Csi-Bantam Delivered by Clonorchis sinensis Extracellular Vesicles Promotes Liver Fibrosis via Targeting TRIM35

Freshwater fish is widely consumed, yet raw or undercooked freshwater fish can transmit foodborne zoonotic pathogens. Clonorchiasis, caused by Clonorchis sinensis, is a major foodborne parasitic disease associated with hepatobiliary injury and fibrosis. The biological functions and mechanisms underlying liver injury mediated by C. sinensis-derived extracellular vesicles (EVs) encapsulated microRNAs in fibrogenic remodeling remain poorly defined. Here, we isolated and characterized EVs from adult C. sinensis, confirmed their uptake by hepatic stellate cells (HSCs), and identified Csi-bantam as a CsEV-enriched microRNA. Csi-bantam enhances HSC activation and upregulates fibrogenic markers, including α-SMA, COL1A1, and COL3A1, both in vitro and in a mice infection model. Bioinformatic prediction and dual-luciferase assays validated TRIM35 as a target of Csi-bantam. TRIM35 silencing phenocopied Csi-bantam overexpression and increased PI3K/AKT phosphorylation, whereas Csi-bantam inhibition restored TRIM35 and attenuated fibrogenic markers. Collectively, CsEV-associated Csi-bantam promotes HSC activation via the TRIM35-PI3K/AKT axis, suggesting a potential target for antifibrotic intervention in clonorchiasis. These findings provide mechanistic insight into how infection acquired from raw or undercooked freshwater fish harboring infective C. sinensis metacercariae can lead to hepatobiliary injury and fibrogenic remodeling, supporting food safety risk assessment and future prevention of fish-borne parasitic hazards.