NAD⁺ rescues aging-induced blood-brain barrier damage via the CX43-PARP1 axis

Blood-brain barrier (BBB) function deteriorates during aging, contributing to cognitive impairment and neurodegeneration. It is unclear what drives BBB leakage in aging and how it can be prevented. Using single-nucleus transcriptomics, we identified decreased connexin 43 (CX43) expression in cadherin-5⁺ (Cdh5⁺) cerebral vascular cells in naturally aging mice and confirmed it in human brain samples. Global or Cdh5⁺ cell-specific CX43 deletion in mice exacerbated BBB dysfunction during aging. The CX43-dependent effect was not due to its canonical gap junction function but was associated with reduced NAD⁺ levels and mitochondrial dysfunction through NAD⁺-dependent sirtuin 3 (SIRT3). CX43 interacts with and negatively regulates poly(ADP-ribose) polymerase 1 (PARP1). Pharmacologic inhibition of PARP1 by olaparib or nicotinamide mononucleotide (NMN) supplementation rescued NAD⁺ levels and alleviated aging-associated BBB leakage. These findings establish the endothelial CX43-PARP1-NAD⁺ pathway's role in vascular aging and identify a potential therapeutic strategy to combat aging-associated BBB leakage with neuroprotective implications.