Comparative study of the distribution and expression of Neuroglobin and Hypoxia-inducible factor-1α in the adult and young Yak Brain / Estudo comparativo da distribuição e expressão de neuroglobina e fator-1α indutível por hipóxia em adultos e jovens iaques (Yak Brain)

Background: The brain is an organ that serves as the center of the nervous system in all vertebrate and most invertebrate animals. Aim: The study examined the expression of Neuroglobin (Ngb) and Hypoxia-inducible factor-1α (Hif-1α) in adult and young yak brain tissues, and provided researchers with meaningful insight into the anatomy, physiology, and biochemistry of this mammal. Method: The study employed immunohistochemistry (IHC), quantitative real-time PCR (qRT-PCR), and Western blot (WB) to obtain the results. Results: Ngb and Hif-1α were significantly (P<0.05) expressed in the cerebellar cortex, piriform lobe, medulla, and corpus callosum of the adult yak while in the young yak brain tissues, the protein expressions were significantly found in the white matter of the cerebellum, pineal gland, corpus callosum, and cerebellar cortex. The Ngb and Hif-1α expression showed similarities and differences. This may have resulted from similar animal species, source of nutrition, age factors, brain size, emotional activities, and communication. The findings documented that Ngb and Hif-1α are commonly expressed in various adult and young yak brain tissues. Multiple roles in the brain tissues of the adult and young yaks are involved in the expression and distribution and are proposed to play a significant role in the adaptation of the yak to the high altitude environment. Conclusion: This study provides meaningful data to understand the adaptive mechanism to hypoxia and recommended researchers to expand on the adaptive mechanism and brain tissues that are not recorded.

Contexto: O cérebro é um órgão que funciona como o centro do sistema nervoso em todos os animais vertebrados e na maioria dos invertebrados. Objetivo: O estudo examinou a expressão de neuroglobina (Ngb) e fator-1α indutível por hipóxia (Hif-1α) em tecidos cerebrais de iaques adultos e jovens e forneceu aos pesquisadores uma visão significativa da anatomia, fisiologia e bioquímica desse mamífero. Método: O estudo utilizou imuno-histoquímica (IHC), PCR quantitativo em tempo real (qRT-PCR) e western blot (WB) para a obtenção dos resultados. Resultados: Ngb e Hif-1α foram significativamente (P < 0,05) expressos no córtex cerebelar, lobo piriforme, medula e corpo caloso do iaque adulto, enquanto nos tecidos cerebrais do iaque jovem as expressões proteicas foram encontradas significativamente na substância branca do cerebelo, glândula pineal, corpo caloso e córtex cerebelar. A expressão de Ngb e Hif-1α apresentou semelhanças e diferenças. Isso pode ter resultado de espécies animais semelhantes, fonte de nutrição, fatores de idade, tamanho do cérebro, atividades emocionais e comunicação. Os resultados documentaram que o Ngb e o Hif-1α são comumente expressos em vários tecidos cerebrais de iaques adultos e jovens. Múltiplos papéis nos tecidos cerebrais de iaques adultos e jovens estão envolvidos na expressão e distribuição e são propostos para desempenhar um papel significativo na adaptação do iaque ao ambiente de alta altitude. Conclusão: Este estudo fornece dados significativos para compreender o mecanismo adaptativo à hipóxia e recomendou que os pesquisadores expandissem o mecanismo adaptativo e os tecidos cerebrais que não foram registrados.

Comparative study of the distribution and expression of Neuroglobin and Hypoxia-inducible factor-1 in the adult and young Yak Brain

Abstract Background The brain is an organ that serves as the center of the nervous system in all vertebrate and most invertebrate animals. Aim The study examined the expression of Neuroglobin (Ngb) and Hypoxia-inducible factor-1 (Hif-1) in adult and young yak brain tissues, and provided researchers with meaningful insight into the anatomy, physiology, and biochemistry of this mammal. Method The study employed immunohistochemistry (IHC), quantitative real-time PCR (qRT-PCR), and Western blot (WB) to obtain the results. Results Ngb and Hif-1 were significantly (P 0.05) expressed in the cerebellar cortex, piriform lobe, medulla, and corpus callosum of the adult yak while in the young yak brain tissues, the protein expressions were significantly found in the white matter of the cerebellum, pineal gland, corpus callosum, and cerebellar cortex. The Ngb and Hif-1 expression showed similarities and differences. This may have resulted from similar animal species, source of nutrition, age factors, brain size, emotional activities, and communication. The findings documented that Ngb and Hif-1 are commonly expressed in various adult and young yak brain tissues. Multiple roles in the brain tissues of the adult and young yaks are involved in the expression and distribution and are proposed to play a significant role in the adaptation of the yak to the high altitude environment. Conclusion This study provides meaningful data to understand the adaptive mechanism to hypoxia and recommended researchers to expand on the adaptive mechanism and brain tissues that are not recorded.

Resumo Contexto O cérebro é um órgão que funciona como o centro do sistema nervoso em todos os animais vertebrados e na maioria dos invertebrados. Objetivo O estudo examinou a expressão de neuroglobina (Ngb) e fator-1 indutível por hipóxia (Hif-1) em tecidos cerebrais de iaques adultos e jovens e forneceu aos pesquisadores uma visão significativa da anatomia, fisiologia e bioquímica desse mamífero. Método O estudo utilizou imuno-histoquímica (IHC), PCR quantitativo em tempo real (qRT-PCR) e western blot (WB) para a obtenção dos resultados. Resultados Ngb e Hif-1 foram significativamente (P 0,05) expressos no córtex cerebelar, lobo piriforme, medula e corpo caloso do iaque adulto, enquanto nos tecidos cerebrais do iaque jovem as expressões proteicas foram encontradas significativamente na substância branca do cerebelo, glândula pineal, corpo caloso e córtex cerebelar. A expressão de Ngb e Hif-1 apresentou semelhanças e diferenças. Isso pode ter resultado de espécies animais semelhantes, fonte de nutrição, fatores de idade, tamanho do cérebro, atividades emocionais e comunicação. Os resultados documentaram que o Ngb e o Hif-1 são comumente expressos em vários tecidos cerebrais de iaques adultos e jovens. Múltiplos papéis nos tecidos cerebrais de iaques adultos e jovens estão envolvidos na expressão e distribuição e são propostos para desempenhar um papel significativo na adaptação do iaque ao ambiente de alta altitude. Conclusão Este estudo fornece dados significativos para compreender o mecanismo adaptativo à hipóxia e recomendou que os pesquisadores expandissem o mecanismo adaptativo e os tecidos cerebrais que não foram registrados.

Comparative study of the distribution and expression of Neuroglobin and Hypoxia-inducible factor-1α in the adult and young Yak Brain / Estudo comparativo da distribuição e expressão de neuroglobina e fator-1α indutível por hipóxia em adultos e jovens iaques (Yak Brain)

Abstract Background The brain is an organ that serves as the center of the nervous system in all vertebrate and most invertebrate animals. Aim The study examined the expression of Neuroglobin (Ngb) and Hypoxia-inducible factor-1α (Hif-1α) in adult and young yak brain tissues, and provided researchers with meaningful insight into the anatomy, physiology, and biochemistry of this mammal. Method The study employed immunohistochemistry (IHC), quantitative real-time PCR (qRT-PCR), and Western blot (WB) to obtain the results. Results Ngb and Hif-1α were significantly (P<0.05) expressed in the cerebellar cortex, piriform lobe, medulla, and corpus callosum of the adult yak while in the young yak brain tissues, the protein expressions were significantly found in the white matter of the cerebellum, pineal gland, corpus callosum, and cerebellar cortex. The Ngb and Hif-1α expression showed similarities and differences. This may have resulted from similar animal species, source of nutrition, age factors, brain size, emotional activities, and communication. The findings documented that Ngb and Hif-1α are commonly expressed in various adult and young yak brain tissues. Multiple roles in the brain tissues of the adult and young yaks are involved in the expression and distribution and are proposed to play a significant role in the adaptation of the yak to the high altitude environment. Conclusion This study provides meaningful data to understand the adaptive mechanism to hypoxia and recommended researchers to expand on the adaptive mechanism and brain tissues that are not recorded.

Resumo Contexto O cérebro é um órgão que funciona como o centro do sistema nervoso em todos os animais vertebrados e na maioria dos invertebrados. Objetivo O estudo examinou a expressão de neuroglobina (Ngb) e fator-1α indutível por hipóxia (Hif-1α) em tecidos cerebrais de iaques adultos e jovens e forneceu aos pesquisadores uma visão significativa da anatomia, fisiologia e bioquímica desse mamífero. Método O estudo utilizou imuno-histoquímica (IHC), PCR quantitativo em tempo real (qRT-PCR) e western blot (WB) para a obtenção dos resultados. Resultados Ngb e Hif-1α foram significativamente (P < 0,05) expressos no córtex cerebelar, lobo piriforme, medula e corpo caloso do iaque adulto, enquanto nos tecidos cerebrais do iaque jovem as expressões proteicas foram encontradas significativamente na substância branca do cerebelo, glândula pineal, corpo caloso e córtex cerebelar. A expressão de Ngb e Hif-1α apresentou semelhanças e diferenças. Isso pode ter resultado de espécies animais semelhantes, fonte de nutrição, fatores de idade, tamanho do cérebro, atividades emocionais e comunicação. Os resultados documentaram que o Ngb e o Hif-1α são comumente expressos em vários tecidos cerebrais de iaques adultos e jovens. Múltiplos papéis nos tecidos cerebrais de iaques adultos e jovens estão envolvidos na expressão e distribuição e são propostos para desempenhar um papel significativo na adaptação do iaque ao ambiente de alta altitude. Conclusão Este estudo fornece dados significativos para compreender o mecanismo adaptativo à hipóxia e recomendou que os pesquisadores expandissem o mecanismo adaptativo e os tecidos cerebrais que não foram registrados.

Effect of acupuncture on learning and memory ability and hippocampal Ngb expression in rats with vascular cognitive impairment / 国际中医中药杂志

Objective:To discuss the effect of acupuncture on learning and memory ability and the expression of neuroglobin (Ngb) of the hippocampus of vascular cognitive impaired (VCI) rats.Methods:VCI rats model was established by repeated ligation of the carotid arteries of the two sides to induce ischemia-reperfusion injury. The successful modeled rats were divided into acupuncture group and a model group according to the random number table method, with 12 rats in each group, and 12 rats were selected as the normal group. The rats in the acupuncture group received acupuncture at "Baihui", "Fengfu" and "Yongquan", 10 times as a course of treatment, a total of 3 courses, and stop acupuncture treatment for 2 days during the courses of treatment. Neurobehavioral scores were measured on the 3rd day after the intervention. Morris water maze was conducted to detect the learning and memory ability of the rats in each group, and Western blot was used to detect the expression of Ngb in the hippocampus of the rats.Results:Compared with the model group, the behavioral score of the acupuncture group decreased after intervention ( P<0.05), the escape latency was significantly shortened ( P<0.05), the number of crossing platforms was increased ( P<0.05), and the hippocampal Ngb (1.18±0.06 vs.0.98±0.04) was significantly increased ( P<0.05). Conclusion:Acupuncture at "Baihui", "Fengfu" and "Yongquan" could promote the expression of Ngb in the hippocampus of rats and improve the learning and memory ability of VCI rats.

The effect of rehabilitation training on the expression of neuroglobin in the peri-infarct cortex and its mechanism / 中华物理医学与康复杂志

Objective:To study the effect of rehabilitation training on the expression of neuroglobin (Ngb), oxidative stress and axon regeneration in the cortex and explore possible mechanisms of functional recovery after cerebral infarction.Methods:Thirty-six male Sprague-Dawley rats were randomly divided into a sham operation group, a model group and a rehabilitation group. Cerebral infarction was modelled in the model and rehabilitation groups using Longa′s middle cerebral artery occlusion (MCAO) technique. The sham operation group received the same procedure except that no thread was inserted to block the middle cerebral artery. The rats in the rehabilitation group began treadmill training 24h after the operation, while the other two groups were left on the treadmill without training. On the 3rd, 7th and 14th days after the operation, all of the rats′ neurological functioning was assessed using modified neurological severity scores (mNSSs). After the last mNSS test, all of the rats were sacrificed and peri-infarct brain tissue was resected to detect the expression of Ngb and oxidative stress indicators including superoxide dismutase (SOD), nitric oxide and malondialdehyde (MDA), as well as neurofilament-200 (NF-200) indicating axon regeneration.Results:On the 3rd day after the surgery there was no significant difference between the average mNSS scores of the rehabilitation and model groups. On the 7th and 14th day the average mNSS score of the rehabilitation group was significantly better than that of the model group. The average expression of Ngb in the model group was significantly higher than in the sham operation group. After the intervention, the average expression of SOD in the rehabilitation group was significantly higher than in the model group, while NO and MDA expression were significantly lower. After the intervention the average expression of NF-200 in the rehabilitation group was also significantly higher than in the model group.Conclusions:Rehabilitation training benefits the recovery of neurological function after cerebral infarction, at least in rats. The mechanism may be related to the upregulation of Ngb, alleviation of oxidative stress and enhancement of axonal regeneration in the peri-infarct cortex.

Mechanism of dexmedetomidine alleviating sevoflurane-induced neurotoxicity in neonatal rats: relationship with Ngb / 中华麻醉学杂志

Objective:To evaluate the mechanism of dexmedetomidine alleviating sevoflurane-induced neurotoxicity and the relationship with neuroglobin (Ngb) in neonatal rats.Methods:Eighty-four healthy male Sprague-Dawley rats, aged 7 days, weighing 12-16 g, were divided into 4 groups ( n=21 each) using a random number table method: control group (group C), sevoflurane group (group S), sevoflurane plus dexmedetomidine group (group SD), and sevoflurane plus dexmedetomidine and yohimbine group (group SDY). Sevoflurane anesthesia method: anesthesia was induced by inhaling 8% sevoflurane and then maintained by inhaling 3% sevoflurane for 4 h. At the end of anesthesia, dexmedetomidine 50 μg/kg was intraperitoneally injected in group SD, and 0.5 mg/kg yohimbine (α 2 adrenergic receptor antagonist) was intraperitoneally injected immediately after the end of intraperitoneal injection of dexmedetomidine 50 μg/kg in group SDY.Six rats were randomly selected from each group and sacrificed at 24 h after the end of anesthesia.The brain tissues were removed for determination of histopathologic changes in hippocampal CA1 region (using HE staining), the expression of Ngb in the area of the hippocampus (by immunofluorescence staining) and the expression of cytochrome C (Cyt c) and caspase-3 (by Western blot). The other rats were fed until 28 days old, and Morris water maze test was performed. Results:Compared with group C, the expression of Ngb, Cyt c and caspase-3 was significantly up-regulated ( P<0.05), histopathologic changes in hippocampal CA1 region were accentuated, and Ngb fluorescence intensity was increased in group S. Compared with group S, the expression of Ngb was significantly up-regulated, expression of Cyt c and caspase-3 was down-regulated, the escape latency was shortened ( P<0.05), histopathologic changes in hippocampal CA1 region were reduced, and Ngb fluorescence intensity was increased in group SD.Compared with group SD, the expression of Ngb was significantly down-regulated, expression of Cyt c and caspase-3 was up-regulated, the escape latency was prolonged ( P<0.05), histopathologic changes in hippocampal CA1 region were accentuated, and Ngb fluorescence intensity was decreased in group SDY. Conclusion:The mechanism by which dexmedetomidine reduces sevoflurane-induced neurotoxicity may be related to activating α 2 adrenergic receptors and up-regulating the expression of Ngb in neonatal rats.

Relationship between ubiquitin C-terminal hydrolase-L1, neuroglobinB and neurological prognosis in patients with coma after cardiopulmonary resuscitation / 中国综合临床

Objective:To investigate the relationship between serum ubiquitin C-terminal hydrolase-L1 (UCH-L1) and neuroglobin levels and the prognosis of neurological function in coma patients after cardiopulmonary resuscitation, and to analyze their value in predicting the prognosis of patients.Methods:From February 2018 to June 2020, 45 comatose patients admitted to the Chengdu Third People′s Hospital of Sichuan Province after successful cardiopulmonary resuscitation were prospectively selected as the coma group, and 62 patients admitted to the emergency intensive care unit during the same period after successful cardiopulmonary resuscitation with consciousness recovered within 24 hours were selected as the control group.Serum UCH-L1 and neuroglobin levels were detected within 24 hours after admission.Glasgow coma Scale (GCS)and cerebral performance category(CPC)were used to evaluate coma severity and neurological prognosis.Spearman rank correlation analyzed the correlation between UCH-L1 and neuroglobin levels and GCS and CPC scores.Logistic regression analyzed the factors affecting the prognosis of neurological function in coma patients after cardiopulmonary resuscitation.Receiver operating characteristic curve (ROC) was used to analyze the value of UCH-L1 and neuroglobin in predicting the prognosis of neurological function in coma patients after successful cardiopulmonary resuscitation.Results:In coma group, serum UCH-L1((0.63±0.21) μg/L) and the concentration of neuroglobin ((89.34±21.35) mg/L) was higher than that in the control group ((0.27±0.08) μg/L, (32.13±9.21) mg/L), the difference was statistically significant( t=12.338, 18.846; all P<0.001). The levels of UCH-L1 and neuroglobin in mild, medium and severe coma groups were increased in turn, and the differences between the groups were statistically significant( F=86.430, 26.958; all P<0.001). The serum levels of UCH-L1((0.72±0.06) μg/L)and neuroglobin ((100.35±5.79) mg/L)in the group with poor neurological prognosis were higher than those in the group with good neurological prognosis((0.52±0.08) μg/L, (75.58±6.91) mg/L), and the differences between the groups were statistically significant( t=9.585, 13.086; all P<0.001). UCH-L1 and neuroglobin were negatively correlated with GCS score(rs=-0.685, -0.669; all P<0.001), and positively correlated with CPC score (rs=0.688, 0.670; all P<0.001). Multivariate Logistic regression analysis showed that low GCS score( OR=0.552, 95% CI: 0.392-0.776, P<0.001), high UCH-L1 ( OR=1.881, 95% CI: 1.276-2.773, P<0.001)and neuroglobin( OR=1.677, 95% CI: 1.206-2.331, P=0.001)were independently associated with poor neurological outcomes in coma patients after cardiopulmonary resuscitation .The AUC of combining UCH-L1 and neuroglobin in predicting poor neurological outcomes in coma patients after cardiopulmonary resuscitation was 0.954, which was higher than that of UCH-L1 and neuroglobin alone (0.821, 0.790) ( Z=2.351, 2.649; all P<0.05). Conclusion:After successful cardiopulmonary resuscitation, the levels of UCH-L1 and neuroglobin in coma patients are increased.High levels of UCH-L1 and neuroglobin are associated with coma severity and neurological dysfunction, which can be used as a potential biological indicator for prognosis evaluation of neurological function.

Effects of Hemin on cognitive function and the expression of neuroglobin in rats with vascular de-mentia / 中华行为医学与脑科学杂志

Objective To observe the influence of Hemin on both the expression of neuroglobin (Ngb) and Tau protein phosphorylation in rats with vascular dementia (VD),and explore the effect and mechanism of Ngb in VD. Methods VD model was established in rats with a permanent bilateral common carotid arteries occlusion(2-VO). Fifty four healthy male SD rats were randomly divided into 3 groups:sham operation group,VD group and Hemin group. According to ischemic time,the animals in each group were randomly allocated into 1 week subgroup,4 weeks subgroup and 8 weeks subgroup(n=6 in each subgroup). The learning and memory ability of rats were evaluated by Morris water maze test. The brain histopathological changes were observed by HE staining and the expression of Ngb and phosphorylated Tau ( p-Tau) was de-termined by Western blot. Results Compared with the sham operation group,after 2-VO the escape latency extended (all P<0. 05) and cross-platform performance reduced in rats in both VD group and Hemin group (all P<0. 05). The learning and memory ability of rats in Hemin group was better than that in VD group at the corresponding time point:the increase in the number of crossing platforms(1 week:Hemin group (7. 00± 0. 89) times vs VD group (5. 50±1. 22) times,t=2. 42,P<0. 05;4 weeks:Hemin group (5. 33±0. 52) times vs VD group (3. 50±1. 04) times,t=3. 84,P<0. 01;8 weeks:Hemin group (6. 50±0. 55) times vs VD group (4. 50±1. 05) times,t=4. 14,P<0. 01). The expression of Ngb and phosphorylated Tau (p-Tau) increased in VD group and Hemin group (P<0. 01). Compared with VD group,the expression of Ngb in Hemin group was increased(1 week:Hemin group (3. 45±0. 23) vs VD group (2. 56±0. 08),t=5. 67,P<0. 01;4 weeks:Hemin group (3. 08±0. 13) vs VD group (2. 28±0. 08),t=7. 96,P<0. 01;8 weeks:Hemin group (2. 82± 0. 12) vs VD group (2. 05±0. 10),t=7. 28,P<0. 01),while p-Tau was decreased (1 week:Hemin group (1. 57±0. 12) vs VD group (2. 40±0. 15),t=7. 62,P<0. 01;4 weeks:Hemin group ( 2. 14±0. 21) vs VD group (3. 18±0. 14),t=8. 23,P<0. 01;8 weeks:Hemin group (1. 83±0. 13) vs VD group (2. 79±0. 19), t=4. 91,P<0. 01). In VD group and Hemin group,the learning and memory ability exhibited a negative cor-relation with Ngb (r=-0. 892,P<0. 01),whereas a positive correlation with p-Tau (r=0. 932,P<0. 01). Conclusion Hemin induces high expression of Ngb and inhibits Tau phosphorylation in VD rats. Ngb may play a neuroprotective role in the development of VD by regulating Tau phosphorylation.

The protective effects of ulinastatin on brain tissue after CPR in swine / 中华急诊医学杂志

Objective To study the protective effects of ulinastatin on brain tissue from pathomorphism and the changes of serum IL-6,Ngb (neuroglobin of brain),S100B protein and superoxide dismutase (SOD) after cardiopulmonary resuscitation (CPR) in swine.Methods The CPR model of swine was made by a programmed electric stimulation given to myocardium to produce a ventricular fibrillation and then a CPR was given.A flock of 16 male healthy Beijing landraces pigs of 3-4 months old were divided in random number method into control group (n =6) and ulinastatin group (n =6),because there were 12 swine survived from ventricular fibrillation.The t-test was used for the statistical analysis.ELISA was used to detect the changes in levels of those biomarkers in serum at each interval as well as the pathomorphological changes in brain tissues were observed under the light microscope after HE staining.Results (1) Before ventricular fibrillation,there were no distinct differences in levels of various biomarkers in porcine serum between two groups.After ventricular fibrillation,serum IL-6,S100B protein and Ngb levels in both groups gradually increased as time elapsed,and the levels of those biomarkers in the control were significantly higher than those in the ulinastatin group (P ＜ 0.05).Serum SOD in both groups gradually reduced,and more distinct decline of biomarkers was found in the ulinastatin group (P ＜ 0.05).(2) HE staining showed the porcine brain tissues in control group had significant ischemia,degeneration and necrosis and the degree of those pathological changes in the ulinastatin group were significantly moderated.Conclusion The immediate administration of ulinastatin as CPR initiated can alleviate porcine brain tissue damage after ischemia/reperfusion (I/R).This showed a protective effect of ulinastatin against I/R injury on porcine brain.

The role of neuroglobin in oxygen-glucose deprivation and reoxygenation-induced mitochondrial ;depolarization and reactive oxygen species production in SH-SY5Y cells / 中华内科杂志

Objective To investigate the role of neuroglobin ( NGB) in oxygen-glucose deprivation and reoxygenation ( OGD/R ) induced mitochondrial depolarization and reactive oxygen species ( ROS ) production in a human neuroblastoma cell line (SH-SY5Y).Methods SH-SY5Y cells were transfected with lentivirus to establish a stable cell line of NGB knockdown ( KD).After treated with OGD/R, cells were collected at different time points to analyze NGB mRNA and protein levels.Furthermore, cells were stained with JC-1 and DCFH-DA to evaluate mitochondrial depolarization and ROS production by inverted fluorescence microscope.Also, to determine the neurotoxicity , we measured the lactate dehydrogenase ( LDH) level in the cell culture medium.Results After the treatment of OGD/R, the NGB mRNA and protein started to elevate and peak at 4 h and 8 h (2.04 ±0.35 fold,1.69 ±0.18 fold).Compared with the vector group , NGB KD group had much more mitochondrial depolarization [ JC-1 red/green ( 1.10 ±0.10 ) vs (1.46 ±0.11),P<0.05] and ROS production [DCFH-DA fluorescence (36.30 ±5.32) vs (16.26 ± 2.97),P<0.05].Furthermore, NGB KD groups had a higher level of LDH release [(63.42 ±6.14)%vs (49.65 ±5.09 )%, P <0.05 ].Conclusions NGB plays an important role in the homeostasis of mitochondria.Knockdown of NGB results in increased mitochondrial depolarization , ROS production and neurotoxicity under hypoxia circumstances.

Protective effects of neuroglobin in arsenic-induced nervous system injury / 中华地方病学杂志

Neuroglobin (Ngb) is a globin with high affinity to oxygen and is specifically expressed in the brain of vertebrates.Many studies show that Ngb's expressions change and it may have protective effects in nervous system injury,such as ischemia/hypoxia.Mechanisms of the Ngb's protective role include free radicals scavenging,anti-oxidative stress,as well as anti-apoptosis.Ngb can also play a transduction signal to increase downstream phosphatidylinositol-3-kinase/protein serine threonine kinase (PI3K/AKT) signal activities.Arsenic exposure can lead to oxidative stress and apoptosis,Ngb may play a protective role in nervous system injury.In this article,we summarized expression changes and protective role of neuroglobin in nervous system injuries,and out looked the protective role of neuroglobin in endemic arsenic poisoning induced nervous system injury.

Protein content of neuroglobin in rat brain with septic shock / 临床麻醉学杂志

Objective To observe the expression of neuroglobin in brain of rat with septic shock,and study the protective effect of neuroglobin.Methods Sixty male SD rats were randomly di-vided into two groups,30 rats in each.The rats in group M were injected LPS 10 mg/kg through tail vein to establish septic shock model,the success standard of septic shock was systolic blood pressure dropping to 60% of the basic values;the control group were given equal volume of NS.The levels of rat TNF-α,IL-6 and NGB in serum at the time points of before LPS injection (T0 ),1 h (T1 ),2 h (T2 ),4 h (T3 )and 6 h(T4 )after LPS injection were measured using ELISA kits.NGB and caspase-3 protein expression in brain were determined using Western blotting.Results Compared with group C and T0 ,the TNF-α,IL-6 concentrations in serum and caspase-3 in brain were increased significantly at T1-T4 (P <0.05),NGB protein content in serum and brain at T3 and T4 were increased significantly (P <0.05).Conclusion Cell apoptosis and inflammatory response are discovered in the brain during septic shock.NGB is over-expressed in brain of rats with septic shock,which may play a role of pro-tective function.

Relationship of serum neuroglobin and neuron-specific enolase level in preterm infants with brain damage / 国际儿科学杂志

Objective To study the relationships of serum neuroglobin and neuron-specific enolase level with periventricular hemorrhage-intraventricular hemorrhage ( PVH-IVH) and periventricular leucumalacia ( PVL) in preterm infants. Methods There were 241 cases of preterm infants whose gestational age was less than 34 weeks and were admitted in NICU of Guangzhou Women and Children′s Medical Center, Guangzhou Huadu District Matermal and Child Health Hospital and Dongguan Taiping Hospital from Jan. 2010 to May. 2013, enrolled in the study. The serum level of neuroglobin and neuron-specific enolase were detected within 12 hours and on the 3 d, 7 d, 14 d after birth. Cranial ultrasound was preformed 2~3 d, 1week, 2weeks, 3weeks, and 4 weeks after birth. They also received Cranial MRI examination before discharge or when the correct gestational age reached 40 weeks. All 241 cases were divided into 3 groups ( no brain damage group, PVH-IVH group and PVL group) according to the result of cranial US and MRI. The differences of the serum levels of neuroglobin and neuron-specific enolase among each groups were compared. Results The results of cranial ultrasound and /or MRI showed: 162 cases had no brain damage ( in no brain damage group) , 50 cases had PVH-IVH ( in PVH-IVH group) , and 20 cases had PVL, 9 cases had PVL and PVH-IVH ( both in PVL group) . Within 12 h and 3 d after birth, the serum levels of neuroglobin in PVL group and PVH-IVH group was significantly higher than those in no brain damage group (P0. 05 ) , and there were still significantly higher than those in no brain damage group and PVH-IVH group (all P0. 05). On 7 d and 14 d after birth, the serum levels of neuron-specific enolase in PVL group were no significant difference compared with PVH-IVH group and no brain damage group (all P>0. 05). Conclusion The increased serum levels of neuroglobin and neuron-specific enolase in preterm infants within 12 h and 3 d after birth would have certain clinical significance for judging whether early brain damage and PVL would happen.

Overexpression of neuroglobin attenuates Aβ-induced apoptosis in brains of double transgenic AD mice / 中国病理生理杂志

AIM:To observe the effects of neuroglobin ( NGB) overexpression on the apoptosis induced by Aβin the brains of double transgenic AD ( APPswe/PS1dE9) mice and to explore its potential mechanisms .METHODS:Twenty-four 13-month-old double transgenic AD mice were randomly divided into 3 groups:intracerebroventricular injection with normal saline (NS) group, intracerebroventricular injection with pcDNA3.1 and NS group, and intracerebroventricular injection with pcDNA3.1 and pNGB group.Immunohistochemistry was used to detect the expression of Aβ1-42 in the brains. TUNEL staining was used for analyzing the apoptosis , and the protein levels of cleaved caspase-3, caspase-9, PI3K, Akt and p-Akt were determined by Western blot .RESULTS: After intracerebroventricular injection with pNGB , the areas of Aβ1-42 in the hippocampus and cortex were decreased compared with NS group and pcDNA 3.1+NS group (P<0.01).The TUNEL-positive staining cells in the pNGB group were less than those in NS group and pcDNA 3.1 group ( P<0.01 ) . NGB overexpression attenuated the protein levels of cleaved caspase-3 and caspase-9 (P<0.01), but induced the produc-tion of PI3K and p-Akt (P<0.01).CONCLUSION:Overexpression of pNGB significantly inhibits the generation of Aβand attenuates the apoptosis induced by Aβ, indicating that NGB overexpression activates PI 3K/Akt pathway and inhibits the production of cleaved caspase-3 and caspase-9, which were tightly related with apoptosis .

Expression of hypoxia-inducible factor 1 and neuroglobin in piglet cortex during deep hypothermic circulatory arrest / 中国病理生理杂志

AIM:To observe the expression of hypoxia-inducible factor 1 (HIF-1) and neuroglobin (NGB) in piglet cortex during deep hypothermic circulatory arrest.METHODS:Wuzhishan piglets were randomly assigned to car-diopulmonary bypass group ( CPB group) , 40 min of circulatory arrest ( CA) at 18 ℃ without cerebral perfusion ( DHCA group) or with selective antegrade cerebral perfusion ( SACP group) .After 180 min of reperfusion, cortical tissue was har-vested for determining HIF-1αand NGB expression by HE staining, Western blot and real-time PCR.RESULTS:Severer cerebral injury was observed in DHCA group than that in SACP group.After 180 min of reperfusion, HIF-1αprotein and mRNA levels were significantly higher in DHCA group than those in CPB group (P<0.05).Accordingly, SACP animal had higher levels of HIF-1αprotein and mRNA than those in DHCA group (P<0.05).Simultaneously, higher NGB pro-tein and mRNA levels were found in DHCA group than those in CPB group after 180 min of reperfusion ( P<0.05) .The SACP animal had higher levels of NGB protein and mRNA than those in DHCA group (P<0.05).CONCLUSION:Up-regulation of HIF-1 and NGB are involved in the mechanism against cerebral injury resulting from DHCA in the cortex and possibly a part of cerebral protective effect of SACP.

Effects of sodium arsenite on cell apoptosis in rat cerebellar granule and nerve globin mRNA expression / 中华地方病学杂志

Objective To observe the effects of sodium arsenite (NaAsO2) on apoptosis in primary cultured rat cerebellar granule cell and nerve globin mRNA expression.Methods Twenty 7-8 days postnatal Wistar rats were selected and decapitated cerebellar cortex, granular cell suspension was prepared and 4 × 109/m2 cells were seeded in the culture flask.Cells were exposed to different concentrations NaAsO2 [0 (control), 2, 5 and 10 μmol/L] for 24 h and intracellular reactive oxygen species (ROS) content and apoptosis were determined by laser confocal,while real-time quantitative PCR was used to detect mRNA expression in cultured cells after 24 and 48 h cell culture.Results Fluorescence intensity of ROS of 0, 2, 5, 10 μmol/L fluoride groups were 421.32 ± 53.52,1 082.09 ± 321.58, 2 223.02 ± 1 011.37 and 1 366.87 ± 102.14, the difference was statistically significant (F =5.873,P ＜ 0.05);Apoptosis rates were (8.91 ± 5.63)％, (26.46 ± 1.77)％, (43.65 ± 14.45)％ and (56.04 ± 6.95)％, the difference was statistically significant (F =18.369, P ＜ 0.05);24 h nerve globin mRNA expression were 1.00 ± 0.06,0.77 ± 0.22, 0.43 ± 0.18 and 0.42 ± 0.14, the difference was statistically significant (F =18.235, P ＜ 0.05);48 h nerve globin mRNA expression were 1.00 ± 0.01, 0.89 ± 0.09, 0.60 ± 0.16 and 0.08 ± 0.02, the difference was statistically significant (F =80.843, P ＜ 0.05).Conclusion With increased NaAsO2 concentration, ROS and apoptosis levels are increased, while nerve globin mRNA expression is significantly reduced, suggesting that after the body is stimulated, protective proteins might be consumed, further suggesting nerve globin expression might be involved in nerve damage caused by arsenic.

Roles and their mechanisms of neuroglobin in cerebral ischemia / 国际脑血管病杂志

Neuroglobin (Ngb) is an oxygen-carrying globin that specifically expresses in brain tissue.It is involved in energy metabolism,mitochondrial function,as well as cell survival and proliferation of signaling pathway regulation.Under physiological conditions,Ngb presents as a form of ferrous deoxy hexacoordination,which has stronger oxygen affinity.During ischemia and hypoxia,the expression of Ngb is upregulated in brain tissue and interacts through a variety of proteins of its downstream,and plays a protective role for the damaged brain tissue.

Prevention and treatment of AD by over-expression of neuroglobin via ac-tivating PI3K/Akt signaling pathway / 中国病理生理杂志

AIM: To study the neuroprotective roles of neuroglobin (NGB) over-expression in the SH-SY5Y cells transfected with pAPPswe.METHODS:The plasmid pEGFP-NGB was successfully constructed and transfected into the SH-SY5Y cells, which were pretreated with pAPPswe.MTT assay was applied to detect the effect of NGB over-expres-sion on the cell survival rates.JC-1 staining was used to detect the level of mitochondrial transmembrane potential.The cell apoptosis was analyzed by flow cytometry.The effects of NGB over-expression on the protein level of p-Akt, Akt and caspase-3/9 were determined by Western blotting.The generation of Aβ42 in the cells was measured by ELISA.RE-SULTS:The cell survival rate was remarkably increased after transfection with NGB compared with control group and emp-ty plasmid group (P<0.05).The over-expression of NGB significantly inhibited the decrease in mitochondrial membrane potential induced by pAPPswe.The over-expression of NGB inhibited the apoptosis of the cells.Furthermore, over-expres-sion of NGB not only inhibited the expression of caspase-3 and caspase-9, but also induced the production of p-Akt, which was prevented by LY294002, an inhibitor of PI3K/Akt.The generation of Aβ42 was inhibited in the cells with the over-ex-pression of NGB.CONCLUSION: Over-expression of NGB significantly inhibits the SH-SY5Y cell injuries induced by pAPPswe and inhibits the expression of caspase-3/9, which is tightly related with cell apoptosis.Furthermore, the neuro-protective roles of NGB may be via activating PI3K/Akt signaling pathway.

Valproic acid induces neuroglobin protein by CREB and protects N2a cells against H2 O2-induced neurotoxicity / 中国药理学通报

Aim To investigate the effect and mecha-nism of valproic acid on neuroglobin expression, and the neuroprotective role of valproic acid against H2 O2-induced neurotoxicity. Methods Western blot, RT-PCR and luciferase assay were used to detect the pro-tein levels, mRNA levels and promoter activity of mouse and human neuroglobin induced by valproic acid. Luciferase assay was used to investigate the role of transcription factor CREB in the up-regulation of neuroglobin by valproic acid. MTT assay was used to evaluate the effect of valproic acid against H2 O2-in-duced neurotoxicity. Results VPA treatment marked-ly increased the protein levels, mRNA levels and pro-moter activity of Ngb in mouse N2 a cells and human SKNSH cells. CREB specific inhibitor KG501 or CREB dominant negative mutant KCREB attenuated VPA-induced Ngb promoter activity. VPA could pro-tect N2a cells from H2 O2-induced neurotoxicity. Con-clusion CREB mediates VPA-induced Ngb up-regula-tion, which may contribute to the neuroprotective effects of VPA in oxidative stress in neurons.

Progress on the molecular mechanisms of neuroglobin improving neonatal hypoxia-isehemia brain injury / 国际儿科学杂志

Neuroglobin is involved in neuroprotection from damage due to hypoxia or ischemia in vitro and in vivo.Overexpression of neuroglobin ameliorates the recovery from hypoxia-ischemia brain injury in experimental animals.The mechanism is still unknown.Studies have revealed that neuroglobin has a typical globin fold,and despite being hexacoordinated,which binds reversibly O2,CO2,and NO.This article reviews the possible mechanisms involved in neuroprotection of neuroglobin and provide a clue in treatment of hypoxia-ischemia brain injury.