ULK1 Mediated Autophagy-Promoting Effects of Rutin-Loaded Chitosan Nanoparticles - NF-κB Signaling and EMT Inhibition Study

Sep 21, 2026

Uploaded by Eliana

Rutin-loaded chitosan nanoparticles (rCNPs) are evaluated for therapeutic potential against human hepatoma Hep3B cells, focusing on mechanisms that extend beyond epithelial–mesenchymal transition (EMT) inhibition. rCNPs are synthesized via ionic gelation, characterized by UV–Vis, FTIR, DLS, and TEM, and assessed for drug loading, pH-responsive release, cytotoxicity, and stability. Mechanistic studies indicate ULK1-mediated autophagy and NF-κB signaling activation, alongside increased ROS and mitochondrial membrane potential changes, with reduced toxicity toward L929 fibroblasts and strong hemocompatibility supporting intravenous cancer therapy feasibility.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages29

Reading Time1 h 13 min

AccessFree to download

Background

Methods

Results

Drug loading and release

Cytotoxicity and mechanistic findings

Biocompatibility and hemocompatibility

Conclusion

International Journal of Nanomedicine Dovepress open access to scientific and medical research Open Access Full Text Article ORIGINAL RESEARCH ULK1 Mediated Autophagy-Promoting Effects of Rutin-Loaded Chitosan Nanoparticles Contribute to the Activation of NF-κB Signaling Besides Inhibiting EMT in Hep3B Hepatoma Cells Peng Wu 1,*, Xiaoyong Wang 2,*, Min Yin 1,*, Wenjie Zhu3,*, Zheng Chen 1, Yang Zhang 1, Ziyu Jiang4, Longqing Shi 5, Qiang Zhu 1 1Children’s Hospital of Nanjing Medical University, Nanjing, People’s Republic of China; 2The People’s Hospital of Rugao, Nantong, People’s Republic of China; 3Kangda College of Nanjing Medical University, Nanjing, People’s Republic of China; 4Department of Oncology, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210028, People’s Republic of China; 5Department of Hepatobiliary and Pancreatic Surgery, Third Affiliated Hospital of Soochow University, Jiangsu, People’s Republic of China *These authors contributed equally to this work Correspondence: Longqing Shi, Department of Hepatobiliary and Pancreatic Surgery, The Third Affiliated Hospital of Soochow University, Jiangsu, People’s Republic of China, Email [20204135017@stu.suda.edu.cn](20204135017@stu.suda.edu.cn); Qiang Zhu, Children’s Hospital of Nanjing Medical University, Nanjing, People’s Republic of China, Email [zhu20081023@yeah.net](zhu20081023@yeah.net) Background: Liver cancer remains to be one of the leading causes of cancer worldwide. The treatment options face several challenges and nanomaterials have proven to improve the bioavailability of several drug candidates and their applications in nanomedicine. Specifically, chitosan nanoparticles (CNPs) are extremely biodegradable, pose enhanced biocompatibility and are considered safe for use in medicine. Methods: CNPs were synthesized by ionic gelation, loaded with rutin (rCNPs) and characterized by ultraviolet–visible spectroscopy (UV-Vis), Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and transmission electron microscopy (TEM) . The rCNPs were tested for their cytotoxic effects on human hepatoma Hep3B cells, and experiments were conducted to determine the mechanism of such effects. Further, the biocompatibility of the rCNPs was tested on L929 fibroblasts, and their hemocompatibility was determined. Results: Initially, UV–vis and FTIR analyses indicated the possible loading of rutin on rCNPs. Further, the rutin load was quantitatively measured using Ultra-Performance Liquid Chromatography (UPLC) and the concentration was 88 µg/mL for 0.22 micron filtered rCNPs. The drug loading capacity (LC%) of the rCNPs was observed to be 13.29 ± 0.68%, and encapsulation efficiency (EE%) was 19.55 ± 1.01% . The drug release was pH-responsive as 88.58% of the drug was released after 24 hrs at the lysosomal pH 5.5, whereas 91.44% of the drug was released at physiological pH 7.4 after 102 hrs. The cytotoxic effects were prominent in 0.22 micron filtered samples of 5 mg/mL rutin precursor. The particle size for the rCNPs at this concentration was 144.1 nm and the polydispersity index (PDI) was 0.244, which is deemed to be ideal for tumor targeting. A zeta potential (ζ-potential) value of 16.4 mV indicated rCNPs with good stability. The IC50 value for the cytotoxic effects of rCNPs on human hepatoma Hep3B cells was 9.7 ± 0.19 μg/mL of rutin load. In addition, the increased production of reactive oxygen species (ROS) and changes in mitochondrial membrane potential (MMP) were observed. Gene expression studies indicated that the mechanism for cytotoxic effects of rCNPs on Hep3B cells was due to the activation of Unc-51-like autophagy-activating kinase (ULK1) mediated autophagy and nuclear factor kappa B (NF-κB) signaling besides inhibiting the epithelial–mesenchymal Transition (EMT) . In addition, the rCNPs were less toxic on NCTC clone 929 (L929) fibroblasts in compariso

FAQs

How are rutin-loaded chitosan nanoparticles (rCNPs) prepared and characterized?

rCNPs are synthesized by ionic gelation and loaded with rutin. Characterization is performed using UV–Vis, FTIR, dynamic light scattering (DLS), and transmission electron microscopy (TEM).

What mechanism explains rCNPs’ effects on Hep3B hepatoma cells?

Gene expression results indicate that cytotoxic effects are due to ULK1-mediated autophagy and activation of NF-κB signaling, in addition to inhibiting EMT.

How do rCNPs perform in safety-related tests compared with normal cells?

rCNPs show less toxicity to L929 fibroblasts than to Hep3B cells and exhibit excellent hemocompatibility, with hemolysis below 2%, indicating suitability for intravenous administration.

Sep 21, 2026

Uploaded by Eliana