Mechanistic study of butylphthalide in alleviating cerebral Edema after intracerebral Hemorrhage by regulating miR-7-5p expression

Objective: To investigate the mechanisms by which butylphthalide (NBP) alleviates cerebral edema after intracerebral hemorrhage (ICH) through the regulation of miR-7-5p expression.

Methods: 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).

Results: 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.

Conclusion: 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.

Butylphthalide (NBP); Cerebral edema; Intracerebral hemorrhage (ICH); MiR-7-5p; STAT3.