Anti-placental growth factor antibody ameliorates hyperoxia-mediated impairment of lung development in neonatal rats

This study investigates the effect of the overexpression of the placental growth factor (PGF) and hyperoxia on lung development and determines whether anti-PGF antibody ameliorates hyperoxia-mediated impairment of lung development in newborn rats. After exposure to normoxic conditions for seven days, newborn rats subjected to normoxia were intraperitoneally or intratracheally injected with physiological saline, adenovirus-negative control (Ad-NC), or adenovirus-PGF (Ad-PGF) to create the Normoxia, Normoxia+Ad-NC, and Normoxia+Ad-PGF groups, respectively. Newborn rats subjected to hyperoxia were intraperitoneally injected with physiological saline or anti-PGF antibodies to create the Hyperoxia and Hyperoxia+anti-PGF groups, respectively. Our results revealed significant augmentation in the levels of PGF and its receptor Flt-1 in the lung tissues of newborn rats belonging to the Normoxia+Ad-PGF or Hyperoxia groups. PGF overexpression in these groups caused lung injury in newborn rats, while anti-PGF antibody treatment significantly cured the hyperoxia-induced lung injury. Moreover, PGF overexpression significantly increased TNF-α and Il-6 levels in the bronchoalveolar lavage (BAL) fluid of the Normoxia+Ad-PGF and Hyperoxia groups. However, their levels were significantly reduced in the BAL fluid of the Hyperoxia+anti-PGF group. Immunohistochemical analysis revealed that PGF overexpression and hyperoxia treatment significantly increased the expression of the angiogenesis marker, CD34. However, its expression was significantly decreased upon administration of anti-PGF antibodies (compared to the control group under hyperoxia). In conclusion, PGF overexpression impairs lung development in newborn rats while its inhibition using an anti-PGF antibody ameliorates the same. These results provided new insights for the clinical management of bronchopulmonary dysplasia in premature infants.

¿El licopeno puede eliminar los efectos perjudiciales de la hiperoxia en los cerebros inmaduros? / Can lycopene eliminate the harmful effects of hyperoxia in an immature brain?

Objetivos: Al ser un antioxidante, el licopeno protege a las células contra el daño causado por los radicales libres, fortalece los enlaces intercelulares y mejora el metabolismo celular. Este estudio analiza los efectos del licopeno sobre los trastornos neurodegenerativos por hiperoxia en ratas recién nacidas a término. Métodos: Estas ratas se dividieron en cuatro grupos: grupo 1 de referencia con normoxia, grupo 2 con normoxia + licopeno, grupo 3 de referencia con hiperoxia y grupo 4 con hiperoxia + licopeno. Los grupos 1 y 2 se supervisaron en condiciones de aire ambiental, y los grupos 3 y 4 se supervisaron con un nivel de oxígeno > 85 % O2. Los grupos 2 y 4 recibieron inyecciones intraperitoneales de licopeno de 50 mg/kg/día; los otros grupos recibieron inyecciones intraperitoneales de aceite de maíz con el mismo volumen. Las ratas se sacrificaron en el día 11, después de 10 días con hiperoxia. Se extrajeron los cerebros, y se evaluaron los parámetros del sistema oxidativo. Resultados: Se detectaron lesiones cerebrales por hiperoxia en sustancia blanca, regiones corticales y tálamo. Aumentó la cantidad de células apoptóticas y disminuyó la cantidad de células PCNA positivas en los grupos 3 y 4, comparados con el grupo 1. No se observó una mejora significativa en la cantidad de células apoptóticas y células PCNA positivas en los grupos 3 y 4; además, aumentó la apoptosis. Conclusión: Se halló que el licopeno no mostró efectos terapéuticos para el daño cerebral en ratas recién nacidas. Además, se demostró que el licopeno podría causar efectos tóxicos.

Objectives. In addition to protecting cells against free radical harm thanks to its anti-oxidant nature, lycopene strengthens the bonds among cells and improves cell metabolism. This study focuses on analyzing therapeutic effects of lycopene in hyperoxia-induced neurodegenerative disorders in newborn rats. Methods. Term newborn rats were divided into four groups as the normoxia control group (group-1), normoxia+lycopene group (group-2), hyperoxia control group (group-3) and hyperoxia+lycopene group (group-4). Group-1 and group-2 were monitored in room air while the group-3 and group-4 were monitored at > 85% O2. The group-2 and group-4 were injected with lycopene intrapertioneally (i.p. ) at 50mg/kg/day while the other groups were injected with corn oil i.p. at the same volume. The rats we sacrificed on the 11th day following the 10-day hyperoxia. The brains were removed and oxidant system parameters were assessed. Results. Injury resulting from hyperoxia was detected in the white matter, cortical regions, and thalamus of the brains. It was observed that the number of apoptotic cells increased and the number of proliferating cell nuclear antigen (PCNA) positive cells decreased in the groups-3 and 4 compared to the group-1. No significant improvement in the number of apoptotic cells and PCNA positive cells was observed in the groups-3 and 4, and apoptosis increased as well. Conclusion. This study found that lycopene, did not show any therapeutic effects for brain damage treatment in newborn rats. In addition, this study demonstrated that lycopene might lead to toxic effects.

Effects and mechanism of heat shock protein 70 induced by rhubarb on lung development of bronchial lung in newborn rats / 国际儿科学杂志

Objective To establish SD rat model of bronchopulmonary dysplasia(BPD) and investigate the influence and mechanism of heat shock protein 70 (HSP70) on the lung tissue with BPD.Methods One hundred and twenty new born SD rats were randomly equally divided into three groups,including blank-control group,bronchopulmonary dysplasia group,rhubarb intervention group (bronchopulmonary dysplasia rats treated with rhubarb).On the 7th,14th and 21th day,10 rats were euthanasiaed after weighted of each group.Histopathology of pulmonary,hematoxylin and eosin(HE)and the expression of HSP70 in the right lung were observed respectively.Results Compared with the rat of blank-control group,the rat weight of BPD group was significantly decreased at the same time point(P ＜0.01).On the 7th day,the alveolar cavity increased,alveolar wall became thin,and pulmonary interstitial cells increased in the rat of BPD group given oxygen.By 21 days,the normal structure of the alveoli disappeared,the diameter of the alveolar cavity enlarged,the number of alveoli significantly reduced,the alveolar fusion was large,and the alveolar space became thickness,and massive of interstitial cells hyperplasia.While,in the rhubarb intervention group,the lung tissue structure kept integrity,the size of the alveoli uniform.During 14 to 21 days,the integrity of lung structure and the uniform size alveoli still existed,and alveolar septa was thin.In immunohistochemical staining and western blotting experiments,the expression of HSP70 in rhubarb intervention group increased gradually with the increase of age,which was significantly different from control group(P ＜0.01).However,the expression of HSP70 in BPD group had not significant change on the 7th,14th and 21th days(P ＞0.05).Compared with rhubarb intervention group,the expression of HSP70 in the BPD group significantly decreased on the 14th and 21th day(P ＜0.01) respectively.Conclusion Rhubarb could protect the injured lung tissue by increasing the expression of HSP70.

Propofol combined with hypoxia induces cognitive dysfunction in immature rats p38 pathway / 南方医科大学学报

OBJECTIVE@#To investigate the effects of propofol combined with hypoxia on cognitive function of immature rats and the possible role of p38 pathway and tau protein in mediating such effects.@*METHODS@#Ninety 7-day-old (P7) SD rats were randomized for daily intraperitoneal injection of propofol (50 mg/kg) or lipid emulsion (5.0 mL/kg) for 7 consecutive days. After each injection, the rats were placed in a warm box (38 ℃) with an oxygen concentration of 18% (hypoxia), 21% (normal air), or 50% (oxygen) until full recovery of the righting reflex. Another 90 P7 rats were similarly grouped and received intraperitoneal injections of p-p38 blocker (15 mg/kg) 30 min before the same treaments. The phosphorylated tau protein, total tau protein and p-p38 content in the hippocampus were detected using Western blotting. The spatial learning and memory abilities of the rats were evaluated with Morris water maze test.@*RESULTS@#Compared with lipid emulsion, propofol injection resulted in significantly increased levels of p-p38, phosphorylated tau and total tau proteins in rats with subsequent hypoxic or normal air treatment ( < 0.05), but propofol with oxygen and injections of the blocker before propofol did not cause significant changes in the proteins. Without subsequent oxygenation, the rats receiving injections of propofol, with and without prior blocker injection, all showed significantly prolonged latency time and reduced platform-crossing times and third quadrant residence time compared with the corresponding lipid emulsion groups ( < 0.05). With oxygen treatment, the rats in propofoland blocker-treated groups showed no significant difference in the performance in Morris water maze test from the corresponding lipid emulsion group. The results of Morris water maze test differed significantly between blocker-propofol group and propofol groups irrespective of exposures to different oxygen levels ( < 0.05), but not between the lipid emulsion and blocker group pairs with exposures to different oxygen levels.@*CONCLUSIONS@#Propofol combined with hypoxia can affect the expression of tau protein through p38 pathway to impair the cognitive function of immature rats, in which oxygen plays a protective role.

The expression of bone morphogenetic protein 4 and its effects in ischemia-anoxemia induced white matter injury model / 中国小儿急救医学

Objective To investigate the ultrastructural alteration in brain tissues as well as the expression of bone morphogenetic protein(BMP) 4 and its effects on regulating myelination in the process of white matter injury development.Methods A total of 152 Sprague-Dawley newborn rats(3 days old) were randomly divided into white matter injury group(n=76) or control group(n=76).The white matter injury model was established by ligation of the right common carotid artery and hypoxic exposure(8% O2 and 92%N2),and samples were collected at 3d,7d,14d and 21d after operation.Morphological changes of the brain tissues were observed under a light microscope,while myelination was analyzed using a transmission electron microscope.The expression and location of BMP4 and myelin basic protein(MBP),a marker for myelination,was detected by immunohistochemistry staining,expression levels of BMP4 and MBP proteins were analyzed by Western blotting,and BMP4 mRNA expression was measured by real-time PCR.Results Observed under the light microscope,the cellular structure was clear,fibers arranged closely and orderly in the white matter of the control group.Whereas in the white matter injury group,sparse cells,loose mesh shaped white matter,and disorderly oriented fibers were observed.In the control group,myelin sheath had regular morphology,uniform density,and same thickness,observed using the transmission electron microscope.While in the white matter injury group,the myelin sheath was loosened,thinned,lamellar separated,and boundary obscured.Using immunohistochemistry staining,Western blot,and real-time PCR analyses,it was found that the protein and mRNA expression of BMP4 had no significant change with the increase of age in the control group,while it was rapidly increased with the extending of ischemic time in the white matter injury group.Comparing with the control group,the expression of BMP4 was significantly increased since 3d after operation in the white matter injury group(P<0.05),and the difference between two groups became more significant with the extending of ischemic time.The expression of MBP protein was analyzed by immunohistochemistry staining and Western blot,and a gradual increase was found in both groups with the increase of age.However,the expression of MBP protein was significantly decreased on 14d and 21d after operation in the white matter injury group compared with the control group(P<0.05).Conclusion Myelination disorders exists in white matter injury induced by ischemia-anoxemia.Meanwhile,the expression of BMP4 is significantly increased in the white matter injury group,indicating a possibility that BMP4 involves in the regulation of myelination disorders in white matter injury.

Effects of perinatal exposure to nonylphenol on delivery outcomes of pregnant rats and inflammatory hepatic injury in newborn rats

The current study aimed to investigate the effects of perinatal exposure to nonylphenol (NP) on delivery outcome of pregnant rats and subsequent inflammatory hepatic injury in newborn rats. The pregnant rats were divided into 2 groups: control group (corn oil) and NP exposure group. Thirty-four pregnant rats were administered NP or corn oil by gavage from the sixth day of pregnancy to 21 days postpartum, with blood samples collected at 12 and 21 days of pregnancy and 60 days after delivery. The NP concentration was measured by HPLC, with chemiluminescence used for detection of estrogen and progesterone levels. Maternal delivery parameters were also observed. Liver and blood of the newborn rats were collected and subjected to automatic biochemical detection of liver function and blood lipid analyzer (immunoturbidimetry), and ultrastructural observation of the hepatic microstructure, with the TNF-α and IL-1β hepatic tissue levels evaluated by immunohistochemistry. Compared with the control group, the pregnant and postpartum serum NP and estradiol levels of the mother rats in the NP group were significantly increased, together with lowered progesterone level, increased number of threatened abortion and dystocia, and fewer newborn rats and lower litter weight. Serum and hepatic NP levels of the newborn rats measured 60 days after birth were significantly higher than those of the control group, as well as lower testosterone levels and increased estradiol levels. When observed under electron microscope, the hepatocyte nuclei of the control group were large and round, with evenly distributed chromatin. The chromatin of hepatocytes in the NP group presented deep staining of the nuclei, significant lipid decrease in the cytoplasm, and the majority of cells bonded with lysate. The results of immunohistochemistry showed that there was almost no TNF-α or IL-1β expression in the hepatocytes of the control group, while the number of TNF-α-, PCNA-, and IL-1β-positive cells in the NP group was increased, with higher integral optical density than the control group. Compared to the control group, the serum levels of alanine aminotransferase, aspartate aminotransferase, triglyceride and low-density lipoprotein in the newborn rats of the NP group were significantly increased. There was no significant difference in the serum level of high-density lipoprotein or cholesterol between the groups. Perinatal exposure to NP can interfere with the in vivo estrogen and progesterone levels of pregnant rats, resulting in threatened abortion, dystocia and other adverse delivery outcomes. High liver and serum NP levels of the newborn rats led to alteration of liver tissue structure and function. The NP-induced hepatotoxicity is probably mediated by inflammatory cytokines TNF-α and IL-1α.

Nerve protective effect of Baicalin on newborn HIBD rats

OBJECTIVES@#To investigate the nerve protective effect and mechanism of baicalin on newborn rats with hypoxic ischemic brain damage (HIBD).@*METHODS@#A total of 64 SD newborn rats were randomly divided into control group, model group, nerve growth factor group and baicalin group, with 16 in each group. Left carotid artery ligation method was adopted to establish the HIBD model except for in control group, which was treated with intraperitoneal injection of salin e10 mL/kg for 3 d. After oxygen recovery on hypoxia ischemia rats, intraperitoneal injection of saline 10 mL/kg was adopted in model group for 3 d. Intraperitoneal injection of nerve growth factor injection 50 μg/kg per day was adopted in nerve growth factor group for 3 d; intraperitoneal injection of radix scutellariae 16 mg/kg per day was adopted in baicalin group for 3 d after modeling. Four rats of each group were sacrificed at Day 1, 2, 3, 7 for microscopic observation of pathological morphological changes in brain tissue after HE staining, S-P immunohistochemical method was used for observation of Fas and FasL expression in brain cells.@*RESULTS@#Neat structure of cells was observed in control group; edema cells in disordered arrangement was observed in model group, with some cells necrosis and cavity change; tissue injury in nerve growth factor group and baicalin group was significantly lighter than that in model group; Fas and FasL expression in model group, nerve growth factor group and baicalin group were significantly higher than that in control group at different time points (P0.05).@*CONCLUSIONS@#Baicalin can reduce expression of Fas and FasL in HIBD rats, inhibit apoptosis of nerve cells, thus achieve the protective effect on HIBD rat nerves.

The effects of neural stem cell transplantation on Foxg1 gene expression in the subgranular zone of neonatal brain tissue damaged by hypoxia-ischemia / 中华物理医学与康复杂志

Objective To observe the effect on Foxg1 gene expression in the subgranular zone (SGZ) of cerebral tissue from neonatal rats with hypoxic-ischemic brain damage (HIBD) after transplantation of neural stem cells (NSCs) derived from umbilical cord blood.Methods Mononuclear cells separated from umbilical cord blood by density gradient centrifugation were cultured with orientated induction to differentiate the NSCs.The neuronal phenotype was identified using immunocytochemical methods.A total of 150 Sprague-Dawley rats were randomly divided into a sham-operation group,an HIBD group and an HIBD-NSCs group.Rats in the HIBD group and the HIBD-NSCs group were subject to ligation of the left carotid artery and then kept in a box under 8％ oxygen and 92％ nitrogen for 2.5 hours to establish the HIBD animal model.The artery was separated but not ligated in the sham operation group,which was not subjected to hypoxia.Twenty-four hours after the operation,the cultivated NSCs were transplanted by caudal vein injection into the rats in the HIBD-NSCs group.Rats were then sacrificed on the 3rd,7th,14th,21st and 28th days after the operation.Foxg1 gene expression in the SGZ was examined using in-situ hybridization methods.Results The number of Nestin-positive cells peaked on the 6th day of cultivation and then decreased by the 9th day.The Foxg1 gene was expressed in the SGZs of each group.The expression increased by the 3rd day after surgery in the HIBD and HIBD-NSCs groups,and peaked on 7th day after the operation,then declined gradually.The average expression level of Foxg1 in the HIBD group was significantly lower than that in the HIBD-NSCs group on the 7th day and thereafter.Conclusions Human umbilical cord blood mesenchymal stem cells can be induced and differentiated into neural stem cells.Foxg1 genes can still be present in the SGZ after birth.HIBD can induce the expression of Foxg1 genes.Transplanting NSCs can promote the expression of Foxg1 genes and improve morphological and functional recovery after HIBD,at least in neonatal rats.

Expression of cytoglobin gene in hypoxic-ischemic brain damage of newborn rat / 国际儿科学杂志

Objective To investigate the expression and change of cytoglobin(Cygb)gene on hypoxicischemic brain damage(HIBD).Methods Fifty SD rats of 7days old were divided into four study groups and one control group.The brain tissues were taken at 4hours,12hours,24hours and 48hours after the onset of HIBD.Cygb mRNA was determined by the reverse transcription PCR.One-way method of GraphPad Prism was used for statistics.Results The fragment length of PCR products was identical with experimental design.The expression level of Cygb gene increased at 4h after ischemia,and peaked at 24h.48h after HIBD,the Cygb gene level began to decrease.Conclusion The expression of Cygb in brain tissue increased rapidly after HIBD of newborn rats,suggesting that Cygb may have important function in the protection process of HIBD.

Culture of Motor Neurons from Newborn Rat Spinal Cord / 华中科技大学学报（医学）（英德文版）

ificantly longer than that in control group (P<0.05). This protocol can be adapted for various postnatal motor neurons studies.

Expressions of Glucose Transporter 1,3 Gene in Hippocampus of Newborn Rats with Hypoxic-Ischemic Brain Damage by Progesterone Intervention / 实用儿科临床杂志

Objective To observe the changes of expressions of glucose transporter 1,3 gene(GLUT1 mRNA and GLUT3 mRNA) in hippocampus after hypoxic-ischemic brain damage in newborn rats and effects of progesterone on them.Methods Forty SD rats(7-day-old) were divided randomly into 4 groups:normal group,sham operation group,hypoxic-ischemic group and progesterone group.Rats were subjected to right common carotid artery ligation and exposed to 80 mL/L oxygen and 920 mL/L nitrogen gas for 2 hours to establish hypoxic-ischemic model in hypoxic-ischemic group and progesterone group.The rats in sham operation group only received right common carotid artery ligation.Progesterone(8 mg/kg) or sesame oil(of same volume) was given intraperitonealy in progesterone group or other groups 30 minutes before operation.All rats were killed 24 hours after operations.The expressions of GLUT1 mRNA and GLUT3 mRNA in hippocampus of rats in every group were assessed by adopting RT-PCR technique.Results The expressions of GLUT1 mRNA and GLUT3 mRNA in the hypoxic-ischemic group(0.674?0.083,0.785?0.093) increased markedly compared with those in sham operated group(0.374?0.061,0.519?0.060)(Pa0.05).Conclusions Progesterone maintain the energy supply of the brain by up-regulating the expression of GLUT1 mRNA and GLUT3 mRNA and accelerating the transportation of glucose into brain,which may be one of the protective mechanisms.

Effects of Folate Deficiency during Pregnancy of Rats on Expression of Foliate Binding Proteins Gene and WNT Signal Transduction Pathway in Heart of Offspring / 实用儿科临床杂志

Objective To study the expressions of foliate binding protein 1(Folbp1),Wnt and ?-catenin genes on the heart of offspring during the development of embryo,whose mother was deficient of folic acid.Methods Control group involving 18 rats and study group involving 18 rats were choosen from the total 36 adult female SD rats randomly copulate with the male normal rats after feeding different fodder for 2 weeks.The heart of the 13.5,17.5 days embryos and the newborns were obtained.The expressions of Folbp1,Wnt and ?-catenin genes mRNA at the 3 periods were evaluated by RT-PCR.Results The expressions of Folbp1,Wnt and ?-catenin genes mRNA of the study group were significantly weaker than those of the control group in heart of the 13.5,17.5 days embryos and the newborns(all P

Effects of Erythropoietin on Activation of Cas pase-3 in Newborn Rat Hippocampal CA1 Region Pest Hypoxic-ischemic Brain Damage / 中华围产医学杂志

ObjectiveTo investigate the effects of erythro po ietin on prevention hypoxic-ischemic brain damage (HIBD) and activation of Casp ase-3 in Hippocampal CA1 region in newborn rats. Methods7 d Sprague-Dawley rats were divided into hypoxic-ischemic (HI) group (n=11), recombinant human erythropoietin (rhEPO) treated group (n=11), and sham-op erated control group (n=9). HIBD was established in both HI group and rhEPO treated group. The number of rats animals with spontaneous left-turn in two gro ups was counted respectively at subsequent different time: 0, 6, 12 and 24 h. Th e expression and distribution of activated Caspase-3 was detected by immunohist ochemistry analysis. The positive cells were calculated in hippocampal CA1 regio n of every groups.ResultsTwo rats in HI group and one in rhE PO treated group died from continuous convulsion during hypoxia. all survival ra ts in up two groups had spontaneously left-turn Compared with HI group, the r ate of spontaneous left-turn was dramatically lower in rhEPO treated group (HI group vs rhEPO treated group, 1/10 vs 6/9, P=0.0198) at 24 h after hypox ia. The positive stained cells were distributed dispersively in the brain of con trol group, and more intensively in the hippocampus and cerebral cortex of the o ther two groups. In CA1 region, the number of positive cells in HI group, was si gnificant higher than that in control group ( 41.38 ?2.09 vs 10.52?2.70 , P

Expressions of glutamate and EAAC1 glutamate transporter in neonatal rat after hypoxia-ischemia and their changes after administrated ganglioside GM1 / 中国临床药理学与治疗学

AIM: To study the neuroprotective effect and possible mechanism of ganglioside GM1 on neonatal rats with hypoxic ischemic encephalopathy (HIE). METHODS: A rat model of neonatal HIE was established, then the pathological changes and expressions of glutamate and EAAC1 glutamate transporter in the brain tissues were investigated in different periods after hypoxia ischemia (HI) and the subsequent changes of the above results after GM1 administrated were studied too. RESULTS: The damages of the brain by exposed to HI were alleviated remarkably after GM1 administrated. The levels of glutamate neuron expressions in the brain tissue decreased after HI but EAAC1 increased. GM1 could partly prevent glutamate neuron reduction induced by HI and increase EAAC1 expression. CONCLUSION: GM1 may have some protective effects on glutamate neuron in neonatal HIE, and the possible mechanism is related to the partial increasing of EAAC1 expression.

Effects of Folate Deficiency during Pregnancy on Expression of NKx2.5 Gene in Heart of Offspring / 实用儿科临床杂志

Objective To study the expression of NKx2.5 on the heart of offspring during the development of embryo,whose mother is deficient of folic acid.Methods 1.Control group involving 18 rats and study group involving 18 rats were chosen from the total 36 adult female SD rats randomly copulate with the male normal rats after feeding different fodder for 2 weeks.The heart of the 13.5 days,17.5 days embryos and the newborns were obtained;2.the expression of NKx2.5mRNA by RT-PCR was observed;3.the expression of NKx2.5 protein by Western-blotting was investigated.Results 1.The expression of NKx2.5 mRNA of study group was weaker than control group in heart of the 13.5 days,17.5 days embryos and the newborns(P