雌激素、灯盏花对去势大鼠脑缺血损伤保护作用的叠加实验

Separate and combined neuroprotection of estrogen and breviscapini against cerebral ischemic injury of ovariectomized rats

目的:探讨去势大鼠局灶脑缺血再灌注后雌激素、灯盏花的叠加保护作用。方法:实验于1998-06/1999-04在华西医科大学的相关实验室进行。雌性SD大鼠进行双侧卵巢切除后2周再采用线栓法制备去势大鼠大脑中动脉缺血(2h)再灌注(70h)模型。50只SD大鼠随机分为雌激素组,灯盏花组、雌激素加灯盏花组、对照组和假手术组。再灌注2h和70h后分别进行神经功能缺损评分。再灌注70h显微镜下观察脑损伤区组织病理学、脑梗死体积比和脑水肿体积,测定血液中一氧化氮、白细胞介素1(IL-1)及肿瘤坏死因子(TNF)水平的变化,采用免疫组织化学方法观察脑内雌激素受体的表达。结果:再灌注70h后,与对照组神经功能缺损评分犤(1.7±0.6)分犦相比,雌激素组犤(0.3±0.5)分犦、灯盏花组犤(0.9±0.6)分犦、雌激素加灯盏花组犤(0.2±0.4)分犦明显降低(F=13.488,P<0.05);与对照组脑梗死体积比犤(31.33±1.26)%犦相比,雌激素组犤(12.52±1.40)%犦、灯盏花组犤(18.35±1.10)%犦、雌激素加灯盏花组犤(11.52±0.69)%犦明显降低(F=612.876,P<0.01);与对照组脑水肿体积犤(69.77±3.40)mm3犦相比,雌激素组犤(32.59±2.12)mm3犦、灯盏花组((51.2±4.37)mm3)、雌激素加灯盏花组犤(30.97±2.13)mm3犦明显降低(F=334.867,P<0.01);与对照组IL-1水平犤(0.

AIM: To study the separate and combined neuroprotection provided by estrogen and breviscapini after cerebral ischemic reperfusion in ovariectomized rats.METHODS: The experiment was performed in the Affiliated Laboratories of West China University of Medical Sciences from June 1998 to April 1999. Female SD rats bilaterally ovariectomized two weeks before were subjected to 2- hour middle cerebral artery occlusion (MCAO) with an intraluminal filament, followed by a reperfusion of 70 hours. Fifty SD rats were randomized divided into 5 groups: estrogen group, breviscapini group, estrogen plus breviscapini group, control group and sham operation group.The neurologic impairment score of each rat was evaluated after reperfusion of 2 hours and 70 hours. The histopathology in brain injury area, changes of cerebral infarction volume ratio and edema volume were investigated under microscope after reperfusion of 70 hours. The content of nitric oxide (NO) in serum, the changes of interleukin- 1(IL- 1) and tumor necrosis factor(TNF) were measured after reperfusion of 70 hours. The expression of estrogen receptor in brain was investigated by using immunohistochemistry method after reperfusion of 70 hours.RESULTS: After the reperfusion of 70 hours, the neurologic impairment scores in estrogen group (0.3± 0.5), breviscapini group (0.9± 0.6), and estrogen plus breviscapini group(0.2± 0.4) were significantly reduced than that of control group(1.7± 0.6) (F=13.488,P< 0.01). Cerebral infarction volume ratio in in estrogen group [(12.52± 1.40)% ], breviscapini group [(18.35± 1.10)], and estrogen plus breviscapini group[(11.52± 0.69)% ] were also significantly reduced than that of control group[(31.33± 1.26)% ] (F= 612.876, P< 0.01). Cerebral edema volume in estrogen group [(32.59± 2.12) mm3], breviscapini group [(51.2± 4.37) mm3], and estrogen plus breviscapini group [(30.97± 2.13) mm3]were significantly reduced than that of control group [(31.33± 1.26)% ] (F=334.867,P < 0.01).The levels of IL- 1 in estrogen group [(0.84± 0.03) μ g/L] breviscapini group [(0.84± 0.06) μ g/L], and estrogen plus breviscapini group [(0.82± 0.02) μ g/L] were significantly reduced than that of control group [(0.97± 0.08) μ g/L](F=24.626, P < 0.01).The levels of TNF in estrogen group [(150.38± 10.85) μ g/L], breviscapini group [(157.13± 11.08) μ g/L], and estrogen plus breviscapini group [(149.5± 3.95) μ g/L] were also significantly reduced than that of control group [171.25± 3.95) μ g/L] (F=31.75,P< 0.01). The levels of NO in estrogen group [(64.58± 6.55) μ mol/L], breviscapini group [(78.82± 7.33) μ mol/L], and estrogen plus breviscapini group [(62.5± 4.77) μ mol/L] were significantly reduced than that of control group [(133. 41± 8.45) μ mol/L] (F=249.945, P< 0.01). CONCLUSION: Estrogen can provide significantly neuroprotective effect for ovariectomized rat with cerebral ischemia. Estrogen is better than breviscapini. Combination therapy with estrogen plus breviscapini is not superior to separate therapy with estrogen alone.