CD147 exacerbates ischemic brain injury and disrupts Wnt/β-catenin signaling and blood-brain barrier integrity

Ischemic stroke is a leading cause of death and disability, with limited therapies beyond reperfusion. CD147 (Basigin), upregulated after cerebral ischemia, participates in neuroinflammation, however, its precise contributions to blood-brain barrier (BBB) disruption and neuronal injury are incompletely understood. Using a photothrombotic permanent middle cerebral artery occlusion (pMCAO) model in C57BL/6J male adult mice, we delivered CD147-overexpressing adenovirus (Ad Bsg) or control fluorophore adenovirus (Ad Fc) via intracerebroventricular injection.

Infarct volume, neurological deficits, motor function, neuronal apoptosis, BBB integrity and hemorrhagic transformation were assessed. In vitro, HT22 neuronal cells were treated with recombinant Cyclophilin A (CyPA) to activate CD147 signaling. Wnt/β-catenin pathway activity was evaluated by western blot and qPCR.

Using constitutively active β-catenin mice and pharmacological activation of AKT (using SC-79) were also performed. CD147 expression peaked at 24 h after stroke and correlated with infarct volume and neurological deficit severity. CD147 overexpression significantly increased infarct size, neuronal apoptosis, BBB leakage, brain edema, and downregulated the expression of claudin-5 and ZO-1.

In neurons, CyPA-induced CD147 activation reduced p-AKT, p-GSK3β, β-catenin, and Wnt target gene expression. These effects were reversed by CD147 knockdown or by constitutively active β-catenin. CD147 exacerbates ischemic brain injury and BBB disruption by suppressing neuronal Wnt/β-catenin signaling through a mechanism involving the AKT/GSK3β node, supporting further evaluation of CD147-directed interventions after stroke.

Cluster of differentiation 147 (Basigin) Glyceraldehyde‑3‑phosphate dehydrogenase Immortalized murine hippocampal neuronal cell line Institutional Animal Care and Use Committee Platelet‑derived growth factor receptor beta Sodium dodecyl sulfate–polyacrylamide gel electrophoresis Terminal deoxynucleotidyl transferase dUTP nick end labeling We are grateful for the technical support from the Animal Facilities of Shenzhen Institutes of Advanced Technology (SIAT), the Chinese Academy of Sciences (CAS) and for the support from UCAS Scholarship for Young Talents. This work was supported by the National Natural Science Foundation of China (31900704 to C.F., 82204983 to Z.M., 82273923 to Y.M.), National Key Research and Development Program of China (2023YFE0202200), Guangdong Basic and Applied Basic Research Foundation (2023A1515010467 to C.F., 2021Β1515120089 to Y.M.), Shenzhen Science and Technology Program (JCYJ20230807140721043 to C.F., JCYJ20220531092010023, JCYJ20240813152424031 to Z.M.), International Cooperation Project of the Shenzhen Municipal Science and Technology Innovation Commission (GJHZ20240218112559009), Shenzhen Medical Research Fund (D2403002), Sanming Project of Medicine in Shenzhen (No. SZZYSM202411012 to Z.M.).

The funders were not involved in the study design, data collection, data analysis, manuscript preparation or publication decisions. Yueqing Wang, Femi Abiola Ogunleye and Chonglin Xu contributed equally to this work. State Key Laboratory of Biomedical Imaging Sciences and Systems, Guangdong-Hong Kong Joint Laboratory for Metabolic Medicine, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong, China Yueqing Wang, Femi Abiola Ogunleye, Chonglin Xu, Yumei Zhou, Yinzhong Ma & Cheng Fang University of Chinese Academy of Sciences, Beijing, China Yueqing Wang, Femi Abiola Ogunleye & Yumei Zhou Department of Biomedical Engineering, Southern University of Science and Technology, Shenzhen, Guangdong, PR China Emergency Department, Shenzhen Traditional Chinese Medicine Hospital, The Fourth Clinical College of Guangzhou University of Chinese Medicine, Shenzhen, Guangdong, China Institute of Biomedicine and Biotechnology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Xueyuan Avenue 1068, Nanshan, Shenzhen, 518055, Guangdong, China Shenzhen Traditional Chinese Medicine Hospital, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, No.1 Fuhua Road, Futian District, Shenzhen, 518033, Guangdong, China Correspondence to Yinzhong Ma, Zhizhun Mo or Cheng Fang.