HECTD3 promotes NLRP3 inflammasome and pyroptosis to exacerbate diabetes-related cognitive impairment by stabilising MALT1 to regulate JNK pathway

Abstract

Background

HECTD3 (HECT domain E3 ubiquitin protein ligase 3) exerts biological activities in neuroinflammation of distinct diseases, such as autoimmune encephalomyelitis and donations after heart death. However, the effect of HECTD3 on diabetes-associated cognitive decline (DACD) remains unclear.

Methods

Wild-type or HECTD3-knockout rats were administered with streptozotocin to establish diabetic model. Pathological changes in the hippocampus were assessed by NISSL and haematoxylin and eosin staining. Morris water maze test was used to assess cognitive function. Neuronal survival and inflammation were investigated by immunofluorescence staining and ELISA assay. NLRP3 inflammasome and pyroptosis were assessed by western blot, immunofluorescence and flow cytometry assays.

Results

HECTD3 was up-regulated in hippocampus of streptozotocin-induced diabetic rats and high glucose-induced PC12 cells. Knockout of HECTD3 increased the number of neurons and improved the learning and memory function. Moreover, knockout of HECTD3 promoted in vivo neuronal survival, and reduced levels of IL-1β, TNF-α, and IL-6 in the hippocampus. Silencing of HECTD3 increased cell viability, and reduced IL-1β, TNF-α, and IL-6 in high glucose-induced PC12 cells. Fluorescence intensities of NLRP3, GSDMD-N and caspase-1 were reduced in HECTD3-knockout diabetic rats, and knockdown of HECTD3 down-regulated protein expression of NLRP3, GSDMD-N, caspase-1, IL-1β, and IL-18 in high glucose-induced PC12 cells to suppress the pyroptosis. HECTD3 promoted the stability of mucosa-associated lymphoid tissue 1 (MALT1) through up-regulation of c-JUN and phospho (p)-JNK in high glucose-induced PC12 cells. Over-expression of MALT1 attenuated neuroprotective effects of HECTD3 silencing on high glucose-induced PC12 cells.

Conclusion

HECTD3 silencing exerted neuroprotective effect against DACD through MALT1-mediated JNK signalling.

Highlights

• HECTD3 was up-regulated in hippocampus of streptozotocin-induced diabetic rats and high glucose-induced PC12.

• Knockout of HECTD3 promoted in vivo neuronal survival, reduced inflammation and pyroptosis, and improved the learning and memory function in diabetic rats.

• Knockout of HECTD3 suppressed the activation of NLRP3 inflammasome in diabetic rats.

• Silencing of HECTD3 exerted neuroprotective effects through MALT1-mediated JNK signalling.

Keywords:

• HECTD3
• diabetes-associated cognitive decline
• diabetes mellitus
• hyperglycaemia
• JNK pathway

Author contributions

Zhongfan Ruan designed the study, supervised the data collection, Yan Li analysed the data, interpreted the data, Yanfang Chen prepare the manuscript for publication and reviewed the draft of the manuscript. All authors have read and approved the manuscript.

Ethical approval

Ethical approval was obtained from the Ethics Committee of the First Affiliated Hospital, Hengyang Medical School, University of South China.

Disclosure statement

The authors state that there are no conflicts of interest to disclose.

Data availability statement

All data generated or analysed during this study are included in this published article.

Additional information

Funding

This work was supported by the Guiding project of Hengyang science and Technology Bureau [Grant No. 2020jh042748], Scientific Research Fund Project of Hunan Provincial Health Commission [Grant No. 20200151], Scientific research project of Hunan Provincial Department of Education [Grant No. 20C1634] and Scientific Research Fund Project of Hunan Provincial Health Commission [Grant No. 202203074376].