@Article{biocell.2024.055029,
AUTHOR = {TIANYUN HAN, ZHONG LI, LUONING ZHANG, LINSHEN XIE},
TITLE = {Ganoderic acid A ameliorates renal fibrosis by suppressing the expression of <i>NPC1L1</i>},
JOURNAL = {BIOCELL},
VOLUME = {48},
YEAR = {2024},
NUMBER = {11},
PAGES = {1625--1638},
URL = {http://www.techscience.com/biocell/v48n11/58592},
ISSN = {1667-5746},
ABSTRACT = { <b>Objective:</b> The study aimed to explore the protective mechanism of Ganoderic acid A (GAA) in renal fibrosis and to verify that GAA can ameliorate renal fibrosis by regulating the <i>Niemann-pick C1-like 1 (NPC1L1)</i> gene. <b>Methods:</b> Transforming growth factor beta1 (TGF-β1) was used to treat Human Kidney-2 (HK-2) cells to establish a renal fibrosis model. The differentially expressed genes in the control (CTRL) group, TGF-β1 group, and TGF-β1 + GAA group were screened via transcriptome sequencing technology and verified by qPCR and Western blot experiments. The <i>NPC1L1</i> gene overexpression plasmid was constructed. The expression levels of N-cad, E-cad, and Slug-related proteins in CTRL, TGF-β1, TGF-β1+GAA (25 μg/mL), and TGF-β1+GAA (25 μg/mL) + NPC1L1 Overexpression (OE) groups were detected by qPCR and Western blot analysis. Western blot analysis was used to identify the extracellular matrix-associated proteins Tenascin-C, α-SMA, and fibrosis-related protein Collagen I. Fibrosis marker protein Fibronectin was detected and quantified by immunofluorescence. <b>Results:</b> Transcriptomic sequencing revealed that TGF-β1 stimulation led to 267 differentially regulated genes, with 118 up-regulated and 149 down-regulated, while further modulation of 213 genes, comprising 112 up-regulated and 101 down-regulated genes, was observed in the GAA intervention group. The target gene in these processes was found to be <i>NPC1L1</i> by investigations using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). qPCR and Western blot results confirmed that TGF-β1 increased <i>NPC1L1</i> expression, which was attenuated by GAA. Additionally, TGF-β1 upregulated N-cad and Slug. However, GAA reversed this effect and <i>NPC1L1</i> overexpression partially rescued the GAA effect. TGF-β1 also decreased E-cad expression, reversed by GAA, and <i>NPC1L1</i> overexpression antagonized this reversal. Furthermore, TGF-β1 promoted Collagen I, α-SMA, and Tenascin-C expression, and GAA reduced these levels, effects that were reversed by <i>NPC1L1</i> overexpression. Immunofluorescence results showed that TGF-β1 increased fibronectin expression, which was decreased by GAA, and increased by <i>NPC1L1</i> overexpression. <b>Conclusion:</b> GAA ameliorates renal fibrosis by antagonizing <i>NPC1L1</i> gene expression inhibiting epithelial-mesenchymal transition and reducing extracellular matrix formation.},
DOI = {10.32604/biocell.2024.055029}
}