| 摘要: |
| 目的 观察杜蛭丸对缺血性脑卒中大鼠的脑保护作用并探究其机制。方法 将健康雄性SD大鼠分为5组：假手术组、模型组、杜蛭丸低剂量组（每天1.0 g/kg）、杜蛭丸高剂量组（每天2.0 g/kg）、阿加曲班组（阳性对照）。杜蛭丸低、高剂量组每日灌胃给药1次，其他组灌胃给予等量生理盐水，连续8 d。除假手术组外，其他组大鼠均于第8天灌胃后0.5 h采用线栓法建立大脑中动脉闭塞（MCAO）脑缺血再灌注模型，缺血时间为90 min。阿加曲班组在再灌注同时经尾静脉单次给予阿加曲班（3.0 mg/kg）。脑缺血24 h后，采用2,3,5-氯化三苯基四氮唑（TTC）染色、尼氏染色和神经行为学评分分别对大鼠的脑梗死面积、脑梗死程度和神经功能损伤情况进行评估，采用ELISA法检测脑缺血后脑内炎症因子TGF-β1、IL-1β水平，应用全自动凝血分析仪测定凝血指标。应用2,2-二苯基-1-苦基肼（DPPH）清除实验分析杜蛭丸对自由基的清除能力。结果 高剂量杜蛭丸可减少MCAO大鼠脑梗死面积、改善神经行为缺损症状（与模型组比较，均P＜0.01），并降低神经元尼氏染色评分。模型组大鼠缺血脑组织炎症因子IL-1β、TGF-β1水平升高（与假手术组比较，均P＜0.01），而给予低剂量和高剂量杜蛭丸后大鼠脑组织IL-1β、TGF-β1水平均降低（与模型组比较，均P＜0.01）。模型组大鼠凝血指标纤维蛋白原水平高于假手术组（P＜0.01），低剂量和高剂量杜蛭丸均可抑制纤维蛋白原的产生（与模型组比较，均P＜0.01）。DPPH清除实验结果显示，杜蛭丸具有一定的自由基清除能力，IC50为（1.33±1.11）mg/mL，曲线坡度为1.378±0.145。结论 杜蛭丸对缺血性脑卒中大鼠有脑保护作用，其机制可能与其抑制纤维蛋白原产生、降低脑内炎症因子TGF-β1和IL-1β水平及抗氧化作用有关。 |
| 关键词: 杜蛭丸 缺血性脑卒中 炎症因子 纤维蛋白原 抗氧化 |
| DOI：10.16781/j.CN31-2187/R.20230253 |
| 投稿时间：2023-05-06修订日期：2024-05-07 |
| 基金项目:上海市生物物质成药性评价专业技术服务平台项目（18DZ2290900）. |
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| Cerebral protective effect of Duzhi pills on ischemic stroke rats and its mechanisms |
| WANG Huihao,LIU Qing,XIE Wei,LIU Li*,WANG Zhiyong* |
(National Key Laboratory of Lead Druggability Research, China State Institute of Pharmaceutical Industry, Shanghai 200437, China
*Corresponding authors) |
| Abstract: |
| Objective To investigate the cerebral protective role of Duzhi pills on ischemic stroke rats and its mechanisms. Methods The healthy male SD rats were assigned to 5 groups: sham group, model group, Duzhi pill low-dose group (1.0 g/kg a day), Duzhi pill high-dose group (2.0 g/kg a day), or argatroban group (positive control). The low-dose and high-dose groups of Duzhi pills were intragastrically administered once a day for 8 d, while the other groups were intragastrically administered with the same amount of normal saline. Except the sham group, the middle cerebral artery occlusion (MCAO) cerebral ischemia-reperfusion model was established by suture method at 0.5 h after intragastric administration on the 8th day in other groups, and the ischemia duration was 90 min. Argatroban group was given a single dose (3.0 mg/kg) of argatroban via caudal vein at the same time of reperfusion. After 24 h of cerebral ischemia, the area of cerebral infarction, degree of cerebral infarction and neurological function injury were evaluated by 2,3,5-triphenyltetrazolium chloride (TTC) staining, Nissl staining and neurobehavioral score, respectively. The levels of inflammatory factors (transforming growth factor-β1 ［TGF-β1］ and interleukin-1β ［IL-1β］) in the brain after cerebral ischemia were detected by enzyme-linked immunosorbent assay (ELISA), and the coagulation indexes were detected by an automatic coagulation analyzer. The scavenging ability of Duzhi pills on free radicals was analyzed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging experiment. Results High-dose Duzhi pills reduced the cerebral infarction area, decreased the deep Nissl staining score of neural cells, and improved the symptoms of neurobehavioral defects in MCAO rats (compared with the model group, all P＜0.01). The levels of IL-1β and TGF-β1 in the ischemic brain tissue of the model group were significantly increased (compared with the sham group, both P＜0.01), while the levels of IL-1β and TGF-β1 in the brain tissue of the rats treated with low-dose and high-dose Duzhi pills were significantly decreased (compared with the model group, all P＜0.01). The coagulation index fibrinogen level in the model group was significantly higher than that in the sham group (P＜0.01), and both low-dose and high-dose Duzhi pills could inhibit the production of fibrinogen (compared with the model group, both P＜0.01). DPPH scavenging test results showed that Duzhi pills could scavenge free radical, the half inhibitory concentration was (1.33±1.11) mg/mL, and the slope of the curve was 1.378±0.145. Conclusion Duzhi pills play a cerebral protective role in ischemic stroke rats. The mechanism may be related to the inhibition of fibrinogen production, the reduction of inflammatory factors (TGF-β1 and IL-1β), and the antioxidant effects. |
| Key words: Duzhi pills ischemic stroke inflammatory cytokines fibrinogen antioxidation |
