促血小板生成素对预测脑瘫高危儿的意义

目的探讨缺氧缺血性脑病(hypoxia-ischem ia encephalopathy,H IE)胎儿和H IE新生儿血清中促血小板生成素(Thrombopoietin,TPO)水平与脑损伤的关系,为脑瘫(cerebral palsy,CP)高危儿人群进行早期干预提供监测手段。方法收集23例H IE胎儿和34例H IE新生儿血清以及25例正常胎儿和30例正常新生儿血清,34例H IE新生儿包括11例轻度H IE,8例中度H IE和15例重度H IE。采用双抗体夹心ABC-ELISA法检测H IE胎儿组和轻、中、重H IE新生儿组血清中TPO的水平,并与正常胎儿组和正常新生儿组比较。结果H IE胎儿组和H IE新生儿组TPO分别高于正常胎儿组和正常新生儿组(分别P<0.01,P<0.01),重度H IE组TPO低于轻度H IE组(P<0.05)。结论血清TPO水平与H IE所致脑损伤严重程度有关。脐血TPO检测可为脑瘫高危儿人群进行早期干预提供监测手段。     

胰岛素样生长因子1对新生大鼠缺氧缺血性脑损伤保护机制的研究

目的　建立单侧缺氧缺血性脑损伤 (HIBD)动物模型 ,研究胰岛素样生长因子 1(IGF 1)对HIBD的影响和可能机制。　方法　选择健康 7日龄Wistar大鼠 12 0只 ,建立HIBD模型 ,随机分成假手术组、HIBD组、HIBD后 0 .2mg/kg人基因重组IGF 1干预组 (RH IGF 1组 )、0 .0 6 6mg/kg人基因重组IGF 1干预组 (SRH IGF 1组 )及盐水对照组 (对照组 )。各组按观察时段进一步分为 2 4、4 8、72h组 ,每组 8只。各组于规定时刻观测脑形态学改变、谷氨酸 (Glu)含量、凋亡细胞计数、Bcl 2蛋白表达。　结果　 (1)HIBD 4 8h组Glu(116 2 .2± 10 8.1)mg/kg ,较假手术组(75 0 .9± 5 3.4 )mg/kg明显升高 (P <0 .0 5 ) ;HIBD组凋亡细胞计数 [2 4h :(7.6± 1.9) % ,4 8h(12 .6±1.2 ) % ,72h :(13.8± 0 .9) % ],较假手术组 [2 4h(2 .0± 0 .2 ) % ,4 8h(2 .0± 0 .3) % ,72h(2 .0±0 .2 ) % ]明显增加 (P均 <0 .0 5 )。 (2 )与对照组相比 ,RH IGF 1组脑组织病变减轻 ;干预 4 8h组Glu[SRH IGF 1组 (781.4± 5 4 .2 )mg/kg ,RH IGF 1组 (74 0 .5± 4 6 .6 )mg/kg],较对照组 (112 6 .6± 4 8.0 )mg/kg明显降低 (P均 <0 .0 5 ) ;RH IGF 1组凋亡细胞计数 [2 4h :(3.6± 0 .9) % ,4 8h(8.2± 2 .2 ) % ,72h(9.4± 1.4 ) % ],较对     

羟丁酸钠对缺血缺氧性脑损伤新生大鼠大脑皮层神经细胞凋亡的影响

目的 观察羟丁酸钠对缺血缺氧性脑损伤新生大鼠大脑皮层神经细胞凋亡的影响。方法 新生7dSD大鼠随机分为假手术组（S组）、生理盐水对照组（C组）和羟丁酸钠组（71、坦和档组）。每组按Ⅲ（缺血缺氧）后不同时间点进一步分为1h、3h、1d、3d、7d5个亚组（n=6）。按Rice法制作缺血缺氧性脑损伤模型。C、γ1、γ2和γ3组HI后即腹腔分别注射生理盐水0．2ml／10g、羟丁酸钠50、100、200mg／kg，3次／日；S组术后吸空气2h，腹腔注射生理盐水0．2ml／10g，3次／日。TUNEL染色法检测HI后各时间点大脑皮层凋亡神经细胞。结果 HI后1h～7d，C、γ1、γ2、γ3组凋亡神经细胞数高于S组（P〈0．05），且于HI后1d达高峰；HI后3h-3d，C、γ3组凋亡神经细胞数多于γ1、γ2组（P〈0．05），而C组与γ3组之间凋亡神经细胞数差异无统计学意义（P〉0．05）；HI后7d，γ1组凋亡神经细胞数多于让组（P〈0．05）。结论 50、100mg／kg羟丁酸钠能抑制缺血缺氧性脑损伤新生大鼠大脑皮层神经细胞凋亡，且100mg／kg效果较好。     

Radioiodinated tracers for the evaluation of dopamine receptors in the neonatal rat brain after hypoxic-ischemic injury

In order to evaluate in vivo single-photon emission tomography (SPET) method of assessing cerebral function after hypoxic-ischemic injury in human neonates, we studied D₁ and D₂ dopamine receptors in a rat model. Seven-day-old rats underwent permanent unilateral common carotid ligation followed by exposure to 8% O₂. Two weeks later, in brains with no visible loss of hemispheric volume, striatal dopaminergic receptors were studied, with [¹²⁵I]TISCH and [1251]IBZM for the D₁ and D₂ dopamine receptors, respectively. Using [¹²⁵I]TISCH, we observed no modifications of D₁ receptors, but in contrast, ex vivo and in vitro autoradiographic experiments showed a 40% decrease in the striatal binding of [¹²⁵I]IBZM on both the ipsilateral and the contralateral side to the carotid ligation. These alterations were detected with IBZM, a D₂ dopamine receptor ligand usable for SPET imaging. Therefore, exploration of D₂ receptors by SPET in human neonates suffering from perinatal hypoxia-ischemia may be valuable for the diagnosis and follow-up of cerebral function damages. Correspondence to: D. Guilloteau     

新生鼠脑缺氧缺血时天冬氨酸半胱氨酸蛋白酶-3表达的研究

目的：探讨新生大鼠缺氧缺血脑损伤（HIBD）时皮质及海马中天冬氨酸半胱氨酸蛋白酶-3（caspase-3）表达的变化。方法： 分假手术组和模型（HI）组。用新生7 d的大鼠建立新生儿HIBD的模型，于缺氧后不同时点取脑，分别行病理学检查、免疫组化和半定量逆转录-聚合酶反应（RT-PCR），观察双侧皮质及海马caspase-3 蛋白及mRNA表达的程度。结果： HI组皮质及海马caspase-3 mRNA 于HI后0 h出现瞬时高表达，然后下降，6 h可检测到mRNA和蛋白同时升高，表达峰值于HI后24-48 h出现，然后下降。结论： Caspase-3参与未成熟脑HIBD后皮质及海马区神经细胞凋亡的进程。在其表达峰值出现以前，即HI后24-48 h之前，抑制caspase-3的表达，应是治疗脑损伤的适宜时机。     

White matter damage precedes that in gray matter despite similar magnetic resonance imaging changes following cerebral hypoxia-ischemia in neonatal rats

We hypothesized that the cerebral injury produced by hypoxia-ischemia (HI) in neonatal rats would differ in white compared with gray matter as detected histologically or with magnetic resonance (MR) imaging methods. Maps of T₂ and the apparent diffusion coefficient (ADC) of water were acquired in 1-week-old rats at times prior to cerebral HI (right carotid artery occlusion plus 1.5 h of hypoxia), within the last 5–10 min of HI, and 1 h or 24 h after HI. Near the end of HI, ADC decreased and T₂ increased in both cortical gray and subcortical white matter within the cingulum of the HI hemisphere. One hour after HI, ADC partially recovered, but T₂ remained increased and then increased further by 24 h post-HI. In contrast to the similar MR responses in white and gray matter, histological evidence for irreversible cell damage occurred in white matter earlier than in gray matter within the HI hemisphere. At 1 h post-HI, rarefied or disrupted nerve fibers and an increase in TUNEL-positive cells were observed within white matter in the cingulum, whereas neurons within the cortical gray matter appeared normal. By 24 h post-HI, damage was apparent in both white and gray matter. Thus, MR imaging detected acute tissue edema following cerebral HI in both gray and white matter but did not distinguish between the early irreversible tissue injury detected histologically in white but not gray matter in this rather severe model of neonatal encephalopathy.     

高压氧对新生大鼠缺氧缺血性脑损伤后在体神经干细胞的影响

目的： 新生鼠的神经生发区-室管膜下区（SVZ） 和海马齿状回（DG）聚集了多种不同发育阶段神经干细胞（NSCs），它们可分化产生新的神经元和胶质细胞。本研究探讨缺氧缺血性脑损伤（HIBD）对脑生发区NSCs的损伤及高压氧（HBO）对此损伤的影响。从在体NSCs 探讨HBO对新生大鼠HIBD的保护作用机制。 方法： 新生7 d龄SD大鼠随机分为4组：① 正常对照组（CON，n=16）；② HIBD 模型组（HIBD，n=16）；③ 高压空气组（HBA，n=16）；④HBO治疗组（n=16）。Rice法复制HIBD模型，并予HBA或HBO治疗，每天1次共7 d。BrdU免疫组化显示在体NSCs。并取损伤侧脑SVZ区组织，体外NSCs培养并进行神经干细胞球计数。 结果： HIBD组生发区在体BrdU 阳性细胞和体外培养的神经干细胞球数目明显少于对照组。HBO组SVZ区的BrdU阳性细胞增多；体外培养的神经干细胞球增多。HBA组增加不明显。 结论： 新生大鼠HIBD后生发区NSCs减少，HBO治疗可以减轻HIBD后NSCs的死亡。HBA治疗无明显作用。     

新生儿缺氧缺血性脑病血浆Th2 源细胞因子水平的研究

目的　研究新生儿缺氧缺血性脑病 (HIE)患儿辅助性T淋巴细胞亚群 2 (Th2 )细胞功能变化及其与HIE间的关系。　方法　采用ELISA法和单克隆抗体间接免疫荧光法 ,检测 4 6例HIE患儿不同病期血浆和外周血单个核细胞 (PBMC)体外分别产生白细胞介素 (IL) 6、IL 8、IL 10水平和T细胞IL 2受体 (IL 2R)表达率 ,并与正常足月新生儿脐血 (2 0例 )进行对照。　结果　HIE病程第 1天血浆及PBMC体外产生IL 6 [分别为 6 6 .3、80 4 .5ng L(中位数 ,以下同 ) ]、IL 8(4 10 .5、1983.6ng L)和IL 10 (2 7.9、14 0 .1ng L) ,明显高于正常脐血对照组 (分别为 1.9、187.7;91.0、76 6 .8;2 .6、16 .4ng L) (Z =4 .6 5 3～ 5 .85 6 ,P <0 .0 1) ;中重度组及并发多器官功能损害 (MOD)的HIE患儿血浆及PBMC体外产生IL 6 (分别为 86 .0、12 36 .4 ;111.3、174 6 .9ng L)、IL 8(分别为5 79.5、2 812 .2 ;5 32 .8、2 94 3.3ng L) ,分别高于轻度组 (分别为 5 2 .6、5 83.4 ;335 .3、15 2 8.9ng L)及无MOD组 (分别为 5 6 .6、713.8;35 2 .4、15 72 .4ng L)患儿 (Z =2 .2 2 8～ 2 .70 1,P <0 .0 5 ;Z =2 .877～4 .187,P <0 .0 1) ,而IL 10差异无显著性 (Z =0 .4 5 0～ 1.85 0 ,P >0 .0 5 ) ;T细胞IL 2R表达率 [(5 2 .8± 13.4 ) % ]明显     

脑红蛋白基因在新生大鼠缺氧缺血性脑损伤脑中的表达

目的　探讨缺氧缺血性脑损伤脑中脑红蛋白基因的表达及变化。　方法　随机将出生7d的Wistar大鼠 4 8只分成 6组 ,其中实验组在手术后制成脑损伤模型 ,并且在血流阻断 1、5、15、30、6 0min取脑 ;对照组为假手术组 ,术后即取脑。设计脑红蛋白基因引物 ,提取脑组织总RNA ,合成RT 单链cDNA做PCR反应。以Stata统计软件包的One way单因素方差分析进行数据统计处理。　结果　设计的新生大鼠脑红蛋白cDNA序列引物及扩增脑红蛋白编码区部分序列 ,其所得引物可靠、扩增产物与设计相符。检测中可见 :实验组脑红蛋白mRNA的表达在缺血 1min时 (A值 :1.2 36 )即快速增加达高峰 ,与对照组 (A值 :0 .6 4 1)相比 ,差异有非常显著性 (P <0 .0 1) ;5min时 (A值 :1.170 )仍然保持在高水平 (P <0 .0 1) ;15min时 (A值 :0 .4 98)迅速降低 ,与对照组相比 ,差异无显著性 (P >0 .0 5 ) ;此后脑红蛋白mRNA又见缓慢增高 (30min时脑红蛋白mRNAA值 :0 .6 4 2 ;6 0min时A值 :0 .6 17) ,与对照组相比 ,差异均无显著性 (P >0 .0 5 )。　结论　新生大鼠缺氧缺血性脑损伤后脑红蛋白mRNA表达增强 ,提示脑红蛋白可能在脑缺氧的适应性调节过程中起了重要作用。     

三日龄大鼠缺氧缺血性脑损伤对大脑MRI影像和学习记忆能力的远期影响

Objective To investigate the activation of apoptotic genes of the brain with hypoxia- ischemia (HI) in newborn SD rats, and MRI changes and memory and learning ability in adulthood. Methods HI was induced by right carotid artery ligation followed by 2.5 h of hypoxia (6% O2) on 3-day-old SD rats (n=36). Control pups were sham-operated (n = 27). Right brain hemisphere was collected at 12 h and 7 d after HI and subjected to an apoptosis Oligo GEArrayR. MRI and Morris water maze test were performed on both groups at 42 and 44 days old, respectively. Results Comparing to 12 h after HI, up-regulated apoptotic genes included TNF, Caspase and pro-apoptotit genes of Bcl2 families, whereas the anti-apoptotic genes of Bcl2 family were down-regulated at 7 d after HI. The MRI assessment of the rats in HI group demonstrated that the area of the right cerebra l cortex was significantly smaller than the left side and control [periventricular layer: (23.5±3.6) mm2 vs (33.0±4.3) mm2, (34.5±3.9) mm2; hippocampus layer: (18.9±4.4) mm2 vs (29.1±5.0) mm2,(30.8±4.5) mm2, both P<0.01]. During the navigation trial, the HI rats demonstrated longer escape latency (4th day: (52.7±35.9) vs (17.8±8. 9) s, P<0.01). HI rats passed the platform less times than the control ones (T= 292.5, P<0.05) in space probe trial. Conclusions The activation of apoptotic genes induced by HI brain injury remains until 7 days later, involving intrinsic and extrinsic apoptotic pathway. The apoptosis of neural cells may lead to poor development of the cortex and impair the memory and learning ability in the adult rats after neonatal hypoxia- ischemia injury.