Neuroprotection of NAD+ and NBP against ischemia/reperfusion brain injury is associated with restoration of sirtuin-regulated metabolic homeostasis

Xin-Xin Wang; Guang-Hui Mao; Qi-Qi Li; Jie Tang; Hua Zhang; Kang-Lin Wang; Lei Wang; Hong Ni; Rui Sheng; Zheng-Hong Qin

doi:10.3389/fphar.2023.1096533

Neuroprotection of NAD⁺ and NBP against ischemia/reperfusion brain injury is associated with restoration of sirtuin-regulated metabolic homeostasis

Front Pharmacol. 2023 Mar 28:14:1096533. doi: 10.3389/fphar.2023.1096533. eCollection 2023.

Authors

Xin-Xin Wang^{1

2}, Guang-Hui Mao¹, Qi-Qi Li¹, Jie Tang¹, Hua Zhang¹, Kang-Lin Wang³, Lei Wang³, Hong Ni², Rui Sheng¹, Zheng-Hong Qin¹

Affiliations

¹ Department of Pharmacology and Laboratory of Aging and Nervous Diseases, Jiangsu Key Laboratory of Neuropsychiatric Diseases, College of Pharmaceutical Sciences of Soochow University, Suzhou, China.
² Institute of Pediatric Research, Children's Hospital of Soochow University, Suzhou, China.
³ Hefei Knature Bio-pharm Co., Ltd., Hefei, China.

Abstract

Background: Ischemic stroke seriously threatens human health because of high rates of morbidity, mortality and disability. This study compared the effects of nicotinamide adenine dinucleotide (NAD⁺) and butylphthalide (NBP) on in vitro and in vivo ischemic stroke models. Methods: Transient middle cerebral artery occlusion/reperfusion (t-MCAO/R) model was established in mice, and the cultured primary cortical neurons were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R). Cerebral infarct volume, neurobehavioral indices, antioxidant activity, ATP level and lactic acid content were determined. The neuroprotective effects of NAD⁺ or NBP were compared using sirtuin inhibitor niacinamide (NAM). Results: Intraperitoneal injection of NBP within 4 h or intravenous injection of NAD⁺ within 1 h after t-MCAO/R significantly reduced the volume of infarcts, cerebral edema, and neurological deficits. Administration of NAD⁺ and NBP immediately after t-MCAO/R in mice showed similar neuroprotection against acute and long-term ischemic injury. Both NAD⁺ and NBP significantly inhibited the accumulation of MDA and H₂O₂ and reduced oxidative stress. NAD⁺ was superior to NBP in inhibiting lipid oxidation and DNA damage. Furthermore, although both NAD⁺ and NBP improved the morphology of mitochondrial damage induced by ischemia/reperfusion, NAD⁺ more effectively reversed the decrease of ATP and increase of lactic acid after ischemia/reperfusion compared with NBP. NAD⁺ but not NBP treatment significantly upregulated SIRT3 in the brain, but the sirtuin inhibitor NAM could abolish the protective effect of NAD⁺ and NBP by inhibiting SIRT1 or SIRT3. Conclusions: These results confirmed the protective effects of NAD⁺ and NBP on cerebral ischemic injury. NBP and NAD⁺ showed similar antioxidant effects, while NAD⁺ had better ability in restoring energy metabolism, possibly through upregulating the activity of SIRT1 and SIRT3. The protection provided by NBP against cerebral ischemia/reperfusion may be achieved through SIRT1.

Keywords: NAD+; NBP; SIRT1; SIRT3; acetylation; cerebral ischemia-reperfusion; mitochondria.

Grants and funding

This work was supported by the Natural Science Foundation of China (No. 81730092, 81973315 and 82173811), Natural Science Foundation of Jiangsu Higher Education (20KJA310008), Jiangsu Key Laboratory of Neuropsychiatric Diseases (BM2013003) and the Priority Academic Program Development of the Jiangsu Higher Education Institutes (PAPD).