Elucidation of the mechanism of rho-kinase induced apoptotic neuronal cell death

SD大鼠采用腔内缝合技术制作大脑中动脉闭塞模型，持续90分钟。大鼠再灌流24小时后断头进行脑梗塞和脑水肿分析。实验动物随机分为以下四组。(1)赋形剂对照组：小剂量法舒地尔(3 mg/kg)治疗+常温(37℃)组(3)法舒地尔(3 mg/kg)治疗+低温(35℃)组(4)依达拉奉治疗组。Rho-Kinase抑制剂处理组动物在缺血60min后持续静脉注射法舒地尔(3.0 mg/kg)，而赋形剂治疗组给予相同剂量的赋形剂。在缺血期间，监测实验动物的颞肌和直肠温度，并将其维持在37或35度。神经症状评估(体位和偏瘫)在脑梗塞和水肿分析前立即进行。此外，在缺血和再灌流过程中，测量大脑皮质血流量。Physii…多项生理参数(PH、P02、二氧化碳分压、血糖、脑温、体温)组间无统计学差异。皮质血流量在组间也无统计学差异。低剂量法舒地尔(3.0 mg/kg)与车辆加热的动物相比，婴儿体积更小。此外，与37℃法舒地尔治疗的动物相比，超亚低温(35℃)法舒地尔组的心肌梗死面积更小。低剂量法舒地尔治疗组(III组)皮质和纹状体梗死体积(85±42mm3)明显小于I组(195±36mm3，P<0.001)。常规剂量法舒地尔+亚低温组(IV组)皮质或纹状体梗死体积(76±49 mm~3，35±22 mm~3)较I组(195±36 mm~3，83±20 mm~3)显著缩小(P&lt；0.001，P&lt；0.01)，而单纯低温组(II组)未见明显缩小。与I组(73±25 mm~3)相比，低剂量法舒地尔(IV组)并不能显著减少纹状体水肿体积(38±14 mm~3)，但与I组(35±22 mm~3)相比，低温组(IV组)皮质脑梗塞体积(35±22 mm~3)明显缩小(p&lt；0.001)，可使半影区促凋亡bax蛋白表达减少，抗凋亡蛋白Bc l-2上调。较少

Sprague-Dawley rats were subjected to MCAo using an intraluminal suture technique fir 90 minutes. The rats were reperfused for 24 hours and decapitated for infarct and edema analysis . Animals were randomly divided into the following four groups. (I) vehicle-treated control group 0:) low dose of fasudil (3.0 mg/kg)-treated+ normothennia (37℃) group (III) fasudil (3.0 mg/kg)treated + hypothermia (35℃) group (IV) edaravone-treated group. A rho-kinase inhibitor-treated animals received a continuous injection of fasudil (3.0 mg/kg) intravenously fir 60 minutes after the onset of ischemia, while vehicle-treated groups received same dose of vehicle. During ischemia, temporal muscle and rectal temperatures were monitored and maintained at 37 or 35 degrees in the experimental animals. Neurological symptom evaluations (posture and hemiplegia) were perfumed immediately before infarct and edema analysis. Furthermore, cortical cerebral blood flow was measured during ischemia and reperfusion. Physi … More ological Paramers (PH, P02, PCO2, Blood Sugar, Brain and Body Temperature) has no statistical difference between the groups. The cortical blood flow also has no statistical difference between the groups. Low dose fasudil (3.0 mg/kg) showed infant volume smaller compared to the vehicle-heated animal. Furthermore, extra-mild hypothermia (35℃) with fasudil showed infarct more smaller compared to 37℃ fasudil treated animal. The cortical and striatal infarct volume (85±42 mm3) in the low dose of fasudil-treated group (group III) was significantly less than those in group I (195±36 mm3, p<0.001). Law dose Fasudil + hypothermia (group IV) decreased the cortical or striatal infarct volume ( 76±49 mm3, 35±22 mm3) significantly (p<0.001, P<0.01, respectively) compared with those of groups I (195 ± 36 mm3, 83 ± 20 mm3 ), whereas hypothermia-treated only (group II) did not decreased cortical and striatal infarct volume significantlyLow dose Fasudil (group II) decreased the cortical edema volume (34±18 mm3,) significantly (p<0.05) compared with those of groups I (73 ± 25 mm3), whereas low dose fasudil did not decreased the striatal edema volume (38±14 mm3) significantly Fasudil + hypothermia (group IV) decreased the cortical infarct volume ( 35 ± 22 mm3) significantly (p<0.001) compared with those of groups I Fasudil reduced expression of the proapoptotic Bax protein in the penumbra, whereas it inc eased upregulation of the antiapoptotic protein Bcl-2 in this area Furthermore, TUNEL positive rats were decreased after administration of fasudil in the penumbra. Less