缺氧缺血性脑病新生鼠脑细胞与红细胞内钙相关性研究

缺氧缺血性脑病新生鼠脑细胞与红细胞内钙相关性研究李着算农绍汉冯泽康１９９３年７月～１９９４年２月利用３０只缺氧缺血性脑病（ＨＩＥ）新生鼠的动物模型，研究了ＨＩＥ对红细胞内与脑细胞内的钙变化之间的相关关系，以探讨ＨＩＥ时红细胞内钙的变化是否可以反映脑细...     

新生儿缺氧缺血性脑病并低血钙时能否补钙问题的探讨

缺氧缺血性脑病新生儿（ＮＨＩＥ）并低钙血症量补钙后是否会加重细胞内钙超载是临床上急待解决的问题，本研究了ＮＨＩＥ的红细胞胞浆游离钙（ＲＢＣ（Ｃａ＾２＋）ｉ）红细胞总钙（ＲＢＣ（Ｃａ＾２＋）Ｔ）及血浆总钙（Ｐ（Ｃａ＾２＋）Ｔ）的动态变化，对ＮＨＩＥ伴低钙血症者测定补钙前后ＲＢＣ（Ｃａ＾２＋）ｉ，ＲＢＣ（Ｃａ＾２＋）Ｔ）及Ｐ（Ｃａ＾２＋）Ｔ）的动态变化，对ＮＨＩＥ伴低钙血症者测定补钙前后ＲＢＣ（Ｃａａ     

尼莫地平对新生猪缺氧缺血性脑损伤的防治作用

为探讨莫尼地平对新生儿缺氧缺血性脑病的防治作用，将８６只新生猪分为正常对照组，缺氧缺血性脑损伤（ＨＩＥ）１小时（１ｈｒ）、２４ｈｒ、７２ｈｒ组，尼莫地平０．５μｇｋｇ－１ｍｉｎ－１治疗后１ｈｒ、２４ｈｒ、７２ｈｒ组，尼莫地平１μｇｋｇ－１ｍｉｎ－１治疗后１ｈｒ、２４ｈｒ、７２ｈｒ组。检测各组红细胞胞浆总钙（ＲＢＣＴＣａ）及游离钙（ＲＢＣＣａ２＋ｉ）的浓度变化。结果：ＨＩＥ１ｈｒ组ＲＢＣＴＣａ和Ｃａ２＋ｉ均明显高于正常组对照（Ｐ＜０．０１），随着缺氧缺血状态的改善，ＲＢＣＴＣａ和Ｃａ２＋ｉ含量逐渐下降，ＨＩＥ７２ｈｒ组接近正常对照组（Ｐ＞０．０５）。两种剂量尼莫地平各治疗组ＲＢＣＴＣａ和Ｃａ２＋ｉ均明显低于ＨＩＥ１ｈｒ和２４ｈｒ组（Ｐ＜０．０１），与正常对照组比较，差异无显著意义（Ｐ＞０．０５）。提示：窒息后细胞内钙的沉积与新生猪ＨＩＥ的发生有关，尼莫地平能阻止Ｃａ２＋内流，有效地保护神经细胞。     

新生儿缺氧缺血性脑病血清神经元特异性烯醇酶与血浆内皮素—1…

为探讨血清神经元特异性烯醇酶（ＮＳＥ）、血浆内皮素－１（ＥＴ－１）与新生儿缺氧缺血性脑（ＨＩＥ）的分别用酶联免疫法和放射免疫法测定２５例正常足月新生儿和５９例足月ＨＩＥ新生儿血清中ＮＳＥ浓度和血浆ＥＴ－１浓度，并观察其动态变化。结果ＨＩＥ中，、重度组生后３天内ＮＳＥ显著高于对照组和轻度组；第７生逐渐下降，但重度组高于轻、中度组和对照组。ＥＴ－１浓度于生后３天内ＨＩＥ组显著高于对照组，且重度组高于中     

超氧化物歧化酶和维生素E对新生大鼠缺氧缺血脑组织脂质过氧化物的影响

本研究通过给新生缺氧缺血脑病（ＨＩＥ）大鼠腹腔注射不同剂量的超氧化物歧化酶（ＳＯＤ）、维生素Ｅ（ＶｉｔＥ）维生素Ｃ（ＶｉｔＣ），观察这些药物对ＨＩＥ大鼠脑组织脂质过氧化物（ＬＰＯ）的影响；分析用于大剂量的ＳＯＤ和ＶｉｔＥ＋ＶｉｔＣ时，ＨＩＥ大鼠脑内ＬＰＯ水平与仅注射生理盐水组相比地增加（ＳＯＤ大剂量组）。当将ＳＯＤ和ＶｉｔＥ＋ＶｉｔＣ剂量分别降低１０倍和２倍时，ＨＩＥ鼠脑ＬＰＯ水平明显降低并接近正     

新生儿缺氧缺血性脑病血浆血管活性肠肽的变化及纳络酮对其的影响

新生儿缺氧缺血性脑病 (ＨＩＥ)的基本病理生理过程是由于脑组织缺氧缺血和再灌注损伤造成的脑细胞损伤。动物实验证明急性脑损伤、脑缺氧时血浆及脑组织中血管活性肠肽 (ＶＩＰ)含量增高 ,对脑血管有调节作用[1,2 ] 。有关新生儿ＨＩＥ时ＶＩＰ的变化尚未见报道。本文通过观察ＨＩＥ新生儿血浆ＶＩＰ的变化及纳络酮对其的影响 ,探讨ＶＩＰ在ＨＩＥ发病中的病理生理意义 ,进一步了解纳络酮的作用机制。资料和方法一、一般资料　 1999年 7月～ 2 0 0 0年 12月北京医科大学第一医院儿科ＩＣＵ住院的ＨＩＥ新生儿 32例 ,其中 2 4例为ＨＩＥ治疗…     

心力衰竭患儿红细胞和心肌细胞内外钙的变化

为探讨心力衰竭（简称心衰）与心肌细胞钙代谢的关系，应用钙荧光指示剂Ｆｕｒａ－２和原子吸收分光光度法测定了１１例先天性心脏病合并心衰患儿心肌细胞和红细胞内外钙的浓度。结果发现，心衰时心肌细胞和红细胞内游离钙（２８１±４７ｎｍｏｌ／Ｌ，１．７６±０．０４Ｆ３３５／Ｆ３８５）较非心衰者（１２２±１３ｎｍｏｌ／Ｌ，１．４７±０．０８Ｆ３３５／Ｆ３８５）明显增多，红细胞总钙亦明显升高，但红细胞膜泵活性则较非心衰者下降；心衰纠正后，外周血红细胞内钙及膜泵活性恢复正常。提示，小儿心衰时心肌细胞内存在过多的钙积聚，且可能是心衰时心肌舒张功能障碍的原因之一。     

法乐四联症患儿心肌细胞和红细胞内外钙的变化

为了解法乐四联症（ＴＯＦ）患儿术前心肌舒张功能障碍的原因，采用钙荧光指示剂（Ｆｕｒａ－２）法和原子吸收分光光度（ＡＡＳ）法测定了１２例ＴＯＦ患儿心肌细胞、红细胞内外钙的变化。结果发现：ＴＯＦ术前心肌细胞游离钙（ＭｙｏＣａｉ）（３４７±８５ｎｍｏｌ／Ｌ）较先天性心脏病（简称先心病）组（１２２±１３ｎｍｏｌ／Ｌ）明显升高；红细胞游离钙（ＥｒｙＣａｉ）和红细胞总钙（ＥｒｙＣａｔ）（１．８９±０．５０Ｆ３３５／Ｆ３８５，２１．２±２．９μｍｏｌ／Ｌ红细胞）较先心病组（１．４７±０．０８Ｆ３３５／Ｆ３８５，１３．１±８．７μｍｏｌ／Ｌ红细胞）和健康组亦明显升高，而全血游离钙（ＢＣａｉ）和红细胞钠泵、钙泵活性明显下降。术后除钙泵活性外，ＢＣａｉ、ＥｒｙＣａｉ、ＥｒｙＣａｔ和红细胞钠泵活性均恢复正常。提示，钙内流增多和膜泵活性下降导致ＴＯＦ患儿心肌细胞和红细胞内存在过多钙积聚，可能是其术前舒张功能障碍的重要原因之一。     

新生儿窒息后血浆内皮素—1和前列环素水平变化的研究

应用放射免疫方法测定、研究了健康新生儿３３例和窒息新生儿５６例的血浆ＥＴ－１和６－Ｋｅｔｏ ＰＧＦ１α水平的动态变化。结果表明：（１）≤２天健康新生儿两种血浆浓度均显著高于生后８￣９组。（２）窒息新生儿两种血浆水平显著升高，与窒息的严重程度及窒息造成机体损害的严重度呈正相关；血浆ＥＴ－１与ＰＧＩ２处于动态平衡，表现了内皮细胞水平的自身调节。（３）重度ＨＩＥ血浆ＥＴ－１下降甚缓慢，与ＰＧＩ２处于失衡     

MRI在新生儿缺氧缺血性脑病中的应用

目的探讨新生儿窒息后所致缺氧缺血性脑病的脑损伤。方法对于４３例生后１０天内新生儿进行头颅ＭＲＩ检查，其中７例正常新生儿，３６例为缺氧缺血性脑病（ＨＩＥ）。结果根据ＭＲＩ影像特点将ＨＩＥ脑损伤分为：（１）脑实质水肿改变为主：①广泛脑水肿伴基底节区损伤：８／３６；②局灶性脑水肿伴基底节区损伤：１１／３６；③单纯性脑水肿：７／３６。（２）以脑白质改变为主：５／３６。（３）脑实质出血：４／３６，脑室旁梗塞继发出血１／３６，并且对１２例患儿进行随访，其中６例伴基底节区损伤患儿于６个月～１岁时ＭＲＩ仍可见异常Ｔ２低信号或Ｔ２高信号，１例脑室旁梗塞继发出血患儿于８个月时原发病灶呈Ｔ２高信号及Ｔ１低信号病灶信号。结论ＭＲＩ对基底节区微小病灶具有高度敏感性，有助于ＨＩＥ脑损伤的研究并指导治疗及评估预后     

三七皂甙治疗新生儿缺氧缺血性脑病的临床研究

目的：探讨三七皂甙对脑细胞的保护作用。方法：用新型钙染色剂Indo-1/Am检测缺氧缺血性脑病(HIE)患儿三七皂甙治疗前后细胞内钙变化,观察三七皂甙对细胞内钙超载的拮抗作用。将HIE患儿分为治疗组24例,对照组18例,同时设正常对照10例。在传统治疗的基础上,治疗组加用三七皂甙5～8 d,分别于入院时、治疗后24 h、72 h检测各组红细胞内游离钙(RBC[Ca2+]i)的浓度变化。结果：HIE治疗组及对照组RBC[Ca2+]i较正常足月儿升高,差异有非常显著性意义(P<0.01),HIE治疗组与HIE对照组比较,RBC[Ca2+]i在治疗前无明显差异(P>0.05),治疗后随着缺氧缺血状态的改善,检测48 h、72 h RBC[Ca2+]i,治疗组RBC[Ca2+]i明显下降,差异有显著性意义(P<0.01),HIE治疗组自身比较,RBC[Ca2+]i在治疗72 h后明显下降,差异有显著性意义[(2.619±0.175)vs(2.358±0.280);P<0.01]。治疗组中枢性呼吸衰竭、循环不良、胃肠功能紊乱治疗的有效率分别为100.0％(5/5)、83.3％(20/24)、91.7％(22/24),对照组有效率分别为20.0％(1/5)、33.3％(6/18)、16.7％(3/18),(P<0.01)。结论：三七皂甙能缓解细胞内钙超载,保护脑组织,改善临床症状。     

Increased Plasma and Erythrocyte Selenium Concentrations but Decreased Erythrocyte Glutathione Peroxidase Activity after Selenium Supplementation in Children with Down Syndrome

ABSTRACT. An investigation was made of the activity of glutathione peroxidase (GSH-Px) in erythrocytes and the levels of selenium in plasma and erythrocytes before, during and after selenium supplementation in children with Down syndrome (DS). This subject is of interest since it has been suggested that selenium supplementation could enhance the GSH-Px activity in erythrocytes, probably leading to unproved protection against oxygen radicals, which might cause damage by lipid peroxidation, especially in the brain. Forty-eight children with DS were treated with selenium-rich yeast tablets (10 μg/kg body weight/day) for 6 months. The supplementation was well tolerated and no side effects were observed. Selenium supplementation resulted in increased concentrations of selenium both in plasma and erythrocytes, but decreased GSH-Px-activity in erythrocytes. Plasma and erythrocyte selenium levels had almost regained the initial values 12 months after termination of the supplementation. Erythrocyte GSH-Px activity, on the other hand, remained reduced and did not return to the presupplementation levels. Until we gain more knowledge about the biological functions of selenium in man and the role of oxygen metabolism in the development of presenile dementia in DS, universal selenium supplementation in DS patients cannot be recommended.     

FREE AND BOUND TRYPTOPHAN IN HUMAN PLASMA DURING THE PERINATAL PERIOD

Abstract. The concentration of tryptophan and the degree of binding of the amino acid to protein were examined in human plasma during the perinatal period. Both total and unbound (free) tryptophan were higher in cord vein plasma than in the maternal circulation, the concentration gradient being approximately 1: 2. The proportion of the total plasma tryptophan concentration that was not bound to protein was less in cord vein plasma than in the maternal circulation. After birth the proportion in infant plasma fell significantly. Both total and free tryptophan fell during the first 24 hours of postnatal life. Total tryptophan returned to the cord vein plasma level 6–8 days after birth whilst free tryptophan failed to increase during the period of the observations. In premature infants total and free tryptophan also declined in concentration 12–24 hours after birth, suggesting the phenomenon to be related to birth rather than to gestational age. Phenylalanine remained unchanged whilst tyrosine increased in concentration during the first 80 hours of postnatal life. Thus, the availability of tryptophan to the tissues appears to decline during the immediate postnatal period and the results suggest that the requirement for tryptophan during this time may exceed the supply from standard artifical milk preparations.     

新生大鼠缺氧缺血性脑病模型脑组织钙调蛋白测定及脑活素治疗作用

目的：研究脑组织中钙调蛋白(CaM)含量与新生大鼠缺氧缺血性脑病(HIE)发病机制的关系及脑活素的治疗作用。方法：在建立新生大鼠HIE模型的基础上,采用依赖环核苷酸的磷酸二酯酶(PDE)法检测103只大鼠(其中对照组15只,实验组88只分3组)脑组织CaM含量,结合病理切片和透射电镜观察脑组织病理改变,同时通过侧脑室及肌内两种途径给予脑活素治疗。结果：HIE组脑组织CaM含量均明显增高,24 h(521.27±46.04) μg/g脑组织;48 h(509.52±35.98) μg/g脑组织;72 h(421.05±31.81) μg/g脑组织,与正常组(187.63±54.22)比较差异有显著性(P<0.01);治疗组CaM含量均明显降低,侧脑室注射组：24 h(435.21±56.17) μg/g脑组织;48 h(260.38±32.43) μg/g脑组织;72 h(197.64±19.21) μg/g脑组织;肌内注射组：24 h(441.04±30.66) μg/g脑组织;48 h(305.39±32.99) μg/g脑组织;72 h(217.71±52.89) μg/g脑组织;与HIE组比较差异有显著性(P<0.01),接近正常组水平。在神经元变性、坏死和间质水肿等方面,治疗组明显轻于HIE组。结论：CaM含量异常升高在HIE的发病环节中起一定作用。脑活素治疗HIE的机制可能与其显著降低脑组织中CaM含量有关。     

PLASMA CALCIUM FRACTIONS IN NORMAL SUBJECTS FROM BIRTH TO ADULT AGES

A simple and rapid method to determine ultra-filtrable calcium in plasma is described in detail. Total calcium, ultrafiltrable calcium (Ca_uf), and protein-bound calcium in plasma are reported for 202 normal children (range 0–16 years of age) divided into 12 age groups, as well as for 93 healthy male blood donors. There was a decrease in plasma calcium and ultrafiltrable calcium after 1–21 hours of life. The minimum value was reached at 24–48 hours of life. An increase was then seen during the first week of life. Later in childhood total calcium and Ca_uf decreased continuously and significantly with age, and with the most marked rate during the first two-three years. The ultrafiltrable calcium fraction was found to be responsible for the changes in total plasma calcium. In 24–48 hours of life and 5–7 days of life protein-bound calcium was significantly lower as well. There were no significant differences between males and females in childhood.     

Increased plasma and erythrocyte selenium concentrations but decreased erythrocyte glutathione peroxidase activity after selenium supplementation in children with Down syndrome

An investigation was made of the activity of glutathione peroxidase (GSH-Px) in erythrocytes and the levels of selenium in plasma and erythrocytes before, during and after selenium supplementation in children with Down syndrome (DS). This subject is of interest since it has been suggested that selenium supplementation could enhance the GSH-Px activity in erythrocytes, probably leading to improved protection against oxygen radicals, which might cause damage by lipid peroxidation, especially in the brain. Forty-eight children with DS were treated with selenium-rich yeast tablets (10 micrograms/kg body weight/day) for 6 months. The supplementation was well tolerated and no side effects were observed. Selenium supplementation resulted in increased concentrations of selenium both in plasma and erythrocytes, but decreased GSH-Px-activity in erythrocytes. Plasma and erythrocyte selenium levels but almost regained the initial values 12 months after termination of the supplementation. Erythrocyte GSH-Px activity, on the other hand, remained reduced and did not return to the presupplementation levels. Until we gain more knowledge about the biological functions of selenium in man and the role of oxygen metabolism in the development of presenile dementia in DS, universal selenium supplementation in DS patients cannot be recommended.     

缺氧缺血性脑病新生儿血浆肾上腺髓质素/内皮素-1值与血清神经元特异性烯醇化酶的相关性

目的 探讨缺氧缺血性脑病(HIE)足月新生儿血浆肾上腺髓质素(AM)/内皮素-1(ET-1)值的变化对血清神经元特异性烯醇化酶(NSE)的影响.方法 2005年1-12月收治HIE新生儿32例.按照HIE的诊断标准分为轻度组18例,中重度组14例.本院同期出生的30例健康新生儿(生后24 h)为健康对照组.采用放射免疫法动态测定HIE新生儿生后1、3、7 d及健康新生儿生后1 d血浆AM、ET-1和血清NSE水平.均数比较采用方差分析及S-N-K检验,双变量资料采用直线相关分析.结果 1.轻、中重度HIE组血浆AM、ET-1水平生后1、3、7 d均增高,与健康对照组比较,轻度组至3、7 d已无显著性差异(Pa＞0.05);中重度组至7 d仍有显著性差异(Pa<0.05).AM/ET-1值在3个时间点均较健康对照组减低,但仅中重度组生后1、3 d与健康对照组比较有统计学意义(Pa<0.05).2.HIE患儿血清NSE水平生后1、3 d增高,与健康对照组比较,仅中重度组有显著性差异(Pa<0.05);7 d,轻度组下降至健康对照组水平(P＞0.05),中重度组与健康对照组比较有显著性差异(P<0.05).且与AM/ET-1值呈显著负相关(r=-0.53 P<0.05).结论 AM和 ET-1共同参与HIE调节局部脑血流量及血脑脊液屏障功能的维护,AM/ET-1值测定对了解不同时期HIE患儿脑血流变化和脑损伤程度有重要意义.     

新生大鼠缺氧缺血性脑损伤NF-κBp65和TLR4表达变化探讨

目的 探讨核转录因子κBp65(nuclear factor-kappaBp65,NF-κBp65)和Toll样受体4(Toll-like receptor4,TLR4)在新生大鼠脑组织中的表达及其变化规律及相关性,研究两者在新生儿缺氧缺血性脑损伤发病机制中的作用.方法 出生7d新生大鼠随机数字表法分对照组和缺氧缺血组,制备新生儿缺氧缺血性脑损伤模型,并于缺氧缺血术后6h、12h、24h、72 h及7d取其脑组织制备光镜石蜡标本,观察脑组织的病理变化.采用免疫组织化学方法分析NF-κBp65和TLR4的表达.结果 新生大鼠缺氧缺血性脑损伤过程中,NF-κBp65和TLR4在神经元、小胶质细胞均有表达,以大脑皮质和海马部位表达变化明显;于缺氧缺血后6h NF-κBp65(0.2193 ±0.0247,0.2157 ±0.0304)和TLR4(0.3271 ±0.033 3,0.3039 ±0.0379)表达增加,24h NF-κBp65 (0.3564 ±0.023 5,0.3365 ±0.023 2)和TLR4(0.434 2 ±0.042 8,0.4193 ±0.0413)表达达到峰值,72 h NF-κBp65(0.289 2±0.032 0,0.2609±0.021 2)和TLR4(0.300 5±0.020 9,0.282 0±0.022 6)和7 d NF-κBp65 (0.2479±0.034 0,0.242 1 ±0.0254)和TLR4(0.2744 ±0.028 8,0.257 1 ±0.0275)表达下降.结论 NF-κBp65与TLR4表达呈正相关,提示可能在新生大鼠缺氧缺血性脑损伤中有相同机制.     

Novel treatments after experimental brain injury

Perinatal hypoxic-ischaemic encephalopathy(HIE) is being studied in laboratory models that allow the delayed cascade of events triggered by the energetic insult to be examined in detail. The concept of the ‘excitotoxic cascade’ provides a conceptual framework for thinking about the pathogenesis of HIE. Major events in the cascade triggered by hypoxia-ischaemia include overstimulation of N-methyl-D-aspartate type glutamate receptors, calcium entry into cells, activation of calcium-sensitive enzymes such as nitric oxide synthase, production of oxygen free radicals, injury to mitochondria, leading in turn to necrosis or apoptosis. New experimental approaches to salvaging brain tissue from the effects of HIE include inhibition of neuronal nitric oxide synthase, administration of neuronal growth factors, and inhibition of the caspase enzymes that execute apoptosis. Recent experimental work suggests that these approaches may be effective during a longer ‘therapeutic window’ after the insult, because they are acting on events that are relatively delayed. Application of modest hypothermia may allow these agents to be neuroprotective at even longer intervals after hypoxia-ischaemia.     

新生儿缺氧缺血性脑病血浆及脑脊液血小板活化因子的变化

目的　观察新生儿缺氧缺血性脑病 (HIE)血浆及脑脊液 (CSF)血小板活化因子 (PAF)的变化 ,探讨PAF在HIE发病中的作用。方法　于急性期和恢复期分别采集患儿血浆和CSF ,采用生物活性法检测血浆及CSF中PAF水平 ,并分析其与脑损伤和 1minApgar评分的关系。 结果　 1.HIE患儿急性期CSF中PAF水平明显高于对照组患儿 (P <0 .0 1) ,且与病情轻重呈明显正相关 (r =0 .6 5　P <0 .0 1) ;急性期中度与重度HIE患儿血浆PAF水平显著高于对照组 (P <0 .0 1) ,且与病情轻重呈明显正相关 (r =0 .5 9　P <0 .0 1) ,而轻度患儿与对照组无明显差异 (P >0 .0 5 ) ;恢复期与对照组比较无显著差异。 2 .血浆PAF水平与CSF中PAF水平呈正相关 ,血浆及CSF中PAF水平与脑损伤程度及 1minApgar评分呈明显正相关。 结论　PAF可能参与新生儿HIE的发病机制 ,CSF中PAF水平可作为判断HIE病情轻重和预后的重要指标