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背景:先前实验中已经证实补肾活血化痰中药健脑安可抑制脑缺血再灌注后2～24h内糖皮质激素含量增高,降低高糖皮质激素的促神经细胞凋亡毒性作用。这一作用是否在受体环节也有影响尚不清楚。目的:观察中药健脑安对糖皮质激素受体的干预作用,进一步明确健脑安保护脑缺血再灌注后海马神经细胞的作用靶向;并与复方阿米三嗪进行阳性对照。设计:随机对照动物实验。单位:北京中医药大学中心实验室。材料:实验于2002-07/2003-03在北京中医药大学中心实验室进行。取SD雄性大鼠80只,随机分为假手术组、模型组、治疗组、阳性对照组和拮抗剂组5组,每组16只,每组又分为再灌注2,6,12和24h4个时点,每个时点4只。方法:①给药:除模型组外,其他4组大鼠于造模前7d开始灌胃给药,1次/d。假手术组给予7μL/(g·d)蒸馏水;阳性对照组予复方阿米三嗪7.39mg/(kg·d);治疗组予健脑安(肉苁蓉、石菖蒲、大黄等组成)生药6.7g/(kg·d);拮抗剂组给予糖皮质激素受体拮抗剂米非司酮10g/(kg·d),7μL/g。②给药7d后用大脑中动脉阻塞线栓法制备大鼠脑缺血再灌注模型,假手术组麻醉后,剥离颈总动脉后缝合,不予进栓线。主要观察指标:各组在再灌注相应时间点处死大鼠取脑,利用免疫组化方法观察大鼠糖皮质激素受体蛋白表达的变化,计算CA2区3个200倍视野内阳性细胞数。结果:80只大鼠全部进入结果分析。假手术组糖皮质激素受体表达损伤侧与健侧双侧对称,糖皮质激素受体表达无明显差别。模型组、治疗组、阳性对照组、拮抗剂组损伤侧较健侧糖皮质激素受体表达明显减少,亦明显少于假手术组(P<0.05)。治疗组、阳性对照组、拮抗剂组在再灌注各时间点的糖皮质激素受体表达量无明显变化,各组与模型组相比差异不显著(P>0.05)。结论:健脑安对糖皮质激素受体和糖皮质激素含量的调节有选择性,不能调节糖皮质激素受体的表达,与复方阿米三嗪相似;但健脑安可通过降低血浆和脑组织中糖皮质激素含量,发挥对脑缺血再灌注后的神经细胞的保护作用。
BACKGROUND: In previous experiments, it has been confirmed that Bushenhuoxuejing, a traditional Chinese medicine, Jiannaoan can inhibit the increase of glucocorticoid content within 2 to 24 hours after cerebral ischemia and reperfusion, and reduce the neurotoxic apoptosis induced by high glucocorticoid. It is unclear whether this effect has an effect on the receptor link. OBJECTIVE: To observe the intervention effect of JianNaoAn on glucocorticoid receptor and further clarify the effect of Jiannaoan on protection of hippocampal neuronal cells after cerebral ischemia-reperfusion, and to make a positive comparison with compound amifatizine. Design: Randomized controlled animal experiments. Unit: Central Laboratory, Beijing University of Chinese Medicine. MATERIALS: The experiment was performed at the Central Laboratory of Beijing University of Traditional Chinese Medicine from July 2002 to March 2003. Eighty SD male rats were randomly divided into sham operation group, model group, treatment group, positive control group and antagonist group 5 groups, each group was divided into 16 groups. Each group was divided into reperfusion 2, 6, 12 and 24h. At the time, 4 at each time. Methods: 1 Dosing: In addition to the model group, the other four groups of rats were given intragastric administration 7 days before modeling, once/d. 7μL/(g·d) distilled water was given to the sham operation group; 7.90mg/(kg·d) of the compound amifolazine was given to the positive control group; 6.7g of the drug was given to the Jiannaoan (Citrusion, Shichangpu, rhubarb, etc.) (/kg·d); Antagonist group received glucocorticoid receptor antagonist mifepristone 10g/(kg·d), 7μL/g. 2 After 7 days administration, cerebral ischemia-reperfusion model was established by occlusion of middle cerebral artery occlusion. After sham operation group was anaesthetized, common carotid artery was dissected and then sutured. MAIN OUTCOME MEASURES: Each group was sacrificed at the corresponding time point of reperfusion to obtain brains. The changes of glucocorticoid receptor protein expression in rats were observed by immunohistochemical method. The number of positive cells in three 200-fold fields in CA2 region was calculated. Results: All 80 rats entered the results analysis. In the sham operation group, the glucocorticoid receptor expression was bilaterally symmetrical on the injured side and the healthy side, and there was no significant difference in the expression of the glucocorticoid receptor. The expression of the glucocorticoid receptor in the injured side of the model group, the treatment group, the positive control group, and the antagonist group was significantly decreased, and was also significantly lower than that in the sham operation group (P<0.05). The expression of glucocorticoid receptors in the treatment group, positive control group, and antagonist group at each time point did not change significantly, and there was no significant difference between the groups and the model group (P>0.05). Conclusion: Jiannaoan is selective in the regulation of glucocorticoid receptor and glucocorticoid content, and it can not regulate the expression of glucocorticoid receptor, similar to compound amig triazine. However, Jiannaoan can reduce plasma and brain. The content of glucocorticoid in the tissue plays a protective role in nerve cells after cerebral ischemia-reperfusion.