Hypoxia conditioning could increase the survival of transplanted neuronal progenitor cells (NPCs) in rats with cerebral ischemia but could also hinder neuronal differentiation partly by suppressing mitochondrial metabolism. In this work, the mitochondrial metabolism of hypoxia-conditioned NPCs (hcNPCs) was upregulated via the additional administration of resveratrol, an herbal compound, to resolve the limitation of hypoxia conditioning on neuronal differentiation. Resveratrol was first applied during the in vitro neuronal differentiation of hcNPCs and concurrently promoted the differentiation, synaptogenesis, and functional development of neurons derived from hcNPCs and restored the mitochondrial metabolism. Furthermore, this herbal compound was used as an adjuvant during hcNPC transplantation in a photothrombotic stroke rat model. Resveratrol promoted neuronal differentiation and increased the long-term survival of transplanted hcNPCs. 18-fluorine fluorodeoxyglucose positron emission tomography and rotarod test showed that resveratrol and hcNPC transplantation synergistically improved the neurological and metabolic recovery of stroke rats. In conclusion, resveratrol promoted the neuronal differentiation and therapeutic efficiency of hcNPCs in stroke rats via restoring mitochondrial metabolism. This work suggested a novel approach to promote the clinical translation of NPC transplantation therapy.
Animals ; Brain Ischemia/drug therapy* ; Cell Differentiation ; Hypoxia ; Neurons ; Rats ; Resveratrol/pharmacology*

OBJECTIVEPharmacological intervention is an important means for the treatment of hypoxic-ischemic encephalopathy (HIE). As meta-analyses and randomized controlled clinical trials based on evidence-based medicine are able to provide the most reliable evidence for clinical practice, this study searched several databases in order to find the clinical evidence for the pharmacological treatment of neonatal HIE.

METHODSMeta-analyses and randomized (or quasi-randomized) controlled trials (RCT) for pharmacological therapy of HIE in term or late preterm newborn infants were searched from the databases of MEDLINE, EMBASE, Oxford Neonatal Group and Cochrane Library. The relevant literatures were statistically analyzed.

RESULTSFour Meta-analyses and thirteen RCTs were found to be involved in barbiturate, allopurinol, magnesium sulfate, mannitol, naloxone and dopamine therapy. None of the drugs could significantly decrease the mortality and the incidence of seizure or severe neurodevelopmental disabilities in newborn infants with HIE.

CONCLUSIONSCurrent clinical evidence indicates that no individual drug could decrease mortality and improve the neurodevelopmental outcomes in infants with HIE. Problems such as small scale in sampling and discrepancy in the identification of drug efficacy which exist in the clinical trials might lead to the uncertain effect of the drugs, and large sized and collaborative clinical trials are needed in the future.

Humans ; Hypoxia-Ischemia, Brain ; drug therapy ; mortality ; Infant, Newborn ; Randomized Controlled Trials as Topic

OBJECTIVE: To study the effect of different melatonin treatment regimens on long-term behavior and white matter damage in neonatal rats with hypoxic-ischemic brain damage (HIBD), and to seek an optimal melatonin treatment regimen. METHODS: Healthy Sprague-Dawley rats, aged 7 days, were randomly divided into four groups: sham-operation, HIBD, single-dose immediate treatment (SDIT), and 7-day continuous treatment (7DCT), with 8 rats in each group. A neonatal rat model of HIBD was prepared according to the classical Rice-Vannucci method. On day 21 after HIBD, the Morris water maze test was used to evaluate spatial learning and memory abilities. On day 70 after HIBD, immunofluorescence assay was used to measure the expression of neuronal nuclear antigen (NeuN) in the cerebral cortex and the hippocampal CA1 region of neonatal rats, and double-label immunofluorescence was used to measure the expression of myelin basic protein (MBP) and neurofilament 200 (NF200) in the corpus striatum and the corpus callosum. RESULTS: The results of the Morris water maze test showed that the SDIT and 7DCT groups had a significantly shorter mean escape latency than the HIBD group, and the 7DCT group had a significantly shorter mean escape latency than the SDIT group ( CONCLUSIONS Both SDIT and 7DCT can improve long-term behavior and reduce white matter damage in neonatal rats with HIBD, and 7DCT is more effective than SDIT.
Animals ; Animals, Newborn ; Hypoxia-Ischemia, Brain/drug therapy* ; Melatonin/pharmacology* ; Rats ; Rats, Sprague-Dawley ; White Matter

Neuroinflammation is a key contributor to the pathogenic cascades induced by hypoxic-ischemic (HI) insult in the neonatal brain. AD-16 is a novel anti-inflammatory compound, recently found to exert potent inhibition of the lipopolysaccharide-induced production of pro-inflammatory and neurotoxic mediators. In this study, we evaluated the effect of AD-16 on primary astrocytes and neurons under oxygen-glucose deprivation (OGD) in vitro and in mice with neonatal HI brain injury in vivo. We demonstrated that AD-16 protected against OGD-induced astrocytic and neuronal cell injury. Single dose post-treatment with AD-16 (1 mg/kg) improved the neurobehavioral outcome and reduced the infarct volume with a therapeutic window of up to 6 h. Chronic administration reduced the mortality rate and preserved whole-brain morphology following neonatal HI. The in vitro and in vivo effects suggest that AD-16 offers promising therapeutic efficacy in attenuating the progression of HI brain injury and protecting against the associated mortality and morbidity.
Animals ; Animals, Newborn ; Astrocytes/pathology* ; Brain/pathology* ; Brain Injuries/pathology* ; Glucose ; Hypoxia ; Hypoxia-Ischemia, Brain/drug therapy* ; Mice ; Neuroinflammatory Diseases ; Neuroprotective Agents/therapeutic use* ; Oxygen/therapeutic use*

OBJECTIVE: To study the effect of astragaloside IV (AS-IV) on NOD-like receptor protein 3 (NLRP3) inflammasome in neonatal rats with hypoxic-ischemic brain damage (HIBD). METHODS: A total of 24 Sprague-Dawley rats, aged 7 days, were randomly divided into a sham-operation group, an HIBD group, and an AS-IV treatment group, with 8 rats in each group. After 24 hours of modeling, brain tissue was collected for hematoxylin-eosin staining, yo-PRO-1 staining, and EthD-2 immunofluorescent staining in order to observe the cerebral protection effect of AS-IV in vivo. HT22 cells were used to prepare a model of oxygen-glycogen deprivation (OGD), and a concentration gradient (50-400 μmol/L) was established for AS-IV. CCK-8 assay was used to measure the viability of HT22 cells. RT-PCR and Western blot were used to observe the effect of different concentrations of AS-IV on the mRNA and protein expression of NLRP3, gasdermin D (GSDMD), caspase-1, and interleukin-1β (IL-1β). RESULTS: Yo-Pro-1 and EthD-2 staining showed that compared with the sham-operation group, the HIBD group had an increase in pyroptotic cells with a small number of necrotic cells, and the AS-IV group had reductions in both pyroptotic and necrotic cells. Compared with the sham-operation group, the HIBD group had significantly higher protein expression levels of NLRP3, IL-1β, caspase-1, and GSDMD ( CONCLUSIONS AS-IV may alleviate HIBD in neonatal rats by inhibiting the expression of NLRP3, GSDMD, caspase-1, and IL-1β.
Animals ; Animals, Newborn ; Brain ; Hypoxia-Ischemia, Brain/drug therapy* ; Inflammasomes ; NLR Proteins ; Rats ; Rats, Sprague-Dawley ; Saponins ; Triterpenes

Brain hypoxia-ischemia has been considered as critical factors in many human central nervous system diseases, including stroke and neonatal hypoxic-ischemic encephalopathy. In brain hypoxia-ischemia processes, inducible NO synthase (iNOS) is induced to produce excessive nitric oxide (NO) which leads to cascade reactions of inflammation and neuronal death, deteriorating primary brain injury. Inhibiting iNOS expression has opened new perspectives in the treatment of brain hypoxia-ischemia because iNOS inhibitor has been shown as a potent therapeutic agent. This reviews focus on recent research achievements regarding the relationship between iNOS and ischemic-hypoxic brain damage and the perspective of using iNOS inhibitors as therapeutic strategies for brain ischemic-hypoxic brain damage.
Animals ; Humans ; Hypoxia-Ischemia, Brain ; drug therapy ; metabolism ; Nitric Oxide Synthase Type II ; antagonists & inhibitors ; genetics ; physiology

OBJECTIVETo establish a model of immature rat hypoxic-ischemic brain damage (HIBD) which was expected to be similar to periventricular leukomalacia in human preterm infants pathologically and neuroethologically, and to investigate the role of minocycline (MN) in this model.

METHODTotally 192 Sprague-Dawley rats (postnatal day 2, P(2)), of either sex, were randomly divided into 4 groups: normal-group, sham operation group, HIBD-group, HIBD + MN group, each group had 48 rats. HIBD group and HIBD + MN group survived the left common carotid artery (CCA) ligation followed by 4h exposure to 8% O(2). Rats in sham operation group only survived the left CCA isolation. Rats in normal group were not treated with anything. In HIBD + MN group, the rats were treated with intraperitoneal injection of minocycline 45 mg/kg, immediately after HI and every 24 h for 2 days. Brain tissues were collected on day 3, 1 week, 2 weeks, 4 weeks after HI, for hematoxylin-eosin staining and histological scoring. Frozen sections of the brains were stained with anti-O4, anti-O1 immunohistochemistry on day 3 after HI, and MBP immunohistochemistry 2 weeks after HI. Rats in the four groups underwent neuroethologic examination 4 weeks after HI.

RESULTIn the HIBD group, there were pathological changes in the periventricular white matter. The pathological changes were milder in HIBD + MN group; There was no statistically significant difference between the normal group and HIBD + MN group in the number of positively stained O4 cell (P > 0.05). The number of positively stained O4 cell in the HIBD group was significantly reduced, compared with that of normal group, sham operation group, and HIBD + MN group (23.67 ± 12.00 vs. 52.89 ± 10.68, 39.28 ± 11.78, 41.63 ± 8.41, P < 0.05). The differences in the number of positively stained O1 cell among the normal group, sham operation group, HIBD group and HIBD + MN group had no statistical significance (P = 0.093). The numbers of myelin basic protein (MBP) positively immunostained fiber bundles in the HIBD + MN group were significantly less than that of the normal group and sham operation group (P < 0.05). The numbers of MBP positively immunostained fiber bundles in the HIBD group were significantly less than that of the normal group, sham operation group, and HIBD + MN group (14.71 ± 7.42 vs. 36.67 ± 6.50, 35.50 ± 3.24, 26.33 ± 5.92, P < 0.05). The HIBD group had long-term neuroethologic abnormality. There was no statistically significant difference in the inclined plane test, hanging test and cylinder test among the HIBD + MN group, normal group, and sham operation group (P > 0.05). The scores of the HIBD group had statistical significantly among the normal group, sham operation group and HIBD + MN group (P < 0.05). In the open field test, there was no statistically significant difference between the HIBD group and HIBD + MN group (P = 0.772), but there was significant difference between these two groups and the normal group, sham operation group (P < 0.05).

CONCLUSIONMinocycline protects the pre-oligodendrocyte and has protective effects in terms of long-term neuroethology.

Animals ; Animals, Newborn ; Disease Models, Animal ; Hypoxia-Ischemia, Brain ; drug therapy ; Minocycline ; therapeutic use ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the efficacy of erythropoietinin (EPO) in the treatment of moderate or severe hypoxic-ischemic encephalopathy (HIE) in neonates.

METHODSSeventy neonates with moderate or severe HIE were randomly assigned to two groups: EPO treatment and control (n=35 each). The EPO treatment group included 22 cases of moderate HIE and 13 cases of severe HIE. The control group included 24 cases of moderate HIE and 11 cases of severe HIE. Thirty-five healthy full-term infants served as normal group. The control group received a conventional treatment. Beside the conventional treatment, the EPO treatment group was intravenously injected with EPO of 200 IU/kg•d, 3 times weekly. Routine blood test was performed every 6 days. EPO dose was adjusted based on the results of the routine blood test. The course of EPO treatment was 2 to 4 weeks. Neonatal Behavioral Neurological Assessment (NBNA) was performed at age of 28 days. The infant development test of Child Development Centre of China (CDCC) was performed at ages of 3 months and 6 months.

RESULTSThe percentage of normal NBNA scores in the EPO treatment group was significantly higher than that in the control group at age of 28 days (P<0.05), but was significantly lower than that in the normal group (P<0.01). The CDCC test including physical development index (PDI) and physical development index (MDI) showed the percentage of normal results in the EPO treatment group was significantly higher than in the control group at age of 3 months (P<0.05), but was significantly lower than in the normal group (P<0.01). The CDCC test including PDI and MDI showed that the percentage of normal results in the EPO treatment group was significantly higher than in the control group at age of 6 months. The MDI test results in the EPO treatment group were not significantly different from those in the normal group at age of 6 months, but the percentage of normal results in the PDI test in the EPO treatment group was still significantly lower than that in the normal group (P<0.05).

CONCLUSIONSEPO treatment has neuroprotective effects against moderate or severe HIE and improves long-term behavioral neurological developments in neonates.

Child Development ; Erythropoietin ; therapeutic use ; Female ; Humans ; Hypoxia-Ischemia, Brain ; drug therapy ; psychology ; Infant Behavior ; Infant, Newborn ; Male

OBJECTIVETo evaluate the clinical efficacy and safety of angong niuhuang pill (ANP) as an adjuvant treatment on moderate or severe neonatal hypoxic-ischemic encephalopathy (NHIE).

METHODSThirty-nine neonates with NHIE in the control group were treated with conventional treatment, and 58 in the treated group were administered orally ANP additionally, and relative indexes were observed.

RESULTSThe improvement of aspects such as recovery of consciousness, muscular tension, and primitive reflex and disappearance of convulsion, in the treated group was better than that in the control group (P < 0.01).

CONCLUSIONANP as an adjuvant treatment has a definite effect on NHIE, it can promote the recovery of patients, decrease the occurrence of sequelae and with high safety, therefore, is a drug feasible for clinical application.

Asphyxia Neonatorum ; complications ; drug therapy ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypoxia-Ischemia, Brain ; drug therapy ; etiology ; Infant, Newborn ; Male ; Phytotherapy

We have previously shown that cycloheximide significantly inhibited apoptosis, and reduced ensuing cerebral infarction in a newborn rat model of cerebral hypoxiaischemia. This study was performed to determine the therapeutic window for cycloheximide therapy. Seven day-old newborn rat pups were subjected to 100 min of 8% oxygen following a unilateral carotid artery ligation, and cycloheximide was given at 0, 6, 12 and 24 hr after hypoxia-ischemia (HI). Apoptosis or necrosis was identified by performing flow cytometry with a combination of fluorescinated annexin V and propidium iodide, and the extent of cerebral infarction was evaluated with triphenyl tetrazolium chloride (TTC) at 48 hr and 72 hr after HI, respectively. With cycloheximide treatment at 0 hr after HI, both apoptotic and necrotic cells by flow cytometry were significantly reduced, only necrotic cells were significantly reduced at 6 and 12 hr, and no protective effect was seen if administration was delayed until 24 hr after HI compared to the HI control group. Infarct volume, measured by TTC, was significantly reduced by 92% and 61% when cycloheximide was given at 0 or 6 hr after HI respectively; however, there was an insignificant trend in infarct reduction if cycloheximide was administered 12 hr after HI, and no protective effect was observed when administration was delayed until 24 hr after HI. In summary, cycloheximide was neuroprotective when given within 6 hr after HI in the developing newborn rat brain.
Rats, Sprague-Dawley ; Rats ; Protein Synthesis Inhibitors/therapeutic use ; Oxygen/metabolism ; Neuroprotective Agents/*therapeutic use ; Necrosis ; Hypoxia-Ischemia, Brain/*drug therapy ; Hypoxia, Brain ; Humans ; Flow Cytometry ; Cycloheximide/*therapeutic use ; Brain Ischemia ; Apoptosis ; Animals, Newborn ; Animals