The Mechanism Of ZFYVE28 Involved In The Occurrence And Development Of Obesity And The Mechanism Of THA In The Treatment Of Arteriovenous Malformation

Background:Obesity may greatly increase the risk of type 2 diabetes,cancer and various cardiovascular diseases.Insulin signaling pathway can maintain energy metabolism balance in the body,and its disorder is an important factor leading to metabolic syndrome in obese patients.Currently,the mechanism of occurrence and development of obesity is still fully clear,and more in-depth researches are needed.Method:In this study,quantitative fluorescence PCR(QF-PCR)was performed on the fat and liver tissues of ob/ob obese mice with Leptin deficiency.The cause of abnormal expression of ZFYVE28 caused by obesity was studied through treating the simulated in vitro cell model of hyperinsulinemia in obese patients with high concentration of insulin.Afterwards,the mechanism of insulin regulating ZFYVE28 expression was studied based on HepG2 cells,and the function of ZFYVE28 was investigated.After ZFYVE28 was determined to be indirectly regulated by insulin signaling and can promote the degradation of activated insulin receptors,the correlation between ZFYVE28 and the expression of multiple receptor tyrosine kinase(RTKs)was analyzed by GEPIA.The correlation between ZFYVE28 and tumor prognosis and tumor proliferation were analyzed through various published cancer expression profiling databases.The expression of ZFYVE28 in various tumors was analyzed by Oncomine and TIMER.The effect of changes in ZFYVE28 expression level on the clinical prognosis of the tumor was then evaluated using the PrognoScan,Kaplan-Meier Plotter and GEPIA.Whether the expression level of ZFYVE28 could change the infiltration of immune cells inside the tumor was next investigated using the TIMER.Finally,the hypothesis that ZFYVE28 was involved in tumor proliferation was verified by established tumor-bearing mouse models.Results:ZFYVE28 mRNA level in the fat and liver of obese mice was increased significantly.The experimental results based on the cell model showed that the insulin signal could down-regulate ZFYVE28 expression.The effect of insulin signal on inhibiting ZFYVE28 expression was limited after insulin resistance was induced to block the activation of the insulin signaling pathway.Inhibitors added with RAS signal were found to be able to up-regulate the expression of ZFYVE28 in subsequent experiments,but the same effect could not be produced after using similar methods to cut off AKT signal.NOTCH-mediated transcription initiation of ZFYVE28 was inhibited by insulin signaling through promoting the degradation of NOTCH intracellular domain(NICD).In addition,ZFYVE28 could promote the degradation of insulin receptor after activation and participate in the formation of insulin resistance.Through the correlation analysis of gene expression in tumor tissues,the expression level of ZFYVE28 was found to be significantly related to the expression of various RTKs involved in many biological processes such as regulating cell proliferation.After analyzing Oncomine and TIMER,the expression level of ZF YVE2 8 in tumor tissue was found to be significantly lower than that of paracancerous tissue.After analyzing the RNA sequencing data of Pan-Cancer cohort in Kaplan-Meier Plotter,the high expression of ZFYVE28 showed a correlation with better overall survival(OS)(HR=0.7,P=0.024).Combined with the analysis of TCGA and PrognoScan,ZFYVE28 was found to be significantly related to the clinical prognosis of various tumors.Finally,ZFYVE28 was proven from animal experiments to inhibit tumor proliferation in vivo.Conclusion:ZFYVE28 is confirmed in the study to be regulated by insulin signal feedback and can promote the degradation of activated insulin receptors,indicating that ZFYVE28 is involved in the formation of insulin resistance.Meanwhile the function of ZFYVE28 to promote receptor degradation may affect a variety of RTKs,but the mechanism needs to be experimentally verified in the future.ZFYVE28 is closely related to the clinical prognosis of cancer patients and can affect tumor proliferation in vivo.In short,the mechanism of ZFYVE28's involvement in the occurrence and development of obesity is deeply studied in this research.The results indicate that ZFYVE28 is involved in the formation of insulin resistance and is a key factor leading to an increased tumor incidence in obese patients.Cerebral arteriovenous malformation is one of the important risk factors for intracranial hemorrhage in young people.There are currently considerable risks in the treatment of cerebral arteriovenous malformations,and safe treatments in preventing rupture and bleeding are still unavailable.Sporadic cerebral arteriovenous malformations are mainly caused by KRAS/BRAF gain-of-function mutations,which may downregulate the expression of PDGFB in endothelial cells,leading to incomplete SMC recruitment.THA has been demonstrated in this study to inhibit the migration activity of primary endothelial cells isolated from the focal site of patients with sporadic cerebral arteriovenous malformations and promote their SMC recruitment.After THA treatment,the PDGFB expression in endothelial cells is significantly increased,and the ability of SMC recruitment is significantly improved.The effect of THA on endothelial cells has been also found to be reflected in the down-regulation of MYC expression and the increase of FN expression in the in-depth mechanism study.These changes can increase the patient's vascular stability in different ways.These results provide a theoretical basis for the clinical application of THA in the treatment of sporadic cerebral arteriovenous malformations,and are of great significance for promoting the development of drugs that target arteriovenous malformations.