目的 探討腦泰方通過miRNA-124/信號轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子3（signal transducer and activator of transcription 3，STAT3）信號通路減輕腦缺血再灌注損傷（cerebral ischemia-reperfusion injury，CIRI）后膠質(zhì)瘢痕的作用。方法 將48只SD大鼠隨機分為假手術(shù)組、模型組、腦泰方、腦泰方＋miRNA-124拮抗劑組，每組12只。除假手術(shù)組外，各組采用大腦中動脈阻塞/復(fù)灌注方法建立大鼠CIRI模型。腦泰方＋miRNA-124拮抗劑組于術(shù)前20 min側(cè)腦室注射miR-124-3P antagomir，給予腦泰方連續(xù)治療14 d后，采用Longa法進行神經(jīng)功能評分；水迷宮實驗檢測大鼠學(xué)習(xí)和記憶能力；曠場實驗評估大鼠焦慮程度；TTC染色法檢測腦梗死面積；蘇木素-伊紅（HE）染色法觀察腦組織病理變化；免疫熒光法檢測缺血腦皮質(zhì)膠質(zhì)纖維酸性蛋白（glial fibrillary acidic protein，GFAP）的表達；qRT-PCR檢測腦缺血皮質(zhì)組織miRNA-124基因表達；Western blotting檢測腦缺血皮質(zhì)組織M2型小膠質(zhì)細(xì)胞標(biāo)志分子[精氨酸酶-1（arginase-1，Arg-1）、CD206]，炎癥因子[白細(xì)胞介素-1β（interleukin-1β，IL-1β）、IL-6、IL-4]，膠質(zhì)瘢痕相關(guān)蛋白[GFAP、神經(jīng)蛋白聚糖（neurocan）、磷酸聚糖（phosphacan）]以及p-STAT3、STAT3蛋白表達。結(jié)果 與模型組比較，腦泰方組及腦泰方＋miRNA-124拮抗劑組大鼠腦組織病理損傷減輕，腦梗死面積減少（P＜0.05、0.01）；水迷宮和曠場實驗結(jié)果顯示，各給藥組大鼠認(rèn)知功能改善，卒中后焦慮減輕（P＜0.05、0.01）；免疫熒光結(jié)果顯示，各給藥組大鼠腦缺血皮質(zhì)組織GFAP熒光表達減少（P＜0.01），瘢痕厚度減輕；qRT-PCR結(jié)果顯示，各給藥組大鼠腦缺血皮質(zhì)組織miRNA-124基因表達水平升高（P＜0.01）；Western blotting結(jié)果顯示，各給藥組大鼠腦缺血皮質(zhì)組織GFAP、neurocan、phosphacan、p-STAT/STAT3、IL-1β、IL-6蛋白表達水平顯著降低（P＜0.05、0.01），CD206、Arg-1、IL-4蛋白表達水平顯著升高（P＜0.05、0.01）。結(jié)論 腦泰方對CIRI具有神經(jīng)保護作用，可能與通過miRNA-124/STAT3信號通路減輕CIRI大鼠星形膠質(zhì)細(xì)胞增生和神經(jīng)膠質(zhì)瘢痕形成有關(guān)。

Objective To investigate the effect of Naotaifang (腦泰方, 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

國家自然科學(xué)基金項目（82174167）；湖南省自然科學(xué)基金面上項目（2023JJ30464）；湖南省教育廳青年項目（22B0382）；湖南省衛(wèi)生健康委科研項目（D202303078170）