LncRNA-Fendrr protects against the ubiquitination and degradation of NLRC4 protein through HERC2 to regulate the pyroptosis of microglia

Autor: Pin Wu, Shengmei Zhu, Liqing Wang, Xiong-Xin Zhang, Heng-Jun Zhou, Yueying Zheng
Rok vydání: 2021
Předmět:
Male
0301 basic medicine
Cerebral ischemia–reperfusion
Biochemistry
0302 clinical medicine
Ubiquitin
Guanine Nucleotide Exchange Factors
Genetics (clinical)
Gene knockdown
Microglia
biology
medicine.diagnostic_test
Chemistry
Diabetes
Pyroptosis
Brain
Infarction
Middle Cerebral Artery
Ubiquitin ligase
Cell biology
medicine.anatomical_structure
Reperfusion Injury
Molecular Medicine
RNA
Long Noncoding
Research Article
RM1-950
QD415-436
Cell Line
Proinflammatory cytokine
LncRNA-Fendrr
03 medical and health sciences
Western blot
Diabetes Mellitus
Genetics
medicine
Animals
Molecular Biology
HERC2
Inflammation
NLRC4
Calcium-Binding Proteins
Ubiquitination
Mice
Inbred C57BL
030104 developmental biology
Cell culture
biology.protein
Therapeutics. Pharmacology
Apoptosis Regulatory Proteins
030217 neurology & neurosurgery
Zdroj: Molecular Medicine, Vol 27, Iss 1, Pp 1-10 (2021)
Molecular Medicine
ISSN: 1528-3658
1076-1551
DOI: 10.1186/s10020-021-00299-y
Popis: Objectives Targeted inhibition of inflammatory response can reduce diabetic cerebral ischemia–reperfusion (I/R) injure. Pyroptosis is characterized by caspase-1 dependence and the release of a large number of pro-inflammatory factors. LncRNA-Fendrr is associated with a variety of diseases, but Fendrr has not been studied in diabetic cerebral I/R. NLR-family CARD-containing protein 4 (NLRC4) regulate the pyroptosis of microglia cells. This study was designed to investigate whether Fendrr is involved in the effects of diabetic cerebral I/R injury. Methods The diabetic brain I/R model in mice was constructed. Mouse microglia cell line BV-2 cells were exposed to high glucose followed by hypoxia/reoxygenation (H/R). Fendrr and some pyroptosis-associated proteins were detected by qRT-PCR, western blot or ELISA. HE staining was used to detect pathological changes. Microglia pyroptosis was detected by TUNEL staining. RNA pull-down and RNA Immunoprecipitation were used to detect binding of Fendrr to HERC2 (E3 ubiquitin ligase), and CO-IP detected binding of HERC2 to NLRC4. The ubiquitination of NLRC4 was detected by ubiquitination experiments. Results Fendrr was significantly increased in the diabetic cerebral I/R model, and NLRC4 inflammatory complex and pyroptosis mediated inflammatory factors were increased. NLRC4 and inflammatory cytokines associated with pyroptosis were decreased in the high glucose-treated hypoxia/reoxygenation (H/R)-induced microglia after Fendrr knockdown. Fendrr bound to HERC2 protein, and HERC2 bound to NLRC4. Meanwhile, Fendrr could inhibit the ubiquitination of NLRC4, HERC2 promoted the ubiquitination of NLRC4 protein. Moreover, the effect of Fendrr overexpression in the diabetic cerebral I/R model of microglia can be reversed by HERC2 overexpression. Conclusion Fendrr can protect against the ubiquitination and degradation of NLRC4 protein through E3 ubiquitin ligase HERC2, thereby accelerating the pyroptosis of microglia.
Databáze: OpenAIRE
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