Chen Xiqian, Wang Shige, Lei Qiang, Liu Jia, Lu Wei
Department of Neurology, Second Xiangya Hospital, Central South University, Changsha, China.
Clinical Medical Research Center for Stroke Prevention and Treatment of Hunan Province, Department of Neurology, Second Xiangya Hospital, Central South University, No. 139, Renmin Middle Road, Changsha, Hunan, China.
Metab Brain Dis. 2025 Jul 1;40(6):238. doi: 10.1007/s11011-025-01659-x.
To investigate the mechanisms by which butylphthalide (NBP) alleviates cerebral edema after intracerebral hemorrhage (ICH) through the regulation of miR-7-5p expression.
An ICH model was generated in CTX-TNA2 rat astrocyte cell lines using hemin intervention. An in vivo ICH model was created by injecting type IV collagenase into the basal ganglia of Sprague-Dawley (SD) rats. The cell/rat models were treated with NBP and/or stattic. The expression of STAT3, miR-7-5p, EGFR, PI3K, AKT, p-AKT, AKT2, AKT3, and AQP4 were assessed using qRT-PCR, Western blotting, and immunofluorescence. Brain water content was measured using the wet-to-dry weight method, and neurological deficits were evaluated using the NSS (neurological severity score).
In both the CTX-TNA2 ICH model and the rat ICH model, miR-7-5p expression was significantly reduced, while STAT3, EGFR, AKT, p-AKT, AKT2, AKT3, and AQP4 expression were elevated compared to the blank/sham-operated group. NBP increased the expression of STAT3 and miR-7-5p, while reducing the expression of EGFR, AKT, p-AKT, AKT2, AKT3, and AQP4. NBP also decreased brain water content and improved NSS scores. STAT3 inhibition significantly reduced STAT3 and miR-7-5p expression, increased the expression of EGFR, PI3K, AKT, p-AKT, AKT2, AKT3, and AQP4, and elevated brain water content. NBP can reverse the downregulation of STAT3 and miR-7-5p expression, the upregulation of EGFR/PI3K/AKT axis and AQP4 expression, and the increase in brain water content induced by STAT3 inhibition.
NBP alleviates cerebral edema after ICH by upregulating STAT3 expression, thereby increasing miR-7-5p levels and inhibiting the EGFR/PI3K/AKT axis and AQP4 expression.
探讨丁苯酞(NBP)通过调节miR-7-5p表达减轻脑出血(ICH)后脑水肿的机制。
采用血红素干预在CTX-TNA2大鼠星形胶质细胞系中建立ICH模型。通过向Sprague-Dawley(SD)大鼠基底神经节注射IV型胶原酶建立体内ICH模型。细胞/大鼠模型用NBP和/或Stattic处理。采用qRT-PCR、Western印迹和免疫荧光法评估STAT3、miR-7-5p、表皮生长因子受体(EGFR)、磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(AKT)、磷酸化AKT(p-AKT)、AKT2、AKT3和水通道蛋白4(AQP4)的表达。采用干湿重法测量脑含水量,并用神经功能缺损评分(NSS)评估神经功能缺损。
在CTX-TNA2 ICH模型和大鼠ICH模型中,与空白/假手术组相比,miR-7-5p表达均显著降低,而STAT3、EGFR、AKT、p-AKT、AKT2、AKT3和AQP4表达均升高。NBP增加STAT3和miR-7-5p的表达,同时降低EGFR、AKT、p-AKT、AKT2、AKT3和AQP4的表达。NBP还降低了脑含水量并改善了NSS评分。抑制STAT3可显著降低STAT3和miR-7-5p的表达,增加EGFR、PI3K、AKT、p-AKT、AKT2、AKT3和AQP4的表达,并升高脑含水量。NBP可逆转由抑制STAT3所诱导的STAT3和miR-7-5p表达下调、EGFR/PI3K/AKT轴和AQP4表达上调以及脑含水量增加。
NBP通过上调STAT3表达,从而增加miR-7-5p水平并抑制EGFR/PI3K/AKT轴和AQP4表达,减轻ICH后脑水肿。