ovarian ischemia-reperfusion (IR) injury. Study designTwenty-four female Wistar rats were randomly divided into four groups in this study (n=6). Group I, (sham operation); Group II, torsion–detorsion plus saline (IR); Group III, torsion–detorsion plus solvent (dimethylsulfoxide: DMSO, IR+DMSO); Group IV, torsion–detorsion plus 15mg/kg/bw quercetin (IR+QE) injected intraperitoneally 30min prior to detorsion. After 3h of reperfusion, the right ovaries were removed surgically. The ovary tissue samples were fixed in 10% formalin solution for histopathological and immunohistochemical examination. Blood samples were obtained at the end of the procedures for each group of animals. ResultsOvarian sections in Groups II and III showed higher follicular cell degeneration, hemorrhage, vascular congestion and edema when compared with Group I. Administration of quercetin in rats significantly prevented degenerative changes in the ovary. Significantly less histopathological changes were found in Group IV compared with Groups II and III. Caspase-3 and TUNEL positive cells were detected in the ovarian surface, follicle epithelium, and stromal cells in all experimental groups, and there was a significant increase in Groups II and III compared with Group I (P<0.05). Treatment with quercetin decreased the number of caspase-3 and TUNEL positive cells. IR increased the ischemia modified albumin (IMA) levels in comparison to the sham group (1.06±0.10ABSU and 0.92±0.08ABSU, P<0.05). Quercetin administration before IR reduced the levels of IMA (0.93±0.08ABSU, P<0.05). ConclusionAdministration of quercetin is effective in preventing tissue damage induced by IR injury in ovaries.

Protective effect of oxytocin on ovarian ischemia-reperfusion injury in rats

Oxytocin (OT), a neurohypophysial nonapeptide, plays dual role as a neurotransmitter/neuromodulator and a hormone. It has also well known protective properties against ischemia/reperfusion organ damage. This study investigated the effect of OT on experimentally induced ovarian torsion/de-torsion ischemia/reperfusion (I/R) injury in rats. Sprague-Dawley rats were assigned to five treatment groups (n=7/group): Group 1, sham-operated; Group 2, torsion; Group 3, 80IU/kg of OT administration 30min prior to torsion; Group 4, torsion/de-torsion; and Group 5, torsion followed by 80IU/kg of OT administration 30min prior to de-torsion. OT administration significantly decreased the tissue malondialdehyde (MDA) levels in both the torsion and OT group (Group 3), and torsion/de-torsion OT group (Group 5) in comparison with the torsion-only group (Group 2) and torsion/de-torsion group (Group 4). Histopathological finding scores including follicular degeneration, edema, hemorrhage, vascular congestion, and infiltration by inflammatory cells were found to be significantly decreased in the torsion and OT group (Group 3), and torsion/de-torsion OT group (Group 5) when compared with the torsion-only group (Group 2) and torsion/de-torsion group (Group 4). In conclusion, these results, verified with histopathologic evaluation and biochemical assays, suggest a probable protective role for OT in ischemia and I/R injury in rat ovaries.

PSS68 - Diagnosis of Ovarian Ischemia-Reperfusion Injury via Optical Fiber Spectroscopy in a Rat Model

PSS68 - Diagnosis of Ovarian Ischemia-Reperfusion Injury via Optical Fiber Spectroscopy in a Rat Model

A comparative investigation of biochemical and histopathological effects of thiamine and thiamine pyrophosphate on ischemia–reperfusion induced oxidative damage in rat ovarian tissue

In this study, the biochemical and histopathological effects of thiamine and thiamine pyrophosphate on ischemia-reperfusion induced oxidative damage in rat ovarian tissue were investigated. Animals were divided into four groups of six rat each, ovarian ischemia-reperfusion (IR), 25 mg/kg thiamine + ovarian ischemia-reperfusion (TIR), 25 mg/kg thiamine pyrophosphate + ovarian ischemia-reperfusion (TPIR) and Sham group (SG). The results of the biochemical experiments have shown that the rat ovarian tissue with thiamine treatment, the level of MDA, GSH and the 8-hydroxyguanine are almost the same as the IR group; while in the group with thiamine pyrophosphate treatment, the level of MDA, GSH and the 8-hydroxyguanine are almost the same as the SG. Ovarian tissue of rats in the IR group were congested and dilated vessels, edema, hemorrhage, necrotic and apoptotic cells. In this group, the migration and the adhesion of the polymorphonuclear leucocytes to the endothelium were observed. Both ovaries in TPIR group, there was no difference according to the SG. Histopathology of ovarian tissues in the TIR group was almost the same with the IR group. Our results indicate that thiamine pyrophosphate significantly prevents the ischemia-reperfusion induced oxidative damage in ovarian tissue, whereas thiamine has no effect. In conclusion, we have found that thiamine pyrophosphate prevents oxidative damage due to ischemia-reperfusion injury, whereas thiamine does not have this effect. Furthermore, we have confirmed that the results of our biochemical analyses are in concordance with the histopathological findings.

The protective effect of erythropoietin and dimethylsulfoxide on ischemia-reperfusion injury in rat ovary

Objectives The aim of this study was to investigate the effects of erythropoietin and dimethylsulfoxide in the recovery from ischemia-reperfusion injury in an experimental rat adnexal torsion model. Study design Thirty-six Wistar-albino rats were divided into six groups. Except for the sham operation group, all groups were subjected to left unilateral adnexal torsion for 3 h. Erythropoietin and dimethylsulfoxide were intraperitoneally administered 30 min before the detorsion operation. Malondialdehyde and nitric oxide levels were detected from both the plasma and the tissue samples. The sections of the tissues were evaluated histologically. The results were analyzed by a one-way analysis of the variance (ANOVA) followed by the Duncan test for multiple comparisons using computer software, SPSS Version 15.0 for Windows. Results This study demonstrated that dimethylsulfoxide and erythropoietin pretreatment attenuated ischemia-reperfusion-induced lipid peroxidation, prevented post-ischemic ovarian injury and helped to maintain the ovarian morphology. Malondialdehyde levels of plasma and ovary were higher in the torsion and detorsion groups than the sham group. This showed that ischemia-reperfusion had caused lipid peroxidation of the ovarian tissue, thus leading to oxidative damage. One of the major findings of this study is that malondialdehyde was significantly decreased in the plasma of rats who were pre-treated with dimethylsulfoxide and erythropoietin before detorsion. This suggests that dimethylsulfoxide and erythropoietin might prevent oxidative damage in ovarian ischemia-reperfusion injury. Histological examination confirmed that reperfusion caused more detrimental effects than only ischemia, which could be at least partially prevented by dimethylsulfoxide and erythropoietin administration prior to detorsion. Conclusion Erythropoietin and dimethylsulfoxide may have beneficial effects in ischemia-reperfusion injury in ovarian torsion.

Effects of Pentoxifylline and Vitamin E on the Bilateral Ovary After Experimental Ovarian Ischemia

Pentoxifylline (PTX) and vitamin E inhibit the release of superoxide and hydroxyl radicals, and PTX improves capillary flow and tissue oxygenation. This experimental study was designed to determine the effects of PTX and vitamin E in the ovary after unilateral ovarian ischemia reperfusion (I-R) in albino Wistar rats. A vascular clamp was placed on the left ovary for 4 hours in all groups except for the control group. Following this, in the ischemia (I) group bilateral ovariectomy was performed. Saline, PTX, vitamin E, and PTX plus vitamin E were infused 30 min before reperfusion in the reperfusion (R), pentoxifylline (P), vitamin E (E), and pentoxifylline plus vitamin E (PE) groups, respectively. After 4 hours of reperfusion, the ovaries were removed for biochemical and histologic examination. MDA levels of bilateral ovaries in the PE group were significantly lower than in the E group (p < 0.0033). NO levels of bilateral ovaries in the PE group were significantly lower than in the P and E groups (p < 0.0033). Massive hemorrhage was determined in the ipsilateral ovaries of the R group. Hemorrhage was minimal or moderate in the ipsilateral ovaries of other groups. The contralateral ovaries showed congestion in different degrees. The contralateral ovaries of the group PE and the bilateral ovaries of the control group showed no pathological changes. PTX and vitamin E given together seems to be more effective in reducing I-R injury in ovarian tissue compared to administration of PTX, or vitamin E alone. However, further studies are required to evaluate the effective dose and duration of PTX and vitamin E on bilateral ovaries.