Honeysuckle-Derived Exosome-Like Nanovesicles Protect Against Acute Liver Failure by Modulating Gut Microbiota

Ping Li,1– 3 Yan Tang,1,3,4 Yixun Chen,1,3,4 Weijiao Fan,1,3 Jiayu Yao,1,3 Kexin Yu,1,3 Yiyi Shan,1,3 Jie Wang,5 Xiang Ming Ye,1 Hai Zou,6 Xiaozhou Mou1,3 1Center for Rehabilitation Medicine, Rehabilitation and Sports Medicine Research Institute of Zhejiang Province, Department of Rehabilitation Medicine, Zhejiang Provincial People’s Hospital (Affiliated People’s Hospital), Hangzhou Medical College, Hangzhou, People’s Republic of China; 2College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, People’s Republic of China; 3Clinical Research Institute, Zhejiang Provincial People’s Hospital (Affiliated People’s Hospital), Hangzhou Medical College, Hangzhou, People’s Republic of China; 4College of Life Sciences, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 5EVital Bio (Hangzhou) Co., Ltd, Hangzhou, People’s Republic of China; 6Department of Critical Care, Shanghai Cancer Center, Fudan University, Shanghai, People’s Republic of ChinaCorrespondence: Hai Zou, Email [email protected] Xiaozhou Mou, Email [email protected]: Acute liver failure (ALF) is a rare but life-threatening condition caused by drug toxicity, viral infections, or autoimmune disorders. Current treatments rely heavily on liver transplantation, which is costly and high-risk. As most therapies target single mechanisms, developing safe, cost-effective multi-target drugs is urgently needed.Methods: In this study, we isolated exosome-like nanovesicles from dried honeysuckle (HNVs) and evaluated their therapeutic potential in a lipopolysaccharide/D-galactosamine (LPS/GalN)-induced ALF mouse model. We first characterized HNVs using cryo-electron microscopy (Cryo-EM), transmission electron microscopy (TEM), and dynamic light scattering (DLS), and confirmed their stability in gastrointestinal simulation fluid in vivo. Subsequently, we validated the biological safety and in vivo distribution of HNVs in mice. Afterwards, we constructed an ALF model and tested the therapeutic efficacy of HNVs on this model. Through RNA sequencing, 16S rRNA analysis, and complementary techniques such as Western blot and quantitative real-time PCR, we elucidated the underlying mechanisms of HNVs in mitigating ALF.Results: Our results showed that HNVs significantly ameliorated the pathological symptoms associated with ALF mice. Specifically, HNVs induced a significant 1.89-fold decrease in serum ALT levels and a 1.95-fold decrease in AST levels. HNVs also ameliorated the hepatocellular necrosis and inflammatory cell infiltration caused by LPS/GalN. In addition, our findings suggest that the mechanism by which HNVs ameliorate ALF is: (1) they directly target the liver by traversing the compromised intestinal barrier, suppressing hepatic immune-inflammatory responses, and ameliorating ALF; (2) they restore intestinal barrier integrity by modulating the gut microbiota, thus reducing the translocation of gut-derived LPS to the liver and preventing further hepatic injury.Conclusion: In summary, we developed a novel natural nanomedicine with dual-targeting capabilities and demonstrated its efficacy against ALF, offering a promising therapeutic alternative for this severe condition.Keywords: acute liver failure, Honeysuckle-derived exosome-like nanovesicles, gut microbiota, immune-inflammatory responses