RVG-EVs-mediated Delivery of siRNA Targeting circHIPK3 Attenuates Microglial M1 Polarization by Enhancing Mitophagic Flux
10.13865/j.cnki.cjbmb.2025.09.1167
- VernacularTitle:RVG-EVs递送靶向circHIPK3的siRNA通过增强线粒体自噬流抑制小胶质细胞M1型极化
- Author:
Yu YANG
1
;
Na DONG
;
Chi ZHANG
;
Zhen-Zhen HU
Author Information
1. 南京医科大学第一附属医院《江苏医药》编辑部,南京 210029
- Publication Type:Journal Article
- Keywords:
circular RNA HIPK3(circHIPK3);
microglia;
polarization;
mitophagy
- From:
Chinese Journal of Biochemistry and Molecular Biology
2025;41(11):1719-1728
- CountryChina
- Language:Chinese
-
Abstract:
Microglia activation-mediated neuroinflammatory responses serve as a critical pathological ba-sis for the development and progression of various brain diseases.The role of circular RNAs(circRNAs)in the regulation of neuroinflammation is increasingly being recognized.This study aimed to investigate the effect and molecular mechanisms of targeted inhibition of circular RNA Homeodomain Interacting Pro-tein Kinase 3(HIPK3)(circHIPK3)on lipopolysaccharide(LPS)-induced microglial polarization in BV2 cells.The results showed that LPS stimulation significantly induced polarization of BV2 cells towards the pro-inflammatory M1 phenotype and upregulated circHIPK3 expression(P＜0.01).Engineered extra-cellular vesicles(EVs)with rabies viral glycoprotein(RVG)loaded with circHIPK3 siRNA(RVG-EVs-sicHIPK3)were successfully constructed.Transmission electron microscopy(TEM)revealed their typi-cal EV morphology.nanoparticle tracking analysis(NTA)indicated a peak particle size of 70 nmn.And Western blotting analysis confirmed the expression of characteristic membrane marker proteins.Treatment with RVG-EVs-sicHIPK3 significantly suppressed the LPS-induced elevation of inflammatory cytokines(TNF-α,IL-6,IL-1β)in the supernatant and reduced the expression of M1 phenotypic marker proteins(CD16 and CD86)(P＜0.01).Concurrently,RVG-EVs-sicHIPK3 increased the number of mitophago-somes within cells,upregulated the ratio of the autophagy-related proteins LC3-Ⅱ/LC3-I(P＜0.01),and downregulated the expression of the autophagy-related protein p62 and mitochondrial-specific proteins(TOMM20 and TIMM23)(P＜0.01).The mitophagy inhibitor Mdivi-1 significantly reversed the RVG-EVs-sicHIPK3-mediated downregulation of inflammatory cytokine levels,M1 marker proteins,and mito-chondrial protein expression(P＜0.01).This study demonstrates that inhibiting circHIPK3 reduces LPS-induced microglial polarization towards the M1 phenotype.The protective mechanism is closely associated with enhanced mitophagic flux and the promotion of damaged mitochondrial clearance.
