Abstract: Background　Antibiotic resistance is one of the main reasons for the failure to eradicate Helicobacter pylori (H. pylori) infection. Advances in the research of nanomaterials in the field of antimicrobials provide new ideas for the non-antibiotic treatment of H. pylori. Objective　This study examines how the magnetic hyperthermia generated by zinc ferrite nanoparticles (Zn0.3Fe2.7O4@SiO2) under an alternating magnetic field influences H. pylori growth and virulence. The underlying mechanism of this eradication was preliminarily explored through transcriptome analysis.Methods　H. pylori in logarithmic growth phase was spread onto Karmali 5% defibrinated sheep blood agar medium containing Zn0.3Fe2.7O4@SiO2 (50 μg/mL) and cultured in microaerophilic environment at 37℃ for 24 h. The medium was then exposed to an alternating magnetic field, heated to 41℃ for 15 min, and returned to microaerophilic environment, cultured at 37℃ for 72 h. The control group was treated by spreading an equal amount of H. pylori onto the surface of medium without Zn0.3Fe2.7O4@SiO2, and then cultured under microaerobic conditions at 37℃ for 72 hours without magnetic hyperthermia treatment. Following treatment, H. pylori was collected to examine the effects of magnetic hyperthermia on its growth, adhesion capacity, vacuolating cytotoxin activity, and urease activity, as well as to observe ultrastructural changes within H. pylori. Transcriptome sequencing, along with GO and KEGG pathway analyses, was conducted on the H. pylori. These transcriptomic results were further validated through qRT-PCR.Results　The growth level of H. pylori after magnetic hyperthermia decreased by 50.96% compared with the control group (P＜0.001), the adhesion ability decreased by 23.7% compared with the control group (P = 0.006), the vacuolating toxin activity decreased by 46.72% compared with the control group (P ＜0.001), and the urease activity decreased by 31.30% compared with the control group (P＜0.001). Transmission electron microscope revealed swelling, lysis, separation of the cell wall from the cytoplasm, uneven cytoplasmic distribution, reduced ribosome numbers, and irreversible ultrastructural damage. Transcriptomic analysis identified 326 DEGs, 119 DEGs upregulated, 207 DEGs downregulated. The expressions of urease-related genes such as ureE, ureD, ureG and ureA in DEGs were significantly downregulated. Enrichment analysis of downregulated genes identified 82 GO terms and 5 KEGG pathways, primarily linked to ribosomes and energy metabolism. qRT-PCR validation was consistent with the sequencing data. Conclusion　Magnetic hyperthermia inhibits H. pylori growth, attenuates its virulence, and alters its ultrastructure. Transcriptome analysis suggests that the underlying mechanism may be associated with energy metabolism, protein metabolism, and ribosomal structure and function.