Effect of chronic alcohol consumption on myocardial apoptosis in the rat model of isoproterenol-induced myocardial injury and investigation on the cardioprotective role of calpain inhibitor 1.

We investigated the presence of myocardial apoptosis on isoproterenol (ISO)-induced myocardial injury (MI) after long-term high dose alcohol consumption and examined the antiapoptotic role of calpain inhibitor 1. Male Wistar Albino rats (n = 108) were divided into six groups: Control, alcohol (ethanol was given during 30 days for chronic alcohol consumption), MI (150 mg/kg ISO injection at last two days of alcohol consumption), alcohol + MI, alcohol + MI + calpain inhibitor 1 (10 mg/kg inhibitor was injected at 15 min before ISO injections) and Dimethyl Sulfoxide (DMSO) groups. Biochemical, histological, and morphometric methods determined apoptosis levels in the heart tissue of rats. Cytochrome c, caspase 3, and calpain levels were significantly high in alcohol, MI, and alcohol + MI groups. In contrast, mitochondrial cardiolipin content was found to be low in alcohol, MI, and alcohol + MI groups. These parameters were close to the control group in the therapy group. Histological and morphometric data have supported biochemical results. As a result of our biochemical data, myocardial apoptosis was seen in the alcohol, MI, and especially alcohol after MI groups. Calpain inhibitor 1 reduced apoptotic cell death and prevented myocardial tissue injury in these groups. The efficiency of calpain inhibitor was very marked in MI after long-term high dose alcohol consumption.

Erythropoietin Protects the Kidney by Regulating the Effect of TNF-α in L-NAME-Induced Hypertensive Rats.

BACKGROUND/AIMS: Hypertension is the leading cause of death worldwide. Chronic high blood pressure induces inflammation. Tumor necrosis factor (TNF)-α plays a major role in inflammation and also depresses the synthesis of erythropoietin, which exerts protective effects on tissue; however, the mechanism is still unclear. We investigated the protective effect of erythropoietin against tissue damage caused by hypertension in the kidney and whether this effect was suppressed by TNF-α. METHODS: First, we detected the optimum chronic dose for darbepoetin-α (Depo), which is a long-acting erythropoietin analog for rats. We separated 60 female adult rats into 6 groups: control, Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), L-NAME+Depo, L-NAME+Remicade (an anti-TNF-α antibody), L-NAME+Depo+Remicade, Depo, and control. After 1 month of treatment, we measured cardiovascular parameters, took blood samples, sacrificed the rats, and removed kidneys for analyses. RESULTS: The apoptotic index and the plasma and kidney mRNA levels of TNF-α increased in the L-NAME group and decreased in all other treatment groups. Macrophage accumulation increased in the L-NAME and L-NAME+Remicade groups, while it decreased in the Depo group. The mRNA abundance of TNF receptor 1 (TNFR1) decreased slightly in the Depo group and TNFR2 increased significantly in the same group. CONCLUSION: Erythropoietin protects kidney tissue against hypertension by preventing the apoptotic effects of TNF-α by blocking macrophage accumulation, decreasing TNF-α levels, and switching the TNF-α receptors from the apoptotic receptor TNFR1 to the proliferative receptor TNFR2.

Effect of kefir and low-dose aspirin on arterial blood pressure measurements and renal apoptosis in unhypertensive rats with 4 weeks salt diet.

Abstract We aim to study the effect of low-dose aspirin and kefir on arterial blood pressure measurements and renal apoptosis in unhypertensive rats with 4 weeks salt diet. Forty adult male Sprague-Dawley rats were divided into five groups: control, high-salt (HS) (8.0% NaCl), HS+aspirin (10 mg/kg), HS+kefir (10.0%w/v), HS+aspirin +kefir. We measured sistolic blood pressure (SBP), mean arterial pressure (MAP), diastolic pressure, pulse pressure in the rats. Cathepsin B, L, DNA fragmentation and caspase-3 activities were determined from rat kidney tissues and rats clearance of creatinine calculated. Although HS diet increased significantly SBP, MAP, diastolic pressure, pulse pressure parameters compared the control values. They were not as high as accepted hypertension levels. When compared to HS groups, kefir groups significantly decrease Cathepsin B and DNA fragmentation levels. Caspase levels were elevated slightly in other groups according to control group. While, we also found that creatinine clearance was higher in HS+kefir and HS+low-dose aspirin than HS group. Thus, using low-dose aspirin had been approximately decreased of renal function damage. Kefir decreased renal function damage playing as Angiotensin-converting enzyme inhibitor. But, low-dose aspirin together with kefir worsened rat renal function damage. Cathepsin B might play role both apoptosis and prorenin-processing enzyme. But not caspase pathway may be involved in the present HS diet induced apoptosis. In conclusion, kefir and low-dose aspirin used independently protect renal function and renal damage induced by HS diet in rats.

Comparison of ankle force, mobility, flexibility, and plantar pressure values in athletes according to foot posture index.

Objectives: This study aims to compare ankle force, mobility, flexibility, and plantar pressure distribution of athletes according to foot posture index (FPI). Patients and methods: Between September 2016 and May 2018, a total of 70 volunteer male athletes (mean age: 21.1±2.3 years; range, 18 to 25 years) were included. The athletes were divided into three groups according to their FPI as follows: having supinated feet (Group 1, n=16), neutral/normal feet (Group 2, n=36), or pronated feet (Group 3, n=18). Ankle range of motion (ROM), muscle flexibility, ankle joint strength, and plantar pressure distribution were measured. Results: There were significant differences among the three groups in both right and left ankle dorsiflexion ROM (p=0.009 and p=0.003, respectively). Group 1 had significantly smaller dorsiflexion ROM than the other groups. Group 1 also showed significantly less flexibility in the gastrocnemius and soleus muscles than the other foot posture groups. Groups 2 and 3 exhibited significant differences in the maximum torque (p=0.018), maximum work (p=0.008), and total work (p=0.008) of the right plantar flexor muscles at 60°/sec angular velocity. Peak pressure measurements of the right foot were higher in Group 1, compared to Groups 2 and 3 (p<0.001). Conclusion: The results of this study may help to enhance athletic performance by providing a guide for designing training programs appropriate for athletes with different foot types to address their specific muscle flexibility and strength deficiencies.

Protective Effect of Boric Acid and Omega-3 on Myocardial Infarction in an Experimental Rat Model.

Boric acid and omega-3 are used as essential elements for both animal and human health. Many researchers have shown these beneficial effects on cardiac and inflammatory markers. This study aims to evaluate cardiac protective effect of boric acid and omega-3 against MI (myocardial infarction), probably due to the suppression of pro-inflammatory cytokines of natriuretic peptides in rats. Fifty male Sprague-Dawley rats were randomly divided into five groups: control, MI, MI+boric acid, MI+omega-3, and MI+boric acid+omega-3. Saline solution (2 ml/day), omega-3 (800 mg/kg/day), and boric acid (100 mg/kg/day)+omega-3 (800 mg/kg/day) were orally administered to the relevant groups throughout the 28 days. To constitute the MI model, the rats were exposed to isoproterenol-HCl (ISO) (200 mg/kg, S.C.) on the 27th and 28th. In the MI group, serum levels of CK-MB, BNP, and TNF-α are increased significantly. Also, ST waves and heart rates were higher in the MI than the control. These results demonstrate that biochemical results healed in MI+boric acid, MI+omega-3, and MI+boric acid+omega-3 groups compared MI group. ECG and light microscope results supported the findings as well. The statistical analysis showed that boric acid and/or omega-3 has protective effects on cellular damage in MI.

The protective effect of dexanabinol (HU-211) on nitric oxide and cysteine protease-mediated neuronal death in focal cerebral ischemia.

We hypothesized that dexanabinol can prevent neuronal death by protecting neuronal lysosomes from nitric oxide (NO)-mediated toxicity, and in turn, by suppressing the release of cathepsins during cerebral ischemia. Focal cerebral ischemia was induced in two sets of animals by permanent middle cerebral artery occlusion. The first set was used to monitor NO concentration and cathepsin activity, while the second was used for histological examination with hematoxylin and eosin, and TUNEL staining. In post-ischemic brain tissue, NO content and cathepsin B and L activity increased (p < 0.05). Dexanabinol treatment reduced NO concentration and cathepsin activity to the control level (p > 0.05). The number of eosinophilic and apoptotic neurons increased in the post-ischemic cerebral cortex (p < 0.05). However, dexanabinol treatment lowered both of these (p < 0.05). We conclude that dexanabinol might be a useful agent for the treatment of stroke patients.

Parenchymal Pressure Inconsistency in Different Brain Areas After Kaolin Injection into the Subarachnoid Space of Neonatal Rats.

AIM: To describe the relationship between the parenchymal pressure changes and the development of hydrocephalus in kaolininjected neonatal rats according to cerebral regions and time intervals of developing hydrocephalus. MATERIAL AND METHODS: Neonatal rats aged 2 to 3 days were examined in 5 groups as kaolin frontal "K-F", kaolin parietal "KP", saline frontal "SF-F", saline parietal "SF-P" and control "C", based on the injected material and injection sites. All injections were performed into the cortical subarachnoid space of the right frontal and right parietal regions. The fifth group was injection free. On the 3 < sup > rd < /sup > , 7 < sup > th < /sup > , 15 < sup > th < /sup > , 30 < sup > th < /sup > and 60 < sup > th < /sup > days after injection, parenchymal pressures (PP) of 5-7 rats from each group were measured from different regions. RESULTS: We compared the control group with saline-injected and kaolin-injected groups and found statistically significant parenchymal pressure differences based on regional measurements. In the kaolin groups, the mean PP values were obviously higher than the saline-injected group. Within each kaolin-injected group, the pressure values were variable and inconsistent regarding the parenchymal regions. CONCLUSION: Hydrocephalus cannot be totally explained with existent "bulk-flow" or "hydrodynamic" theories. Although our experimental design was planned to develop hydrocephalus according to the bulk flow theory, our results were more compatible with the hydrodynamic theory. The present comments on the occurrence and pathogenesis of hydrocephalus are still open to debate and may require further comprehensive studies.

Effect of vasoactive intestinal peptide on the wound healing of alkali-burned corneas.

AIM: To study the effect of vasoactive intestinal peptide (VIP) on wound healing in experimental alkali burns of the cornea. METHODS: Twenty-seven albino rabbits, weighing 3.2±0.75 kg were used. Alkali burns were induced on corneas by applying 10 mm Whatman paper No:50 soaked in 1 mol/L NaOH. They have further classified into 5 groups as follows: 1) control group given no treatment (n=5); 2) VIP given subconjunctivally (n=6); 3) VIP injected into anterior chamber (n=6); 4) NaCl 0.9% given subconjunctivally (n=5); 5) NaCl 0.9% given into the anterior chamber (n=5). All treatment protocols except control group were followed by topical eye drops composed of VIP at two hourly intervals for one week from 8 a.m. to 6 p.m. RESULTS: VIP treated groups of rabbits with alkali burns were found to have better wound healing findings histo-pathologically when compared to those of control group who have received no treatment on day 30. No differences were observed between groups in respect to degree of polymorphonuclear leukocytes (PMNL) infiltration and degree of loss of amorphous substrate on day 15. However, PMNL infiltration and degree of loss of amorphous substrate were lower in Groups 2 and 3 when compared to that of control group on day 30 (P<0.05). CONCLUSION: We have shown that VIP has positive effects on alkali induced corneal burns. VIP may inhibit PMNL migration to cornea through an immunomodulatory effect. Inhibition of PMNL migration might reduce the release of collagenases and this might prevent the extracellular amorphous substance loss.

Effect of quercetine and glutathione on the level of superoxide dismutase, catalase, malonyldialdehyde, blood pressure and neonatal outcome in a rat model of pre-eclampsia induced by NG-nitro-L-arginine-methyl ester.

OBJECTIVE: To anticipate the impact of antioxidant use on lipid peroxidation products, free oxygen radical scavengers, blood pressure (BP), proteinuria and neonatal outcome (as seen in percentage survival, litter birth weight) in a rat model of pre-eclampsia induced by NG-nitro-L-arginine-methyl-ester (L-NAME), an inhibitor of nitric oxide synthase (NOS). MATERIAL AND METHODS: Female adult non-pregnant Sprague-Dawley rats (n=40) with timed pregnancies were allocated into four groups according to medication they received on day 17 to term. Rats were randomised into a sham-treated group (group I, n=10) and groups treated with L-NAME, 50 mg/day i.p., only (group II, n=10), L-NAME + quercetine, 10mg/kg i.p. (group III, n=10) and L-NAME + glutathione, 60 mg/kg i.p. (group IV, n=10). Blood levels of superoxide dismutase (SOD), catalase (CAT) and malonyldialdehyde (MDA) were assessed on day 22 of gestation. Intracardiac blood sampling and hysterotomy were performed on day 22 of gestation. Mean systolic BP (measured with a tail-cuff device), level of proteinuria, total urine output, pups birth weight and percentages of live and of dead pups were recorded. RESULTS: Mean systolic BP and SOD, CAT and MDA levels were higher in rats infused with L-NAME than in the sham-treated group. In group IV, SOD levels were lower than in group II (P <0.001). A linear positive correlation between BPs on day 20 and SOD levels (rp=0.39) was recorded, as were negative correlations between level of proteinuria and SOD levels (rp=-0.39) and between CAT and MDA levels (rp=-0.39). Birth weights were higher in the sham-treated group than in the other groups (P <0.001). Pups of hypertensive gravid rats treated with antioxidants had better survival rates than those of rats in group II and the sham-treated group (Chi-square=15.9, d.f.: 3, P <0.01).However, no correlation was detected between higher pup mortality rate and birth weight of pups. CONCLUSION: In this rat model of pre-eclampsia, adverse outcomes, such as proteinuria and high neonatal death rate, are reversed by exogenous antioxidant use, even though no significant improvement is detected in terms of BP and birth weight of pups.

Gastroprotective effect of leptin on gastric mucosal injury induced by ischemia-reperfusion is related to gastric histamine content in rats.

Leptin has cytoprotective effect to gastric mucosal injury in rats. We aimed to test the hypothesis that leptin induced histamine is involved in the prevention of ischemia-reperfusion (I/R) induced gastric mucosal injury in rats. At the end of the 30 min celiac artery occlusion and 12h reperfusion process, serum and gastric tissue samples were taken from three group of rats to measure oxidative status, histamine levels and for histological examinations. Leptin decreased ulcer and polymorphonuclear leukocyte (PMNL) index, and serum malondialdehyde (MDA) and protein carbonyl content but increased gastric tissue histamine levels. We concluded that leptin exerts a protective effect on gastric mucosa to I/R induced gastric injury probably through increasing tissue histamine content which, in turn, maintain the gastric mucosal blood flow.

Erythropoietin prevents nitric oxide and cathepsin-mediated neuronal death in focal brain ischemia.

We examined the preventive effect of human recombinant erythropoietin (HrEPO) on nitric oxide (NO)-mediated toxicity to neurons and cysteine protease release into cytoplasm, which is attributed to neuronal death in brain ischemia. Focal cerebral ischemia was induced by permanent occlusion of middle cerebral artery in two sets of rat. The first set was used to monitor NO concentration and cathepsin activity, while the second was used for histological examination with hematoxylin and eosin, and TUNEL staining. A group in both set was administered human recombinant erythropoietin (HrEPO). NO content, cathepsins B and L activity increased significantly in the post-ischemic cerebral tissue (p<0.05). HrEPO treatment reduced NO concentration and cathepsin activity to control level (p>0.05). A significant increase in the number of necrotic and apoptotic neurons was observed in the post-ischemic cerebral cortex (p<0.05). HrEPO treatment was markedly lowered both of these (p<0.05). It is concluded that HrEPO prevents neuronal death by protecting neuronal liposomes from NO-mediated toxicity and suppressing the release of cathepsins.

Erythropoietin improves oxidative stress following spinal cord trauma in rats.

Spinal cord injury (SCI) is a very destructive process for both patients and society. Lipid peroxidation is the main cause of the further secondary damage which starts after mechanical destruction of tissues. Recent studies have shown that erythropoietin (EPO) has neuroprotective properties. In this study, we aimed to see the effect of EPO treatment after spinal cord injury on the oxidant and anti-oxidant enzyme systems and the relationship with the N-methyl-D-Aspartate (NMDA) blockage. Spinal cord injury was produced by epidural compression with a cerebral vascular clip that has a closing force of 40 g for 30s after a limited multilevel laminectomy (T9-11). Experiment was done in 5 groups: Group 1: Sham-operated untraumatised, Group 2: SCI untreated, Group 3: 150 i.u./kg EPO injected i.p. at the end of the first hour following the trauma. Group 4: NMDA receptor antagonist ketamine (100mg/kg) i.p. Group 5: EPO+ketamine i.p. The experiments were finished after 12h of the trauma. The spinal cords were excised for biochemical examinations. Anti-oxidant enzymes; catalase and reduced glutathione (GSH) levels increased and lipid peroxidation product, malonyldialdehyde (MDA) level decreased in EPO treated group when compared to the other groups. TNF-alpha levels decreased in EPO treated group. Application of ketamine before EPO treatment decreased effects of EPO. In conclusion, our results suggest that 150 i.u./kg i.p. EPO, a therapeutic dose in anaemic patients, applied after 1h of spinal cord injury significantly attenuated the oxidative damage of spinal cord injuries in rats. This activity is abolished via ketamine pretreatment.

Hypotensive effects of carvacrol on the blood pressure of normotensive rats.

Carvacrol is the major compound of essential oils of many plants, ethnomedically used for centuries but there were no detailed investigations on its action on the cardiovascular system. The aim of our study was to investigate the role of carvacrol on the cardiovascular functions of anesthetized rats and in vitro of isolated rat aorta. Carvacrol (100 microg/kg, I. P.) decreased heart rate, mean arterial pressure and systolic and diastolic blood pressures of the anesthetized rats whereas there were no effects at 1, 10 and 20 microg/kg. Carvacrol was observed to exhibit hypotension and to inhibit N((omega))-nitro- L-arginine methyl ester ( L-NAME)-induced hypertension. The lack of inhibitory action of carvacrol (10 (-4) M) on the CaCl (2)- and phenylephrine-induced contractions of isolated rat aorta showed that neither adrenergic receptors nor voltage-dependent vascular L-type calcium channels were involved. But, based on previous investigations, the involvement of cardiac L-type calcium channel blocking actions are suggested for the hypotensive actions of carvacrol was assumed.

Progressive ventricular enlargement in the absence of high ventricular pressure in an experimental neonatal rat model.

OBJECT: In the present study, we compared ventricular pressures (VP) and the progression of ventricular enlargement in a new experimental neonatal hydrocephalus model, to gain an understanding of how communicating hydrocephalus progresses. METHODS: Kaolin was injected into the subarachnoid space at the cranial convexity of neonatal rats. Gross examination was performed on the 3rd, 5th and 7th days, and ultrasonographic examination on the 15th day, and at the end of the 1st and 2nd months following the kaolin application. Ventricular size indexes (VSI) were calculated in the case of a large ventricular dilatation. VPs were assessed on the 15th day, and at the end of the 1st and 2nd months, with a computerized data acquisition system. CONCLUSIONS: In the 1st and 2nd months VSIs were significantly higher than in control rats on the 15th day after kaolin administration. VP on the 15th day was significantly increased compared with that in control rats. VP in the 1st month was still high, but had subsided. In the 2nd month VP was not increased over control. In the late stages, the progression of infantile communicating hydrocephalus is not related to VP levels.