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Long Non-coding RNA H19 Promotes NLRP3-Mediated Pyroptosis After Subarachnoid Hemorrhage in Rats

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Abstract 

NLRP3 inflammasomes have been reported to be an essential mediator in the inflammatory response during early brain injury (EBI) following subarachnoid hemorrhage (SAH). Recent studies have indicated that NLRP3 inflammasome-mediated pyroptosis and long non-coding RNA (lncRNA) H19 can participate in the inflammatory response. However, the roles and functions of lncRNA H19 in NLRP3 inflammasome-mediated pyroptosis during EBI after SAH are unknown and need to be further elucidated. NLRP3 inflammasome proteins were significantly elevated in CSF of human with SAH induced EBI and presented a positive correlation with severity. In ipsilateral hemisphere cortex of rats, these NLRP3 inflammasome proteins were also increased and accompanied with upregulation of H19, and both of NLRP3 and H19 were peaked at 24 h after SAH. However, knockdown of H19 markedly decreased the expression of NLRP3 inflammasome proteins at 24 h after SAH in rats and also ameliorated EBI, showing improved neurobehavioral deficits, cerebral edema, and neuronal injury. Moreover, knocking down of H19 downregulated the expression of Gasdermin D (GSDMD) in microglia in SAH rats. Similarly, knockdown of H19 also alleviated OxyHb-induced pyroptosis and NLRP3-mediated inflammasomes activation in primary microglia. Lastly, H19 competitively sponged with rno-miR-138-5p and then upregulated NLRP3 expression in the post-SAH inflammatory response. lncRNA H19 promotes NLRP3-mediated pyroptosis by functioning as rno-miR-138-5p sponge in rats during EBI after SAH, which might provide a potential therapeutic target for post-SAH inflammation regulation.

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Abbreviations

NLRP3 :

Nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing 3

EBI :

Early brain injury

SAH :

Subarachnoid hemorrhage

lncRNAs :

Long non-coding RNAs

CSF :

Cerebrospinal fluid

IL :

Interleukin

ICV :

Intracerebroventricular

AAV :

Adeno-associated virus

OxyHb :

Oxyhemoglobin

GSDMD :

Gasdermin D

ceRNAs :

Competitive endogenous RNAs

CT :

Computed tomography

DSA :

Digital subtraction angiography

mFS :

Modified Fisher scale

LD :

Lumbar drainage

EVD :

External ventricular drainage

CNS :

Central nervous system

BMI :

Body mass index

KD :

Knock down

CON :

Control

PCD :

Programmed cell death

FL :

Full length

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Acknowledgements

We are grateful to our researchers, collaborators, and technical assistants for their helpful suggestions and experimental assistance.

Funding

This study was supported by grants from National Natural Science Foundation of China (81870916) of Jianmin Zhang and National Natural Science Foundation of China (81971107) of Sheng Chen.

Author information

Author notes

  1. Dr. Yibo Liu, Yujie Luo, and Anke Zhang contributed equally to this work.

Authors and Affiliations

  1. Department of Neurosurgery, School of Medicine, The Second Affiliated Hospital, Zhejiang University, Zhejiang Province, Hangzhou, People’s Republic of China

    Yibo Liu, Yujie Luo, Anke Zhang, Zefeng Wang, Xiaoyu Wang, Qian Yu, Zeyu Zhang, Zhoule Zhu, Kaikai Wang, Jianmin Zhang, Yuanjian Fang & Sheng Chen

  2. Key Laboratory of Precise Treatment and Clinical Translational Research of Neurological Diseases, Hangzhou, Zhejiang, China

    Yibo Liu, Yujie Luo, Anke Zhang, Zefeng Wang, Xiaoyu Wang, Qian Yu, Zeyu Zhang, Zhoule Zhu, Kaikai Wang, Jianmin Zhang, Yuanjian Fang & Sheng Chen

  3. Division of Neurosurgery, Department of Surgery, Prince of Wales Hospital, The Chinese University of Hong Kong, Ma Liu Shui, Hong Kong, China

    Yujie Luo

  4. Department of Medical Genetics, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China

    Luxi Chen

  5. Department of Neurosurgery, The Affiliated Huzhou Hospital, Zhejiang University School of Medicine (Huzhou Central Hospital), Huzhou, China

    Xiaohu Nie & Zhongzhou Su

  6. Department of Neurosurgery, Loma Linda University, Loma Linda, CA, USA

    John H. Zhang

  7. Department of Physiology and Pharmacology, Loma Linda University, Loma Linda, CA, USA

    John H. Zhang

  8. Department of Anesthesiology, Loma Linda University, Loma Linda, CA, USA

    John H. Zhang

Authors
  1. Yibo Liu
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Contributions

Sheng Chen, Zhongzhou Su, and Yuanjian Fang contributed to the design of study. Yuanjian Fang, Xiaoyu Wang, and Zhoule Zhu performed the clinical sample collection and statistical analysis. Yibo Liu, Yujie Luo, Anke Zhang, Zefeng Wang, and Zeyu Zhang conducted the in vivo experiments. Yibo Liu, Qian Yu, and Kaikai Wang conducted the in vitro experiments. Yibo Liu, Luxi Chen, and Xiaohu Nie wrote the manuscript. John H. Zhang and Jianmin Zhang verified the underlying data. All authors read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Yuanjian Fang, Zhongzhou Su or Sheng Chen.

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Liu, Y., Luo, Y., Zhang, A. et al. Long Non-coding RNA H19 Promotes NLRP3-Mediated Pyroptosis After Subarachnoid Hemorrhage in Rats. Transl. Stroke Res. 14, 987–1001 (2023). https://doi.org/10.1007/s12975-022-01104-6

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