立体定向脑室注射代谢性谷氨酸受体1拮抗剂对SAH损伤影响的实验

Effect of the metabotropic glutamate receptors1 antagonist on subarachnoid hemorrhage through sterotactic injection into brain ventricle

目的研究侧脑室注射代谢性谷氨酸受体1(mGluR1)拮抗剂LY367385是否能减轻小鼠蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后脑组织损伤。方法采用非开颅血管内线栓法制备小鼠SAH模型,随机分为3组:对照组(control组)、模型组[(SAH+生理盐水(NS)组)]、mGluR1拮抗剂LY367385组(SAH+LY367385组),于SAH后10min侧脑室注射NS或LY367385(500nmol)5μl,术后行神经功能评分。分别在SAH后6、24、48h3个时相点取右侧脑组织标本,采用蛋白免疫印迹法(Western biotting)检测mGluR1、p-ERK1/2蛋白的表达;用原位末端标记法(TUNEL)检测神经细胞凋亡情况。结果与对照组比较,模型组小鼠神经功能评分显著降低,各组小鼠mGluR1和p-ERK1/2蛋白均有不同程度增强,凋亡细胞增多。与模型组比较,拮抗剂组小鼠神经功能评分增加,mGluR1、p-ERK1/2蛋白表达均有不同程度下调,神经细胞凋亡数目减少。结论 (1)SAH后mGluR1的表达增强可通过激活ERK信号途径诱导神经细胞凋亡;(2)mGluR1选择性拮抗剂LY367385对脑血管痉挛(cerebral vaso-spasm,CVS)损伤具有保护作用。

Objective To investigate the influence of metabotropic glutamate receptorsl antagonist into brain ventricle on neuronal damage after subarachnoid hemorrhage in mice. Methods SAH model was made by endovascular perforation without opening cranium. ICR mice were randomly divided into three groups ： control group, NS ＋ SAH group and LY367385 ＋ SAH group. 10 minutes after SAH ,5pL1 of LY367385 or NS were microinjected into lateral cerebral ventricle respectively, and then neurological score was examined. At different time points （6,24,48h） after operation, Western blot assay was used to ex- amine mGluR1 and p-ERK1/2 expression. Apoptotic cell number were determined by TUNEL method. Results In SAH ＋ NS group ,the neurological function scores were significantly lower, the mGluR1, p-ERK1/2 level and the number of apoptosis cells was significantly enhanced. Mice treated with LY367385 had reduction in mGluR1 ,ERK1/2 activity and neuronal apopto- sis. Neurological scores were relieved. Conclusion The study indicates that injecting metabotropic glutamate receptorsl an- tagonist into brain ventricle may reduce the neuronal damage in mice subjected to subarachnoid hemorrhage.