Research on the Mechanism of Neuroprotection of Mild Hypothermia (35℃)-Combination Therapy with Neuroprotective Agents-

亚低温（35℃）神经保护机制研究-神经保护剂联合治疗-

基本信息

批准号：
14570624
负责人：
KATAYAMA Yasuo
金额：
$ 2.24万
依托单位：
Nippon Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570624/
关键词：
Hypothermia Rat Focal Ischemia Apoptosis Neuroprotection Immunocytochemistry In situ hybridization

项目摘要

The aim of this study is to determine whether neuroprotective agents (a selective thrombin inhibitor, argatroban, or immunosuppressant, FK506), would prevent neuronal cell death and whether extra-mildhypothermia (35℃) would enhance the neuroprotecive effect of a selective thrombin inhibitor following transient focal ischemia in rats. Sprague-Dawley rats were subjected to MCAo using an intraluminal suture technique for 2hrs. The rats were repeifused for 24h and decapitated for infarct and edema analysis. Argatroban-treated animals received a continuous injection of argatroban (3.0mg/kg) for 24 hrs by after the onset of ischemia, while vehide-treated groups received same dose of vehide. FK506-treated animals received a single injection of FK506 (0.3mg/kg) venously at 2 hours after isehemic induction, while vehide-treated groups received same dose of vehide. During ischemia, temporal muscle and rectal temperatures were monitored and maintained at 37℃ in the normothermic animals and at 35℃ … More in the hypothermic animals. Animals were randomly divided into the following four groups (each, n=6): (I) vehicle-treated normothermic group (control) at 37℃ (rectal and temporalis muscle temperatures); (II) argatroban or FK506-treated normothermic group at 37℃; (III) vehicle-treated hypothermic group at 35℃; (IV) argatroban or FK506-treated hypothermic group at 35℃. Temporal muscle and rectal temperatures were maintained during ischemia at 37±0.2℃ (normothermic groups) or 35±0.2℃ (hypothermic groups). Argatroban (162±28mm^3) ameliorated the cortical ischemic damage compared with the control (205±55mm^3) significantly (p<0.05). Moreover, argatroban with mild hypothermia decreased the cortical infarct volume (114±28mm^3) significantly compared with those of groups I and III(170±27mm^3) (p<0.05). Furthermore, argatroban with mild hypothermia also decreased the cortical edema (29±11mm^3) volume significantly compared with those of groups I, II and III (68±23mm^3、52±13mm^3、61±16mm^3) (p<0.05). In the borderzone of cortex and striatum, argatroban reduced expression of the proapoptotic Bax protein, whereas increased upregulation of antiapoptotic protein Bcl. Moreover, TUNEL positive cells in the borderzone were decreased in the groups treated by argatroban Argatroban improved neurological symptoms significantly (p<0.05) and also improved survival rate. The combination of FK506 and mild hypothermia significantly reduced infarct volume (cortex,-61%;striatum,-31%) and edema volume (cortex,-57%;striatum,-41%), while mild hypothermia or FK506 alone failed to improve ischemic brain damage. These results demonstrate that extra-mild hypothermia (35℃) enhances neuroprotective effects of neuroprotective agents (a selective thrombin inhibitor, argatroban or immunosuppressant, FK506), suggesting that this combined therapy, extra-mild hypothermia (35℃) plus neuroprotective agents, may be a new therapeutic strategy for the treatment of acute stroke. Less

这项研究的目的是确定神经保护剂（选择性凝血酶抑制剂，Argatroban或免疫抑制剂FK506）是否会防止神经元细胞死亡以及外旋化性疗法（35℃）是否会增强选择性溶神经药物的选择性溶神经抑制剂抑制剂群体的神经药物效应。 Sprague-Dawley大鼠使用2小时的内部缝合技术对MCAO进行MCAO。将大鼠重复24小时，并斩首以进行感染和水肿分析。经过局部缺血发作后，经过阿atroban治疗的动物不断注射Argatroban（3.0mg/kg），持续24小时，而经汽车处理的组则接受了相同剂量的蔬菜。受FK506治疗的动物在等性诱导后2小时加热地注射了FK506（0.3mg/kg），而车辆治疗的组则接受了相同剂量的蔬菜。在缺血期间，临时肌肉和直肠温度被监测并保持在常规动物中的37℃，而在低温动物中则为35℃…更多。将动物随机分为以下四组（每组n = 6）：（i）在37℃（直肠和临时肌肉温度）处进行了媒介物处理的正常温度组（对照）；（ii）Argatroban或FK506处理的正常热群，位于37℃；（iii）在35℃的车辆处理的低温组；（iv）Argatroban或FK506处理的低温组为35℃。在缺血期间，颞肌和直肠温度保持在37±0.2℃（拟态度组）或35±0.2℃（低温组）。与对照（205±55mm^3）相比，Argatroban（162±28mm^3）改善了皮质缺血损伤（P <0.05）。此外，与I和III组（170±27mm^3）相比，具有轻度体温过低的Argatroban显着降低了皮质梗塞体积（114±28mm^3）（P <0.05）。此外，与I，II和III组相比，与温和体温过低的Argatroban还显着降低了皮质水肿（29±11mm^3）的体积（68±23mm^3，52±13mm^3，61±13mm^3，61±16mm^3）（p <0.05）。在皮质和纹状体的边界区域中，Argatroban降低了促凋亡Bax蛋白的表达，而抗凋亡蛋白BCL的上调增加。此外，在Argatroban Argatroban治疗的组中，边界区域中的TUNEL阳性细胞得到了改善，可显着改善神经系统症状（P <0.05），并提高了存活率。 FK506和温和体温过低的组合显着降低了梗塞量（皮质，-61％；纹状体，-31％）和水肿量（皮质，-57％；纹状体，-41％），而轻度低温或FK506仅无法改善缺血性大脑损伤。这些结果表明，超级体温过低（35℃）增强了神经保护剂的神经保护作用（一种选择性凝血酶抑制剂，Argatroban或免疫抑制剂，FK506），这表明这种合并治疗，额外的低温（35℃）以及神经保护性的策略是一种新的治疗疗法，可以是新的。较少的