ZFHX3 silencing alleviates ischemic stroke by suppressing pericyte contraction and calcium influx through inhibiting WNK3 expression.

Zinc finger homeobox 3 (ZFHX3), a zinc finger homeodomain transcription factor, has been implicated in various brain disorders. However, its molecular mechanism in ischemic stroke remains unknown. In this study, we employed a transient middle cerebral artery occlusion/reperfusion (tMCAO/R) model in C57BL/6 mice and subjected mouse primary pericytes to oxygen-glucose deprivation/reoxygenation (OGD/R) treatment. The results revealed a significant upregulation of ZFHX3 expression in tMCAO/R mice in vivo. ZFHX3 knockdown exacerbated ischemic stroke outcomes, as evidenced by increased cerebral infarction volume, elevated risk of brain hemorrhage, aggravated development of cerebral edema, and worsened blood-brain barrier damage in mice with tMCAO/R. Furthermore, ZFHX3 knockdown inhibited calcium influx thereby attenuating pericyte contraction in mice tMCAO/R model. In vitro studies demonstrated that ZFHX3 knockdown significantly upregulated WNK lysine deficient protein kinase 3 (WNK3) expression in OGD/R-treated pericytes. Conversely, WNK3 knockdown relieved ZFHX3 inhibition-induced pericyte contraction in OGD/R-treated pericytes. In conclusion, our study demonstrates that ZFHX3 silencing may confer neuroprotection in ischemic stroke by suppressing pericyte contraction and calcium influx through inhibition of WNK3 expression. These findings position ZFHX3 as a potential therapeutic candidate for the treatment of ischemic stroke.