血小板激因子及其受体拮抗剂对创伤性脊髓水肿的影响

采用猫鞘内注射血小板激活因子（ＰＡＦ）及静脉注射ＰＡＦ受体拮抗剂的ＢＮ５２０２１观察其对猫脊髓损伤后脊髓水肿的影响。结果显示，ＰＡＦ能够增加损伤后脊髓组织含水量及Ｎａ＾＋、Ｃａ＾２＋含量；降低Ｋ＾＋、Ｍｇ＾２＋含量；ＢＮ５２０２１能够降低伤后脊髓组织含水量，维持脊髓组织Ｎａ＾＋、Ｋ＾＋、Ｃａ＾２＋、Ｍｇ＾２＋含量的相对稳定，提示ＰＡＦ及其受体在创伤性脊髓水肿的病理生理过程中起重要作用，而ＰＡＦ受体     

钙离子在脑外伤后继发性肺损害中的作用

目的探讨钙离子（Ｃａ２＋）在脑外伤后继发性肺损害中的作用。方法用自由落体致大鼠脑外伤模型，伤后于５个不同的时相点，分别测定脑细胞、肺混合细胞内游离钙，并取标本作病理检查。结果伤后６小时，脑细胞及肺混合细胞内游离钙显著升高，伤后７２小时达到高峰，两者的变化呈直线正相关。病理检查结果显示伤后７２小时，脑、肺组织病理损害最为严重。结论脑外伤后，Ｃａ２＋在继发性肺损害中起着重要作用。     

钙离子通道拮抗剂对颅脑撞击伤后神经元钙通道电流的影响

目的：研究颅脑撞击伤后皮层神经元钙离子通道电流变化及钙离子通道拮抗剂对该电流的影响．方法：采用自由落体撞击装置对大鼠大脑左顶部进行硬膜外撞击致伤．伤后分离出损伤灶边缘区及健侧相应部位神经元，用膜片钳技术的全细胞方式记录神经元钙离子通道电流．结果：伤侧Ｃａ２＋电流（ＩＣａ）为－２４０±４６ｐＡ，较健侧－１４３±３８ｐＡ平均增加６８．５％．分别用１０μｍｏｌ／Ｌ尼莫地平灌流后，伤侧下降至－１０８±３７ｐＡ，下降率为５５．０％，健侧下降至－１１２±３５ｐＡ，下降率为２１．６％．结论：脑创伤后Ｃａ２＋内流增加，可能为细胞内Ｃａ２＋超载的主要原因之一．尼莫地平对创伤后神经元Ｃａ２＋电流的抑制作用较正常神经元更明显，对阻止Ｃａ２＋内流，减轻神经细胞损害有一定作用．     

高温,噪声及复合因素对大鼠脑线粒体Ca^2＋,Mg^2＋含量的影响

为研究高温、噪声对大鼠脑皮层线粒体Ｃａ＾２＋、Ｍｇ＾２＋含量的影响，用原子吸收分光光度计进行测定。结果发现：（１）高温组脑线粒体Ｃａ＾２＋明显升高，与对照组、噪声级及复合组比较均有显著性差异（Ｐ＜０．０１）；（２）复合组脑线粒体Ｃａ＾２＋明显减少，与对照组比较有显著性差异（Ｐ０．０５）；（３）高温组、噪声组及复合组与对照组比较，脑线粒体Ｍｇ＾２＋无显著变化，但复合组Ｍｇ＾２＋明显高于噪声线（Ｐ＜０     

尼莫地平对脑损伤后神经细胞钙通道和超微结构的改变

采用细胞内Ｃａ２＋荧光探针Ｆｕｒａ－２／ＡＭ检测大鼠脑损伤后神经元突触体胞浆中游离Ｃａ２＋浓度（［Ｃａ２＋］ｉ），以研究神经细胞钙通道变化，并选用Ｃａ２＋通道阻断剂尼莫地平治疗，观察神经细胞Ｃａ２＋通道变化和超微结构的改变。结果表明，脑损伤后神经细胞钙通道开放，突触体胞浆中游离［Ｃａ２＋］ｉ在伤后０．５小时即已升高为１．０９±０．０８×１０－６Ｍ／Ｌ水平（Ｐ＜０．００１），伤后６、２４、４８和７２小时，［Ｃａ２＋］ｉ持续处于１０－６Ｍ／Ｌ水平以上。同时发现神经细胞Ｃａ２＋超载时，其超微结构损害。应用尼莫地平治疗后，神经细胞胞浆游离［Ｃａ２＋］ｉ明显下降，超微结构损害显著减轻。     

脑缺血再灌流时〔^3H〕—三磷酸肌醇放射活性及突触体游离Ca^2…

研究大鼠脑血再灌流时〔＾３Ｈ〕－ＩＰ３放射活性及突触体（Ｃａ＾２＋）ｉ的变化，结果示缺血１ｍｉｎ就启动了肌醇脂质信使系统，引起〔＾３Ｈ〕－ＩＰ３显著增高，缺血２０ｍｉｎ突触体（Ｃａ＾２＋）ｉ增高，且持续至再灌流７ｄ，磷脂酶Ｃ抑制剂ＰＭＳＦ治疗能显著地抑制突触体（Ｃａ＾２＋）ｉ的升高，减轻海马ＣＡ１区缺血性神经元损伤。     

加压素在脑缺血再灌流海马迟发性神经元损伤中的作用

用沙土鼠制作迟发性神经元损伤动物模型。结果表明，脑缺血再灌流６－９６小时，海马ＣＡＩ区ＡＶＰ含量以及再灌流２６－９６小时的ＣＡＩ区含水量持续性增高。向侧脑室注射ＡＶＰ后，再灌流９６小时的海马ＣＡＩ区含水量和Ｃａ＾２＋浓度均较生理盐水对照组显著增加，而Ｎａ＾＋，Ｋ＾＋－ＡＴＰ酶活力却明显降低；侧脑室注射ＡＶＰ抗血清后，海马 ＣＡＩ区含水量和Ｃａ＾２＋浓度较对照组显著降低。     

回转器旋转对鸡胚脑细胞内游离Ca^2＋水平的影响及其恢复

探讨了不同胚龄鸡胚旋转不同时间后，脑细胞内游离Ｃａ＾２＋水平的变化，利用回转器模拟微重力生物效应及ｆｕｒａ－２比例荧光法，测定了孵化第６至１８ｄ鸡胚脑细胞游离Ｃａ＾２＋的浓度。结果显示：孵化第８至１７ｄ鸡胚经旋转不同时间后，其脑细胞内游离Ｃａ＾２＋浓度均有所下降其中１０ｄ鸡胚旋转４或７ｈ及第１３ｄ鸡胚旋转２４ｈ后，脑细胞内Ｃａ＾２＋浓度比对照组显著降低（Ｐ〈０．０１），第１０ｄ和１３ｄ鸡胚分别旋转     

中长期模拟失重大鼠骨骼肌线粒体钙含量和肌浆网Ca^2＋—A…

为探讨失重对肌肉功能的影响，观察了中长期模拟失重时大鼠骨骼肌细胞内Ｃａｗ＾２＋转运功能变化，测定了比目鱼肌，腓肠肌线粒体钙含量和肌浆网（ＳＲ）Ｃａ＾２＋－ＡＴＰａｓｅ活性。结果是悬吊１５、３０ｄ大鼠比目鱼肌和腓肠肌线粒体Ｃａ＾＋２含量增加，肌浆网Ｃａ＾２＋－ＡＴＰ酶活性降低，说明中长期模拟失重肌细胞内Ｃａ＾２＋转运功能发生了改变，这可能是影响肌肉收缩特性的因素之一。     

模拟失重大鼠心肌肌浆网Ca^2＋摄取功能的变化

为研究模拟失重对大鼠心肌肌浆网Ｃａ＾２＋摄取功能的影响，采用差速离心法制备心肌肌浆网（ＣＳＲ）精制膜，测定ＣＳＲ囊泡膜各种ＡＴＰ酶活性，并用Ｍｉｌｌｉｐｏｒｅ滤过技术测定ＣＳＲ囊泡的Ｃａ＾２＋摄取功能。结果表明，４周模拟失重大鼠心肌肌浆网膜Ｃａ＾２＋，Ｍｇ＾２＋－ＡＴＰ酶活性不变，Ｃａ＾２＋－激活ＡＴＰ酶活性却较对照组降低６．８％（Ｐ＜０．０５）。模拟失重组心肌肌浆网囊泡ＡＴＰ依赖性Ｃdisplay stat     

颅脑创伤早期兴奋性氨基酸的变化及其意义

目的探讨兴奋性氨基酸（ＥＡＡ）在颅脑创伤早期的变化及钙通道阻断剂治疗外伤性脑水肿的机制。方法通过大鼠脑外伤模型检测了外伤后脑组织含水率、脑脊液中ＥＡＡ含量以及钙通道阻断剂（尼莫地平）对两者变化的影响；通过神经细胞培养模型观察了钙通道激动剂（Ｂａｙ－Ｋ－８６４４）、谷氨酸（Ｇｌｕ）、天门冬氨酸（Ａｓｐ）及尼莫地平对神经元钙通道电流（ＩＣａ）的影响。结果外伤后脑组织含水率、脑脊液ＥＡＡ含量均较对照组明显增加（Ｐ＜０．０５），用尼莫地平治疗后两者均明显下降（Ｐ＜０．０５）。Ｂａｙ－Ｋ－８６４４、Ｇｌｕ、Ａｓｐ可使培养神经元ＩＣａ增加，并存在明显的剂量依赖关系。结论颅脑创伤早期ＥＡＡ释放增加，ＥＡＡ促进Ｃａ２＋内流。钙通道阻断剂治疗外伤性脑水肿的机制是抑制脑外伤后ＥＡＡ释放及直接减少Ｃａ２＋内流。     

冷冻伤性脑水肿的发生机制及尼莫地平治疗作用的实验研究

目的研究大鼠冷冻伤性脑水肿不同时间脑组织伊文思兰（ＥＢ）、突触体内［Ｃａ２＋］ｉ及Ｃａ２＋－ＡＴＰ酶活性变化与脑含水量变化之间的规律，以探讨冷冻伤性脑水肿的发生机制和类型。方法干湿法测定脑组织水分含量，甲酰胺法测定ＥＢ含量，Ｆｕｒａ－２／ＡＭ荧光标记法测定突触体内［Ｃａ２＋］ｉ，微量定磷法测定线粒体Ｃａ２＋－ＡＴＰ酶活性。采用尼莫地平进行治疗，研究其对ＥＢ含量、［Ｃａ２＋］ｉ、Ｃａ２＋－ＡＴＰ酶活性和脑水肿的影响。结果冷冻伤后３０分钟即已发生Ｃａ２＋超载，伴随Ｃａ２＋－ＡＴＰ酶活性下降及脑组织水分含量及ＥＢ含量增加。尼莫地平治疗后ＥＢ含量和［Ｃａ２＋］ｉ明显下降，而Ｃａ２＋－ＡＴＰ酶活性明显恢复，脑水肿明显减轻。结论ＢＢＢ的通透性增加和细胞内钙通道开放，钙离子浓度超载在脑水肿的发生与发展过程中起了重要作用。冷冻伤性脑水肿在早期既有细胞毒性脑水肿，又有血管源性脑水肿，即混合性脑水肿。     

冷冻伤性脑水肿钙—ATP酶活性变化与脑水肿的关系

目的：研究大鼠冷冻伤性脑水肿后线粒体钙－ＡＴＰ酶活性变化与脑水肿的关系，同时应用尼莫地平干预，观察其对神经细胞钙－ＡＴＰ酶活性和脑水肿的影响．方法：测定线粒体内钙－ＡＴＰ酶活性，干湿法测定脑组织含水量的改变．结果：冷冻伤后３０分钟，线粒体钙－ＡＴＰ酶活性显著下降，４小时和２４小时组钙－ＡＴＰ酶活性持续进行性下降；相反脑组织水份含量则明显增高．应用尼莫地平干预，钙－ＡＴＰ酶活性明显回升，脑水肿则显著减轻．结论：冷冻伤后线粒体内钙－ＡＴＰ酶活性下降与脑组织水份含量增高密切相关，尼莫地平通过升高线粒体内钙－ＡＴＰ酶活性等，可明显减轻脑水肿     

Oxidant production and immune response after stretch injury in skeletal muscle.

PURPOSE: This study investigated oxidant production and associated immune response after acute muscle stretch injury. METHODS: A standardized single stretch injury was performed on the tibialis anterior (TA) muscle of 36 male New Zealand white rabbits while contralateral control limbs underwent a sham surgery. Animals were sacrificed 0, 4, 12, 24, 48, and 72 h after injury. Potential sites of oxidant production, measured with a dichlorofluorescein (DCF) probe, were evaluated using two separate buffers. RESULTS: Nonmitochondrial oxidant production measured under basal buffer conditions (0.1 M potassium phosphate) was increased in both injured and control limbs at 24 h (P < 0.01) and was greater in the injured limb at 12 and 48 h (P < 0.01). There was also an interaction of time and injury (P < 0.05). Maximum oxidant production by neutrophils and macrophages, stimulated by the induced buffer (including 1.7 mM ADP, 0.1 mM NADPH, 0.1 mM FeCl3), was increased in both injured and control limbs at 4 h (P < 0.01) and was greater in the injured limb at 48 h (P < 0.01). Myeloperoxidase (MPO) activity, indicating the presence of activated neutrophils, was higher in the injured limb at 4 and 48 h (P < 0.01). The activities of superoxide radical producing and quenching enzymes, xanthine oxidase (XO) and superoxide dismutase (SOD), were elevated at 24 (P < 0.01) and 4 h (P < 0.05), respectively, but showed no difference between injured and control limbs. CONCLUSION: We conclude that acute muscle stretch injury and the required surgeries to generate the injury result in a biphasic increase in oxidant production in both injured and control limbs, suggesting a systemic immune response. The increase in oxidant production at 4 h may be caused by an increase in activated neutrophils, whereas XO activity may contribute to oxidant generation at 24 h.     

脑复合剂对脑损伤大鼠Na+-K+-ATP酶、Ca2+-ATP酶活性及Ca2+调控的影响

目的 观察刨伤性脑损伤(traumatic brain injury,TBI)大鼠脑组织匀浆及线粒体Na+-K+-ATP酶、Ca2+-ATP酶活性、神经细胞内游离Ca2+浓度及脑组织钙调蛋白(CaM)表达的动态变化,探讨脑复合剂脑保护作用的分子生物学机制.方法 建立大鼠TBI模型,分别设立假手术组、TBI组及中药治疗组.中药治疗组给予脑复合剂10 g·kg-1·d-1,假手术组及TBI组给予同等剂量等渗盐水,2次/d,连续7 d.分别于TBI后24 h、72 h、1周等3个时相点处死大鼠,动态定量分析各组大鼠脑组织匀浆及线粒体Na+-K+-ATP酶、Ca2+-ATP酶活性、神经细胞内游离Ca2+浓度及脑组织CaM表达的变化.结果 TBI组各时相点大鼠脑组织匀浆及线粒体Na+-K+-ATP酶、Ca2+-ATP酶活性均明显降低,于TBI后72 h后逐渐恢复,1周时仍明显低于假手术组.中药治疗组大鼠脑组织Na+-K+-ATP酶、Ca2+-ATP酶活性显著增加(P＜0.05);TBI组各时相大鼠脑组织神经细胞内游离Ca2+浓度及脑组织CaM表达均有不同程度增高,于伤后24 h达高峰,持续至72 h仍高于假手术组.而中药治疗组各时相大鼠神经细胞内游离Ca2+浓度及脑组织CaM表达明显低于TBI组(P＜0.05).结论 脑复合剂的脑保护作用可能与增加脑组织Na+-K+-ATP酶、Ca2+-ATP酶的活性,从而减轻TBI后脑细胞的能量代谢障碍,降低神经细胞内游离Ca2+浓度及脑组织CaM表达,减轻Ca2+超载所导致的继发性脑损伤有关.     

Validation of sodium/glucose cotransporter proteins in human brain as a potential marker for temporal narrowing of the trauma formation

In many forensic cases, the existence of a traumatic brain injury (TBI) is an essential factor, and the determination of the survival time is nearly as important as the determination of whether or not a trauma exists. Since it is known that glucose uptake increases in injured brain cells in order to perpetuate the neuronal integrity, this study focuses on the pathomechanism of brain glucose supply via sodium/glucose cotransporters 1 and 2 (SGLT1, SGLT2) following traumatization. Human cerebrum tissue of male and female individuals who died following TBI was taken from the contusional and contralateral regions, as well as from individuals deceased due to cardiac arrest (control group). Total SGLT1 and SGLT2 protein expression was analyzed by immunoblotting comparing injured and non-injured tissue. The immunoreactivity in contusional cerebral cortex region began to increase 3 to 7 h following traumatization. We found that both SGLT1 and SGLT2 protein expression increased significantly 37 h post-injury compared to the control group. SGLT1 rose significantly at 52 h post-injury and peaked significantly at 72 h, while SGLT2 rose significantly at 52 and 72 h after injury. By compiling these data, we predict a standard operator via SGLT expression as a comparative expression assertion to determine post-injury survival time for unknown cases. Our result suggests that SGLT1 and SGLT2 protein expression may be useful in forensic practice as an effective target to analyze the existence of a TBI and to determine the time of the traumatization.

     

大鼠颅脑液压损伤后脑组织内Na^＋—K^＋—ATP酶活性的变化

目的：测定大鼠颅脑液压损伤后脑组织内Ｎａ＋－Ｋ＋－ＡＴＰ酶活性，探讨其在继发性脑损伤中的作用。方法：利用大鼠颅脑液压损伤模型，在致伤后６小时测定脑组织含水量、脑组织内Ｎａ＋－Ｋ＋－ＡＴＰ酶活性。结果：脑损伤６小时后，损伤侧脑组织内含水量明显增加（Ｐ＜０．０５），Ｎａ＋－Ｋ＋－ＡＴＰ酶活性明显下降（Ｐ＜０．０５）；而未损伤侧脑组织含水量和脑内Ｎａ＋－Ｋ＋－ＡＴＰ酶活性均无显著改变（Ｐ＞０．０５）。结论：脑损伤后脑组织内Ｎａ＋－Ｋ＋－ＡＴＰ酶活性下降是继发性脑损害发生和发展的重要因素之一。     

Effect of muscle stretching on the activity of neuromuscular transmission.

The purpose of this study was to investigate how muscle stretching affects the activity of neuromuscular transmission. The magnitude of the post-tetanic potentiation (PTP) of miniature end-plate potential (m.e.p.p.) frequency was measured in the rat soleus muscle at resting length and stretched length. The parameters of the magnitude of PTP can be indicators of the kinetics of Ca2+ metabolism in the nerve terminal. One of the parameters, normalized initial post-tetanic frequency (f), was significantly increased by stretching muscle 10% (P less than 0.05) and 20% (P less than 0.01) of its resting length. Another parameter, the time constant of augmentation (tau a), was not significantly changed by muscle stretching. The time constant of potentiation (tau p) was significantly increased by 20% muscle stretching (P less than 0.05). These results indicate that the Ca2+ conductance, especially the voltage-dependent Ca2+ influx, of the nerve terminal could be increased by muscle stretching. Greater Ca2+ conductance of the nerve terminal would increase intracellular free Ca2+. Consequently, the probability of transmitter release would be increased, because Ca2+ is closely related to the activity of the transmitter release mechanism.     

Calcium antagonistic effects of Chinese crude drugs: preliminary investigation and evaluation by 45 Ca.

Coronary and other diseases in cardiac or brain blood vessels are considered to be due to the excessive influx of Ca(2+) into cytoplasm. If Ca(2+) channels in cell membrane are blocked by medicines or other substances with considerable calcium antagonistic effects, these diseases might be cured or controlled. The influence of some Chinese crude drugs, including Crocus sativus, Carthamus tinctorius, Ginkgo biloba and Bulbus allii macrostemi on Ca(2+) influx in isolated rat aortas was investigated by using (45)Ca as a radioactive tracer, and their calcium antagonistic effects were evaluated. It can be noted that Ca(2+) uptake in isolated rat aorta rings in normal physiological status was not markedly altered by these drugs, whereas the Ca(2+) influxes induced by norepinephrine of 1.2 micromol/L and KCl of 100 mmol/L were significantly inhibited by Crocus, Carthamus and Bulbus in a concentration-dependent manner, but not by Ginkgo. The results show that extracellular Ca(2+) influx through receptor-operated Ca(2+)channels and potential-dependent Ca(2+)channels can be blocked by Crocus, Carthamus and Bulbus. This implies that these Chinese crude drugs have obvious calcium antagonistic effects.