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DDAH-1, via regulation of ADMA levels, protects against ischemia-induced blood-brain barrier leakage

Laboratory Investigation (2021)Cite this article

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Abstract

Dimethylarginine dimethylamino hydrolase-1 (DDAH-1) is an important regulator of nitric oxide (NO) metabolism that has been implicated in the pathogenesis of cardiovascular diseases. Nevertheless, its role in cerebral ischemia still needs to be elucidated. Herein, we examined the expression of DDAH-1 in the brain of rat by double-label immunofluorescence staining. DDAH-1 knock-out (DDAH-1−/−) and wild-type rats underwent middle cerebral artery occlusion/reperfusion (MCAO/R). After 24 h, neurological scores, TTC staining and TUNEL assay were used to evaluate neurological damages. 3 and 7-days infarct outcomes were also shown. Blood-brain-barrier (BBB) permeability was examined via Evans blue extravasation and tight junction (TJ) proteins expression and mRNA levels by western blot and RT-qPCR. The levels of plasma asymmetric dimethylarginine (ADMA), NO and ADMA in brain tissue were also assessed. In addition, supplementation of L-arginine to DDAH-1−/− rats was used to explore its role in regulating NO. DDAH-1 was abundantly distributed in cerebral cortex and basal nuclei, and mainly expressed in neurons and endothelial cells. DDAH-1−/− rats showed aggravated neurological damage and BBB disruption, including decrease of TJ proteins expression but indistinguishable mRNA levels after MCAO/R. DDAH-1 depletion and neurological damages were accompanied with increased ADMA levels and decreased NO concentrations. The supplementation with L-arginine partly restored the neurological damages and BBB disruption. To sum up, DDAH-1 revealed to have a protective role in ischemia stroke (IS) and IS-induced leakage of BBB via decreasing ADMA level and possibly via preventing TJ proteins degradation.

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Fig. 1: Main distribution of DDAH-1 in basal nuclei, not brainstem.
Fig. 2: Main distribution of DDAH-1 in neurons and endothelial cells.
Fig. 3: DDAH-1−/− rats showed aggravated ischemic damage after MCAO/R.
Fig. 4: DDAH-1−/− rats showed severer apoptosis in brain after MCAO/R.
Fig. 5: DDAH-1−/− rats showed increased EBD extravasation after MCAO/R with lower NO and higher ADMA levels.
Fig. 6: Supplement of L-arginine to DDAH-1−/− rats alleviated partial neurological damages.
Fig. 7: Supplement of L-arginine to DDAH-1−/− rats relieved long-term neurological damages.
Fig. 8: DDAH-1−/− rats showed decreased expressions of tight junction proteins.

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Acknowledgements

DDAH-1 knockout SD rats were provided by Professor Da-Chun Xu (Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine).

Funding

The present study was supported by the Science and Technology Commission of Shanghai Municipality (grant numbers 18140901900 and 20ZR1443500), National Natural Science Foundation of China (Grant no. 8207052336), and Shanghai Municipal Key Clinical Specialty (Grant no. shslczdzk06102).

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  1. Department of Neurology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, No.301 Middle Yanchang Road, Shanghai, 200072, PR China

    Yichen Zhao, Xiaoye Ma, Yuchen Zhou, Junchao Xie, Xueyuan Liu & Yanxin Zhao

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  1. Yichen Zhao
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Zhao, Y., Ma, X., Zhou, Y. et al. DDAH-1, via regulation of ADMA levels, protects against ischemia-induced blood-brain barrier leakage. Lab Invest (2021). https://doi.org/10.1038/s41374-021-00541-5

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