Relationship of ozone application and time in rats with hypoxic-ischemic brain injury.

BACKGROUND: Despite the important advances in pregnancy and newborn follow-up, hypoxic-ischemic encephalopathy is still one of the prominent causes of newborn mortality and disability worldwide, and there is no sufficiently effective treatment for it yet. This study aimed to investigate whether the ozone injection, administered in a single-dose as a preconditioning agent before the hypoxia and in single and repeated doses on different days following the hypoxia, would affect the spatial memory performance of the rats in the Morris water maze test or on their apoptotic cell numbers. METHODS: The study consisted of 102 seven-day-old male Wistar baby rats randomly divided into five groups. Rats in all groups were induced with hypoxic-ischemic brain injury (HIBI) except for the Sham group, and 1.2 mg/kg ozone was administered intraperitoneally. For the apoptosis evaluation, eight rats from each of the first four groups were decapitated by cervical dislocation. Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay was used for immunohistochemical quantification of apoptosis in the excised brains. Blood malondialdehyde (MDA) and superoxide dismutase (SOD) levels were measured in the blood samples collected through cardiac puncture. Fourteen-week-old rats underwent the Morris water maze test to test their long-term spatial memory. RESULTS: On apoptotic quantification in the right hemisphere using the TUNEL assay, the numbers of apoptotic neurons in the ozone preconditioning group (Group 3) and the group given ozone on the day of hypoxia (Group 4) were found to be significantly higher than the Sham group (Group 1), but significantly lower than the non-treatment group (Group 2) (p <0.001; p <0.001, respectively). Group 3 rats had the highest mean MDA level and SOD activity. Considering the platform finding times in the first four days of the tests, Group 4 had the shortest times after Group 1; and on Day 4, Group 4 found the platforms significantly sooner than Groups 2, 3, and 5 (p <0.001). Comparison of Groups 1 and 4 revealed significantly shorter times for Group 1 for each day except for Day 2. CONCLUSIONS: Other studies have shown that controlled application of ozone would result in oxidative preconditioning and reduce the damage induced by reactive oxygen species through enabling adaptation to oxidative stress. Our study obtained remarkable and encouraging findings for ozone administration in HIBI by examining Group 4's performance in the first four days and the difference in its platform finding times between Day 1 and Day 4. HIPPOKRATIA 2021, 25 (2):56-62.

The efficacy of ozone therapy in neonatal rats with hypoxic ischemic brain injury.

OBJECTIVES: This study is aimed to determine the effect of ozone therapy in neonatal rats with experimentally induced hypoxic ischemic brain injury (HIBI). METHODS: The study included 7-d-old male Wistar rats that were randomized to the sham, control, ozone 1, and ozone 2 groups. All rats except those in the sham group were kept in a hypoxia chamber, and then the rats in the control group were given 0.5 mL of saline. Those in the ozone 1 group were given ozone 1 mg kg-1 intraperitoneally, and those in the ozone 2 group were given ozone 2 mg kg-1 intraperitoneally. RESULTS: There were significantly fewer apoptotic neurons in the right hemispheres of the rats in the ozone 1 and ozone 2 groups than in the control group (p < 0.001 and p < 0.001, respectively). There were significantly fewer apoptotic neurons in the right hemispheres of the rats in the ozone 2 group than in the ozone 1 group (p < 0.001). Morris Water Maze (MWM) test results were similar in the ozone 2 and sham groups. CONCLUSIONS: The present study's findings show that ozone therapy reduced neuronal apoptosis and improved cognitive function in neonatal rats with experimentally induced HIBI (Tab. 2, Ref. 30).

Is levetiracetam neuroprotective in neonatal rats with hypoxic ischemic brain injury?

BACKGROUND: The aim of this study was to determine if levetiracetam (LEV) is neuroprotective in neonatal rats with hypoxic-ischemic brain injury (HIBI). METHODS: The study included 7-d-old male Wistar rats that were randomly divided into the LEV400, LEV800, control, and sham groups. All the rats, except those in the sham group, underwent ligation of the carotid artery and were then kept in a hypoxic chamber containing 8% oxygen for 2 h. At the end of the hypoxic period the rats in the control group were administered saline solution 0.5 mL, the rats in the LEV400 group were administered LEV 400 mg.kg-1, and rats in the LEV800 group were administered LEV 800 mg.kg-1 via the intraperitoneal route. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) method was used to evaluate neuronal apoptosis in the rats. The Morris Water Maze (MWM) test was performed at age 14 weeks in order to evaluate cognitive function. RESULTS: The number of apoptotic neurons in the right hemispheres was significantly lower in the sham, LEV400, and LEV800 groups than in the control group (p < 0.001, p < 0.001, and p < 0.001, respectively). In addition, the number of apoptotic neurons in the right hemispheres was significantly lower in the LEV800 group than in the LEV400 group (p = 0.001). Platform finding time (PFT) during MWM testing was significantly shorter in the sham and LEV800 groups on d 4 than on d 1 (p = 0.001 and p = 0.006, respectively); however, PFT did not significantly change between d 1 and d 4 in the control or LEV400 groups (p = 0.91 and p = 0.096, respectively). CONCLUSION: Based on the present findings, LEV exhibited a dose-dependent neuroprotective effect in neonatal rats with HIBI (Ref. 27).