目的 探讨脑泰方通过miRNA-124/信号转导和转录激活因子3（signal transducer and activator of transcription 3，STAT3）信号通路减轻脑缺血再灌注损伤（cerebral ischemia-reperfusion injury，CIRI）后胶质瘢痕的作用。方法 将48只SD大鼠随机分为假手术组、模型组、脑泰方、脑泰方＋miRNA-124拮抗剂组，每组12只。除假手术组外，各组采用大脑中动脉阻塞/复灌注方法建立大鼠CIRI模型。脑泰方＋miRNA-124拮抗剂组于术前20 min侧脑室注射miR-124-3P antagomir，给予脑泰方连续治疗14 d后，采用Longa法进行神经功能评分；水迷宫实验检测大鼠学习和记忆能力；旷场实验评估大鼠焦虑程度；TTC染色法检测脑梗死面积；苏木素-伊红（HE）染色法观察脑组织病理变化；免疫荧光法检测缺血脑皮质胶质纤维酸性蛋白（glial fibrillary acidic protein，GFAP）的表达；qRT-PCR检测脑缺血皮质组织miRNA-124基因表达；Western blotting检测脑缺血皮质组织M2型小胶质细胞标志分子[精氨酸酶-1（arginase-1，Arg-1）、CD206]，炎症因子[白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、IL-4]，胶质瘢痕相关蛋白[GFAP、神经蛋白聚糖（neurocan）、磷酸聚糖（phosphacan）]以及p-STAT3、STAT3蛋白表达。结果 与模型组比较，脑泰方组及脑泰方＋miRNA-124拮抗剂组大鼠脑组织病理损伤减轻，脑梗死面积减少（P＜0.05、0.01）；水迷宫和旷场实验结果显示，各给药组大鼠认知功能改善，卒中后焦虑减轻（P＜0.05、0.01）；免疫荧光结果显示，各给药组大鼠脑缺血皮质组织GFAP荧光表达减少（P＜0.01），瘢痕厚度减轻；qRT-PCR结果显示，各给药组大鼠脑缺血皮质组织miRNA-124基因表达水平升高（P＜0.01）；Western blotting结果显示，各给药组大鼠脑缺血皮质组织GFAP、neurocan、phosphacan、p-STAT/STAT3、IL-1β、IL-6蛋白表达水平显著降低（P＜0.05、0.01），CD206、Arg-1、IL-4蛋白表达水平显著升高（P＜0.05、0.01）。结论 脑泰方对CIRI具有神经保护作用，可能与通过miRNA-124/STAT3信号通路减轻CIRI大鼠星形胶质细胞增生和神经胶质瘢痕形成有关。

Objective To investigate the effect of Naotaifang (脑泰方, NTF) on reducing glial scar formation after cerebral ischemia-reperfusion injury (CIRI) through miRNA-124/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Methods A total of 48 SD rats were randomly divided into sham group, model group, NTF group and NTF + miRNA-124 antagonist group, with 12 rats in each group. Except for sham group, CIRI model was established by middle cerebral artery occlusion/reperfusion in each group. NTF + miRNA-124 antagonist group was injected with miR-124-3P antagomir into the lateral ventricle 20 min before surgery. After continuous treatment with NTF for 14 d, the neural function score was evaluated using Longa method; The learning and memory ability of rats was detected by the water maze experiment. The anxiety degree of rats was evaluated by open field experiment. The infarct size was detected by TTC staining. The pathological changes of brain were observed by hematoxylin-eosin (HE) staining. The expression of glial fibrillary acidic protein (GFAP) in ischemic cerebral cortex was detected by immunofluorescence. The expression of miRNA-124 gene in cerebral ischemic cortex was detected by qRT-PCR. Western blotting was used to detect M2 microglial cell marker molecules [arginase-1 (Arg-1), CD206], inflammatory factors [interleukin-1β (IL-1β), IL-6, IL-4], glial scar associated proteins [GFAP, neurocan, phosphatan], as well as p-STAT3 and STAT3 protein expressions in cerebral ischemic cortex.Results Compared with model group, the pathological injury of brain tissue in NTF group and NTF + miRNA-124 antagonist group was reduced, and the cerebral infarction area was decreased (P < 0.05, 0.01). The results of water maze and open field experiments showed that the cognitive function of rats in each administration group were improved, and the anxiety after stroke was alleviated (P < 0.05, 0.01). The results of immunofluorescence showed that the fluorescence expression of GFAP was decreased and scar thickness was decreased (P < 0.01). The results of qRT-PCR showed that the expression of miRNA-124 gene in cerebral ischemic cortex tissues of rats in each administration group was increased (P < 0.01). The results of Western blotting showed that the expressions of GFAP, neurocan, phosphacan, p-STAT/STAT3, IL-1β and IL-6 protein in cerebral ischemic cortical tissue of rats in each administration group were significantly decreased (P < 0.05, 0.01), the expressions of CD206, Arg-1 and IL-4 were significantly increased (P < 0.05, 0.01). Conclusion NTF exhibits a neuroprotective effect on CIRI, which may be attributed to the reduction of astrocyte proliferation and glial scar formation in CIRI rats through miRNA-124/STAT3 signaling pathway.

R285.5

国家自然科学基金项目（82174167）；湖南省自然科学基金面上项目（2023JJ30464）；湖南省教育厅青年项目（22B0382）；湖南省卫生健康委科研项目（D202303078170）