| Objective To screen for metabolites that differ between clinical cases and animal models by tandem mass spectrometry,and to explore the possible mechanisms by which whole-body vibration exercise prevents diabetic cardiomyopathy in mice by combining animal and cellular experiments,and to provide a theoretical basis for the clinical application of whole-body vibration exercise.Methods 1 、Differential metabolites were obtained by Partial Least SquaresDiscriminant Analysis(PLS-DA)of tandem mass spectrometry data from 30 nondiabetic patients and 30 diabetic patients.90 8-week-old C57BL/6J mice,30 randomly selected as blank controls and 60 fed with high-fat chow.60 mice were fed a high-fat diet with intraperitoneal injection of STZ to construct a type 2 diabetic mouse model,and their body weight and blood glucose were monitored regularly.The type 2 diabetic mice were then randomly divided into diabetic group and diabetic vibration group,30 mice in each group.The diabetic vibration group was given whole body vibration at a frequency of 15 Hz once a day for 30 minutes,5 days a week.After a total of 12 weeks of vibration,serum was taken from 10 randomly selected rats in each group for tandem mass spectrometry analysis.Differential metabolites were obtained by PLS-DA analysis.2、The effects of whole-body vibration on diabetic cardiomyopathy were analysed by body weight,blood glucose,oral glucose tolerance test(OGTT),oil red staining of myocardial sections,Western Blot for mTOR,S6K1,PGC1-α,CPT1B protein expression and immunofluorescence for PGC1-α and CPT1 B fluorescence intensity.3、The rat cardiomyocyte H9C2 was divided into normal,3m M arginine and 6m M arginine groups,and mTOR,S6K1,PGC1-α and CPT1 B protein expressions were detected by Western Blot to verify the effects of arginine in vitro.4、All count data were expressed as absolute numbers and relative ratios,and measures conforming to normal distribution were expressed as(?) ± s.The chi-square test,t-test and ANOVA were performed using SPSS 20.0 statistical software,and all metabolic mass spectrometry data were standardized and statistically analyzed using Metabo Analyst 5.0.*P < 0.05,**P < 0.01,***P < 0.001,****P < 0.0001.Results 1、Combined with the clinically collected data,it was found that C10,arginine,C5,C5:1 and ornithine could better differentiate the groups according to the Variable Importance in the Projection score(VIP score).Venn diagrams found a consistent trend in arginine in human samples versus mouse samples.Clinical data showed significantly lower arginine levels in diabetics than in the general population.Arginine levels were significantly lower in the diabetic group than in the control and diabetic vibration groups.2、The diabetic group showed increased body weight and blood glucose,increased area under the OGTT curve,inhibition of the mTOR pathway in myocardial tissue,decreased expression of PGC1-α and S6K1 and increased expression of CPT1 B compared to the control group.In the diabetic vibration group,compared to the diabetic group,body weight and blood glucose decreased,the area under the OGTT curve decreased,mTOR pathway activation in cardiac tissue,PGC1-α and S6K1 expression increased,and CPT1 B expression decreased.3 、 In cellular experiments,the higher the arginine content in the cardiomyocyte medium,the higher the level of mTOR phosphorylation,PGC1-α,S6K1 protein expression and the lower the CPT1 B protein expression.Conclusions 1、Serum arginine levels are low in diabetic patients and mice,whole body vibration exercise can increase arginine levels in diabetic mice.2、Whole Body Vibration exercise can improve glycemic control in diabetes,promote mitochondrial synthesis,promote protein synthesis,reduce lipid metabolism and decrease lipid accumulation.3、Arginine can stimulate the expression of downstream related proteins through activation of the mTOR pathway,promote cardiac mitochondrial biosynthesis,protein synthesis,reduce lipid metabolism and decrease lipid accumulation.. |
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