Serum Metabolomics Research Of MarvelD1 Knockout Mice Under Oxidative Stress

MarvelD1(MAL and related proteins for vesicle trafficking and membrane link domain containing 1)is a candidate cancer suppressor gene discovered in our laboratory,its expression is significantly downregulated in various cancer tissues such as breast cancer,lung cancer,etc.The mechanism of its function has not been ascertained yet.In the previous study,the results showed that MarvelD1 is mainly located in the nucleus and mitochondria.Mitochondria is the main organelle producing the endogenous reactive oxygen species(ROS).The imbalance of the electron transport chain in mitochondria leads to the leakage of ROS.When the production and clevage of ROS in the body under an imbalance situation,the body will be in an oxidative stress(OS)state.DNA and protein could be damaged under OS state.OS is often associated with diabetes,cancers,and various diseases.Metabolomics is a newly emerging subjest belongs to the system biology.Using metabolomics methods to study organisms under OS can amplify the alternation in genes expression,thereby it could observe the body's metabolism more sensitive.Therefore,using metabolomics methods to explore the function of MarvelD1 in OS can provide data support for the next research.In this report,the wild-type mouse C57BL/6J and the MarvelD1 whole-body knockout mouse were used as a model animal.The mice were fed with 1500 ppm potassium bromate water to construct an OS model and then we performed ELISA to detect the biomarker of OS.At the same time,mice were metabolic monitoring at the young,middle and old age.Utilizing the UPLC-QTOF/MS platform,we collected a total of 105 mice sera and obtained its metabolic profile.Principal component analysis(PCA)was conducted to analyze the metabolic profile between different samples.The differential ionization was screened with p<0.05,VIP>1 and performed secondary identification.The results demonstrated that there was a significant difference in serum metabolism between MarvelD1-knockout mice and wild-type mice.Therefore,the differential metabolites were mainly sphingolipid and carbohydrates.And there is a significant change in amino acids metabolism between the OS model mice and the normal mice.Metabolic analysis was performed by using the Metabo Analyst website after obtaining differential metabolites between multiple groups.The results showed that the MarvelD1-knockout mice showed significant imbalance in the sphingosine metabolic pathway compared with the wild-type mice.There are also significant changes in phenylalanine metabolism and genesis pathway and carnitine metabolism pathway between OS model mice and normal mice.Further studies revealed that when the MarvelD1-knockout mice undergo the OS,their pentose phosphate metabolic pathway(PPP)has a significant downward trend,but this trend is not shown when wild-type mice undergo the OS or MarvelD1-knockout mice with normal feeding conditions.The rate-limiting enzyme in the PPP is G6 PD.Mouse fibroblast cell MEF was used to construct a MarvelD1 knock-out model cell,and H2O2 was used as an inducer of OS.The cells are treated 24 h,48h and 72 h with H2O2 and detect the G6 PD expression by using QPCR.The conclusion is totally the same as the serum analysis.This shows that MarvelD1 may regulate the mouse's PPP pathway by affecting the expression of G6 PD,leading to the significant changes in MarvelD1-knockout mice under the OS state.In this study,an established UPLC-QTOF/MS analysis platform was used to analyze mice serum metabolic profile under different ages,gene type and oxidavice stress state.Differential metabolites that characterize the differences between groups were analyzed for metabolic pathways and were verified by cytology.The results of this study provide further insights to the function research of MarvelD1.