| 刘甜甜；马春雷；刘付红；郭平.化痰通络汤改善脑缺血再灌注小鼠神经功能作用的研究[J].中医药信息,2023,40(11):23-31 |
| 化痰通络汤改善脑缺血再灌注小鼠神经功能作用的研究 |
| Research on the Effect of Huatan Tongluo Decoction on Improving Neural Function of Cerebral Ischemia－Reperfusion Mice |
| 投稿时间：2023-03-15 录用日期：2023-04-10 |
| DOI：10.19656/j.cnki.1002-2406.20231104 |
| 中文关键词: 脑缺血再灌注 化痰通络汤 血管性血友病因子 血小板 血脑屏障 |
| 英文关键词: Cerebral ischemia reperfusion Huatan Tongluo Decoction Von Willebrand factor Platelets Blood－brain barrier |
| 基金项目:双氢青蒿素对血管炎（湿热伤脉型）的保护作用及其机制研究 |
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| 中文摘要: |
| 目的：探究化痰通络汤对大脑中动脉缺血再灌注（MCAO/R）模型小鼠神经功能的作用及分子机制。方法：85只SPF级C57BL/6J雄性小鼠随机分为假手术组（Control）、模型组（Model）、化痰通络汤高剂量组（HTTLD-H）、化痰通络汤中剂量组（HTTLD-M）、化痰通络汤低剂量组（HTTLD-L）和阿司匹林肠溶片组（AECT）。小鼠MCAO/R模型复制成功30 min后，分别给予相应药物灌胃给药，连续干预7 d。采用神经功能缺损评分评估小鼠神经功能缺损程度；2,3,5-氯化三苯基四氮唑（TTC）染色法检测脑组织梗死体积；苏木素/伊红（HE）染色法检测脑组织形态学改变；免疫蛋白印记（Western blot）检测脑组织中血管性血友病因子（vWF）、纤维蛋白原（Fibrinogen）、血脑屏障密封蛋白（Claudin-5）和闭合蛋白（Occludin）蛋白表达水平；免疫荧光法检测脑组织中vWF、血小板膜糖蛋白Ⅸ（GPⅨ，标记血小板）、Fibrinogen、Claudin-5和Occludin表达。结果：与模型组比较，化痰通络汤能减少MCAO/R小鼠脑组织梗死体积（P <0.01），改善神经功能（P <0.01），以化痰通络汤中剂量组效果最佳；化痰通络汤能在一定程度上维持MCAO/R小鼠脑组织中血管壁完整性，改善血液循环；化痰通络汤能减少脑组织中vWF表达（P <0.01）；减少血小板黏附和聚集（P <0.01）；减少Fibrinogen沉积，以化痰通络汤高、中剂量组效果最显著（P <0.01）；化痰通络汤能升高脑组织中Claudin-5和Occludin蛋白表达（P <0.05），维护血脑屏障。结论：化痰通络汤能减少MCAO/R模型小鼠脑组织梗死体积，改善神经功能，这可能与化痰通络汤降低MCAO/R模型小鼠脑组织中vWF的表达，减少血小板黏附和聚集，减少Fibrinogen沉积，并在一定程度上维持Claudin-5和Occludin蛋白表达、维护血脑屏障有关。 |
| 英文摘要: |
| Objective: To explore the effects and molecular mechanism of Huatan Tongluo Decoction on the neural function of middle cerebral artery ischemia and reperfusion(MCAI/R) model mice. Method: 85 SPF C57BL/6J male mice were randomly divided into sham operation group(Control), model group(Model), Huatan Tongluo Decoction high-dose group(HTTLD-H), Huatan Tongluo Decoction medium-dose group(HTTLD-M), Huatan Tongluo Decoction low-dose group(HTTLD-L), and aspirin enteric-coated tablet group(AECT). 30 min after successful replication of MCAI/R model mice, corresponding drugs were administered to the mice by gavage for 7 consecutive days. Neurological impairment scores were used to evaluate the degree of neurological impairment of the mice. The infarct volume of brain tissue was detected by 2,3,5-triphenyltetrazolium chloride(TTC) staining. Hematoxylin-eosin(HE) staining was used to detect morphological changes in the brain tissue. Western blot was used to detect the expression levels of Von Willebrand factor(v WF), fibrinogen, claudin-5 and occludin of the brain tissue; the expression of v WF, platelet membrane glycoprotein Ⅸ （GP Ⅸ, labeled platelets）, fibrinogen, claudin-5 and occludin in the brain tissue were detected by immunofluorescence. Results: Compared with the treatment of model group, Huatan Tongluo Decoction can reduce the infarct volume of brain tissue in MCAI/R mice(P < 0. 01) and improve the neurological function(P < 0. 01), and the medium dose group had the best effect. The decoction can maintain the integrity of blood vessel wall and improve blood circulation in brain tissue of MCAI/R mice to a certain extent. It can decrease the expression of v WF in brain tissue(P < 0. 01) and reduce platelet adhesion and aggregation(P < 0. 01), and the decrease of fibrinogen deposition has the most significant difference between the high dose and medium dose(P < 0. 01). The decoction can increase the expression of claudin-5(P <0. 05) and occludin(P < 0. 05) in brain tissue to maintain the blood-brain barrier. Conclusion: Huatan Tongluo Decoction can reduce cerebral infarction volume and improve nerve function in MCAI/R model mice, which may be related to the fact that Huatan Tongluo Decoction reduces vWF expression in brain tissue of MCAI/R model mice, reduces platelet adhesion and aggregation, and reduces fibrinogen deposition. To a certain extent, the decoction can maintain claudin-5 and occludin expressions, and maintain blood-brain barrier. |
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