Tehran University of Medical Sciences Iranian Journal of Allergy, Asthma and Immunology 1735-1502 24 3 2025 05 05 361 374 You Wang Department of Nephrology, Wuhan Third Hospital, Tongren Hospital of Wuhan University, Wuhan, Hubei, China Pei Pi Department of Nephrology, Wuhan Third Hospital, Tongren Hospital of Wuhan University, Wuhan, Hubei, China Manli Hu Department of Nephrology, Wuhan Third Hospital, Tongren Hospital of Wuhan University, Wuhan, Hubei, China Dan Luo Department of Nephrology, Wuhan Third Hospital, Tongren Hospital of Wuhan University, Wuhan, Hubei, China 2024 11 19 2024 12 07 Latent transforming growth factor-β binding protein-2 (LTBP2) plays a significant role in tissue fibrosis. This research aimed to elucidate whether LTBP2 influences the progression of diabetic nephropathy (DN) through the phosphatidylinositol 3-kinases/protein kinase B (PI3K/Akt)/nuclear factor kappa-B (NF-κB) pathway. The HBZY-1 cells were exposed to high glucose to create diabetic nephropathy cell model. LTBP2 levels were examined by Western blot and immunofluorescence. After verifying the transfection efficiency of si-LTBP2, cell counting kit-8, 5-ethynyl-2-deoxyuridine staining, Western blot, flow cytometry and immunofluorescence were utilized to assess the proliferation, apoptosis and fibrosis of HBZY-1 cells, respectively. Collagen deposition was also detected by Sirius red staining, and inflammatory factors levels were determined by Elisa. PI3K/Akt/NF-κB pathway activators were applied to explore whether LTBP2 silencing could play a role in DN by modulating this pathway. After treatment with high glucose, the expression of LTBP2 was elevated in HBZY-1 cells. LTBP2 silencing hindered the aberrant proliferation of HBZY-1 cells, with no significant effect on apoptosis; meanwhile, it reduced fibrosis, decreased collagen content, and decreased inflammatory factors levels in HBZY-1 cells. Following treatment with high glucose, the PI3K, Akt, and p65 phosphorylation levels were increased, whereas silencing LTBP2 reduced them. Activators of the PI3K/Akt/NF-κB pathway weakened the inhibition of LTBP2 silencing on cell proliferation, fibrosis, and inflammation. In conclusion, silencing of LTBP2 weakened the proliferation, fibrosis, and inflammation of HBZY-1 cells treated with high glucose by hindering the PI3K/Akt/NF-κB pathway. This research offers a new reference for the targeted therapy of DN. https://ijaai.tums.ac.ir/index.php/ijaai/article/view/4247 https://ijaai.tums.ac.ir/index.php/ijaai/article/download/4247/2181