Work

FTO mediates the diabetic kidney disease progression through regulating the m6A modification of NLRP3

Year: 2024

Type: article

Abstract: The objective of our research was to investigate the specific mechanism of FTO in diabetic kidney disease (DKD) progression. The DKD model was established with renal tubular epithelial HK-2 cells and mice in vitro and in vivo. The N6-methyladenosine (m 6 A) content in cells was detected using dot plot assay and the m 6 A levels of NLRP3 was detected with the MeRIP assay. The mRNA and protein levels were tested with real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and western blot. The IL-1β and IL-18 levels were assessed with enzyme-linked immunosorbent assay (ELISA). The cell viability was measured by cell counting kit (CCK)-8 assay and cell pyroptosis was determined with Annexin V and propidium iodide (PI) double staining followed by flow cytometry analysis. RNA-binding protein immunoprecipitation (RIP) and dual luciferase reporter assays were conducted to detect the interaction between FTO and NLRP3 . m 6 A levels were detected by Me-RIP assay. The renal injury was measured by observing the renal morphology and urine and blood levels of relevant indicators. The results indicated that high glucose treatment induced HK-2 cell pyroptosis. m 6 A levels were prominently elevated in high glucose treated HK-2 cells while FTO expression were significantly down-regulated. FTO over-expression promoted cell viability but inhibited pyroptosis of HK-2 cells under high glucose (HG) treatment. Moreover, FTO could inhibit NLRP3 expression. RIP and Me-RIP assays indicated that FTO could bind with NLRP3 and regulate its m 6 A modification level. Further luciferase assay confirmed that FTO binds with the 233–237 bp region of NLRP3 . NLRP3 neutralized the function of FTO in the HG stimulated HK-2 cells. In vivo, the H&E staining showed that FTO over-expression alleviated the kidney injury and suppressed the pyroptosis induced by DKD. We found that FTO could inhibit the DKD progression in vivo and in vitro by regulated the m 6 A modification of NLRP3 .

Source: BMC Nephrology

DOI: 10.1186/s12882-024-03741-5

Authors: Qiang Li, Shujuan Mu

Institutions: China Academy of Chinese Medical Sciences, Guang’anmen Hospital

Language: English

Topic: RNA modifications and cancer

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $2,890