Affiliation:
1. General Department, Nantong Third People's Hospital Affiliated Nantong Hospital 3 of Nantong University Nantong Jiangsu China
Abstract
AbstractIschemic stroke is a life‐threatening brain disease with the leading cause of disability and mortality worldwide. Heat‐shock protein A12A (HSPA12A) is recognized as a neuroprotective target for treating ischemic stroke; however, its regulatory mechanism has been not fully elucidated yet. Human brain microvascular endothelial cells (hBMECs) were induced by oxygen‐glucose deprivation/reoxygenation (OGD/R) to mimic ischemic stroke. Gain‐ and loss‐of‐function experiments were conducted to explore the regulation of HSAPA12 and PGC‐1α. Cell viability, apoptosis, and permeability were assessed by CCK‐8, TUNEL, and transendothelial electrical resistance (TEER) assays, respectively. The expression of HSPA12A and corresponding proteins was measured by western blot. Cell immunofluorescence was adopted to evaluate ZO‐1 expression. THP‐1 cells were applied to adhere hBMECs in vitro to simulate leukocyte adhesion in the brain. HSPA12A was downregulated in OGD/R‐treated hBMECs. HSPA12A overexpression significantly suppressed OGD/R‐induced cell viability loss and apoptosis in hBMECs. Meanwhile, HSPA12A overexpression attenuated blood–brain barrier (BBB) integrity in OGD/R‐induced hBMECs, evidenced by the restored TEER value and the upregulated ZO‐1, occludin, and claudin‐5. HSPA12A also restricted OGD/R‐induced attachment of THP‐1 cells to hBMECs, accompanied with downregulating ICAM‐1 and VCAM‐1. Additionally, OGD/R‐caused downregulation of PGC‐1α/SIRT3 in hBMECs was partly restored by HSPA12A overexpression. Furthermore, the above effects of HSPA12A on OGD/R‐induced hBMECs injury were partly reversed by PGC‐1α knockdown. HSPA12A plays a protective role against OGD/R‐induced hBMECs injury by upregulating PGC‐1α, providing a potential neuroprotective role of HSPA12A in ischemic stroke.