The Effect Of Blood Glucose Variability On Arterial Calcification And Supramolecular Peptides Pte-GFFpY Intervention

Background and Objective:Arterial calcification is a common vascular disease that occurs in many diseases,such as diabetes,chronic kidney disease and aging.It seriously threatens human health and causes heavy social and economic burden.However,there is still a lack of effective treatment for arterial calcification.Thus,early identification of potential risk factors and effective treatment strategies are important directions for the prevention and treatment of arterial calcification in the future.Epidemiological evidence show that fasting glucose variability(FGV)is closely related to the increased risk of cardiovascular events.However,the relationship between FGV and coronary artery calcification(CAC)remains unclear.Therefore,in the first part of our study,the relationship between FGV and CAC progression was analyzed by describing the risk factors of different FGV levels and the distribution of CAC in young and middle-aged people.In addition,recent studies have shown that alkaline phosphatase(ALP)in the lesion site of arterial calcification is significantly increased.Enzyme responsive self-assembled peptide is a new type of drug carrier.Due to its advantages of simple synthesis,high yield,good biocompatibility and biodegradability,its future application in clinical treatment has been widely concerned.Our previous study found that pterostilbene(Pte)has a good vascular protective effect.In order to explore the potential intervention strategy of arterial calcification,in the second and third part of this study,we prepared a small molecular polypeptide Pte-GFFpY loaded with Pte based on ALP response,and study the effect of Pte-GFFpY on arterial calcification and its potential mechanism.Methods and ResultsIn the first part of this study,2256 subjects from Coronary Artery Risk Development Study in Young Adults were included and followed up for 10 years.The 10-year CAC progress of each subject was assessed by coronary CT scaning.FGV was evaluated by fasting blood glucose coefficient of variation(FG-CV),standard deviation(FG-SD)and average real variability(FG-ARV).Multiple linear regression was used to analyze the association between FGV and CAC progression.After multivariable adjustment,1-SD increment in FG-CV was associated with worse progression of CAC as demonstrated as percent change in CAC with incident CAC 5.9%(95%CI 1.0,10.7)and any CAC progression 6.8%(95%CI 2.2,11.4)during 10 years.Similar results were also observed in FG-SD and FG-ARV.In the second and third part of this study,1)We have prepared a small polypeptide Pte-GFFpY.The results of mass spectrometry and transmission electron microscopy showed that Pte-GFFpY could self assemble to form nanofiber hydrogel under the action of ALP and stable release Pte.2)We established a mouse model of arterial calcification and evaluated the effect of Pte-GFFpY on arterial calcification by micro-CT and von kossa staining.The results showed that Pte-GFFpY could inhibit the progression of arterial calcification in mice.3)The calcification models of arterial rings and vascular smooth muscle cells(VSMC)were established in vitro.The degree of calcification was evaluated by alizarin red staining,von Kossa staining and calcium content detection.RT-qPCR,immunofluorescence and western blot were used to detect the gene and protein expression of VSMC contractile and osteogenic phenotype.The level of reactive oxidation products were detected by flow cytometry.These results showed that Pte-GFFpY could improve arterial rings calcification,inhibit the osteogenic differentiation and calcification of VSMC,and reduce the level of reactive oxidation products.Conclusion(1)Higher FGV during young adulthood was associated with greater CAC progression in middle age,which was independent of the average fasting blood glucose level and other traditional cardiovascular risk factors.(2)We have prepared a small polypeptide Pte-GFFpY which can self assemble in calcification site.(3)Pte-GFFpY can effectively inhibit the phenotype transdifferentiation and calcification of VSMC,and subsequent improve the progress of arterial calcification via downregulating the level of oxidative stress.