Lipopolysaccharide Promotes Dedifferentiation Of β Cells Induced By FTO/NLRP3 Pathway Activated By High Glucos

Background and ObjectiveType 2 diabetes mellitus(T2DM)is a metabolic disorder that affects a large and growing population.While there are multiple factors that contribute to T2DM,the loss of β cell mass due to β cell dedifferentiation is one of the key mechanisms underlying disease progression.However,current treatments cannot effectively alleviate β cell dysfunction and reverse dedifferentiation.Lipopolysaccharide(LPS)is a complex inflammatory mediator that is considered one of the important initiating factors of T2DM.NLRP3,as an inflammation signaling platform in the body,can respond to LPS signals to release IL-1β and IL-18,causing sustained inflammation and further participating in β cell dedifferentiation.In addition,the cascade mechanism between FTO and NLRP3 is gradually being revealed,and there is a proportional relationship between FTO expression and NLRP3 assembly.Meanwhile,with further research,FTO’s important pathogenic role in the T2DM disease process has been discovered.Although studies have identified FTO,NLRP3,and β cell dedifferentiation as important pathogenic factors in T2DM,there is still a lack of in-depth research on their interactions.Therefore,understanding the relationship between FTO and NLRP3 and pancreatic islet dedifferentiation has potential significance for the treatment and prevention of T2DM.This study aims to explore the expression of FTO/NLRP3 in T2DM models under LPS stimulation and further analyze whether FTO/NLRP3 has an inducing effect on β cell dedifferentiation.Through these studies,we hope to provide new opportunities for the clinical diagnosis and treatment of T2DM,and even hope to slow down or even reverse the progression of the disease.Methods1、Cell experimentation:INS-1 cell injury was induced by high glucose and LPS.Islet injury was evaluated by CCK8,KSIS and ELISA.Intracellular FTO,NLRP3,PDX1,OCT3/4 gene expression were detected by QPCR.Intracellular FTO,NLRP3,PDX1,OCT3/4 protein expression were detected by Western blot and cellular immunofluorescence assay.2、Animal experimentation:The rat model of T2DM was induced by a fat-rich diet combined with streptozotocin(STZ).The another group of T2DM rats was randomly chosen for the addition of LPS stimulation.Body weight,diet,drinking water,blood glucose,IPGTT and IPITT;HE staining were used to observe the pathological changes of the pancreas,and the IHC method was used to explore the expression of FTO,NLRP3,PDX1 and OCT3/4 in the pancreas.3、Bioinformatic analysis:the human pancreatic chip from GSE118139 was selected and RStudio was used for data qualification testing,analysis of gene expression differences,data enrichment and interaction analysis.ResultsElevated blood sugar and LPS can damage the functioning of INS-1 cells.The QPCR and Western Blot experiments showed that FTO,NLRP3,OCT3/4 were upward regulated and the expression PDX1 was inhibited.The rats in the experimental group had typical clinical manifestations of diabetes,blood glucose and insulin secretion reached the diagnostic criteria of diabetes,and the morphology and arrangement of islet β cells were damaged.QPCR,Western blot and other experiments demonstrated that it regulated FTO,NLRP3,OCT3/4 and inhibited PDX1 expression.Bioinformatics suggests that the expression of NLRP3 and OCT3/4 in human T2DM islands is controlled and has the potential for interaction.Enrichment analysis predicts a number of molecular processes and related signal pathways,providing valuable insights for future research.At the same time,through pathway analysis,it is inferred that there is a potential for FTO activation in human samples.ConclusionThis study found that FTO can activate NLRP3 inflammatory bodies and then stimulate the dedifferentiation of islet β cells.We also observed that under the influence of LPS,the expression of FTO increased and activated a large number of NLRP3 inflammatory bodies,at the same time,there is more obvious dedifferentiation of β cells.It is suggested that FTO plays an important role in the progression of T2DM disease and is expected to be a new target for inhibiting islet dedifferentiation in patients with T2DM in the future.