经腹腔应用阿加曲班对脑出血后凝血酶神经毒性的抑制作用

Intraperitoneal Administration of Argatroban Inhibited Neurotoxicity of Thrombin in Perihemotoma in a Rat Model of Intracerebral Hemorrhage

目的:探讨经腹腔应用阿加曲班治疗脑出血(ICH)后凝血酶神经毒性损伤的可能性。方法:①研究阿加曲班对ICH及凝血酶注入后脑水肿、细胞损伤的影响。Wistar大鼠60只,随机分为假手术对照组(只进针不注血);ICH组(50μL自体尾血注入右尾状核);ICH+阿加曲班干预组(50μL自体尾血注入大鼠右侧尾状核,分别于术后3和12h经腹腔给予阿加曲班0.9mg·kg^(-1),总量0.6mL);凝血酶组(10U·2μL^(-1)凝血酶注入大鼠右侧尾状核);凝血酶+阿加曲班干预组(10U·2μL^(-1)凝血酶注入大鼠右侧尾状核,分别于术后3和12h经腹腔给予阿加曲班0.9mg·kg^(-1),总量为0.6mL)。各组均n=12,术后24h处死各组大鼠,每组中6只用于检测脑组织水含量,6只检测caspase-3免疫反应细胞及TUNEL阳性细胞数。②经腹腔注射阿加曲班对血肿容积的影响:建立胶原酶ICH大鼠模型组:注入Ⅰ型胶原酶+肝素;阿加曲班组:胶原酶ICH模型成功后3及12h分别经腹腔注入阿加曲班溶液0.9mg·kg^(-1),每次注入液体量为0.6mL,测定两组大鼠脑组织血肿血红蛋白的含量(测定血红蛋白A_(550)值)以评价阿加曲班对血肿容量的影响。结果:自体血ICH及凝血酶模型大鼠在阿加曲班干预后,ICH组及凝血酶组的TUNEL阳性细胞数、caspase-3阳性细胞数明显下降(P<0.01或P<0.05),脑组织水肿含量百分比明显降低(P<0.05)。胶原酶ICH血红蛋白A值为(0.45±0.12),阿加曲班干预组为(0.46±0.09),差异无统计学意义(P>0.05)。结论:ICH后3～12h经腹腔注入阿加曲班可减轻ICH后的脑水肿及细胞凋亡性损伤,且没有血肿增大的不良反应。

Aim：To investigate the possiblity of anti-neurotoxicity of thrombin therapy to perihematomal tissue injuries by intraperitoneal administration of argatroban in a rat intracerebral hemorrhage（ICH） model.Methods： Rats models were established by 50（XL autologous blood or 10 U thrombin injection into the right caudate nucleus. 0.9 mg·kg^（-1） argatroban was administrated intraperitoneally 3 h and 12 h after blood or thrombin injection.24 h later all rats were decapitated,then brain water content was measured and the expression of TUNEL positive cells or caspase 3 immunoreactivity（caspase-3IR） cells in perihematoma were detected.To verify whether hemorrhage volume could be increased by argatroban,hemoglobin content was measured with the model of cerebral hemorrhage induced by type I collagenase and heparin in the rats.Results：After argatroban administration, TUNEL positive cells,caspase-3IR cells,and brain water content were all decreased significantly in ICH groups and thrombin injection groups.The using of argatroban intraperitoneally did not change hematoma volume in ICH models.Conclusion：Thrombin takes a role in perihematomal tissue injuries in intracerebral hemorrhage.Cell death in perihematoma and brain water content could be reduced if argatroban was given intraperitoneally 3 h and 12 h after ICH without adverse affects such as rebleeding.