The Study Of Urine Metabolomics Of RSA Rats By GC-MS Method

ObjectiveBased on the GC-MS method,the study on the effect of Shou Shou Pill on the urine metabonomics of rats with recurrent spontaneous abortion,Describe the specific mechanism of the operation of Shou Tai Wan.Method1.The spontaneous abortion model and the establishment of normal pregnancy model:SPF grade SD rats,conventional observation of 1 week,40 female rats were 20 male rats by 2:1 cage mating,check the thrombosis Counted as the first 0 days of pregnancy,respectively,the establishment of repeated spontaneous abortion model 30 and normal pregnancy model 10.2.Grouping and administration of repeated spontaneous abortion rat models:Female pregnant mice were randomly divided into 4 groups:ShouTaiWan group,progesterone group,model group and normal pregnancy group.Female rats were treated with vaginal smear on the second day of the morning.Under the microscope,a large number of spermatozoa were found in the vaginal smear.The pregnant rats were divided into gestation day 0,ShouTai pill group,progesterone group and model group Pregnancy 1 day to 10 days to hydroxyurea(450mg.Kg-1,saline configuration)gavage,the first 10 days of pregnancy to mifepristone(3.75rag.Kg-1,saline configuration)gavage,life(8ml.Kg-1.d-1)for 10 days;the progesterone group from the first day of pregnancy,given progesterone intramuscular injection(the first day of pregnancy)8mg.Kg-1)for two days,until the 10th day of pregnancy;model group at the same time to give the same amount of saline saline for 10 days,the normal pregnancy group from the first 0 days of pregnancy given saline(8 ml.Kg-1.d-1)for 11 consecutive days.At the end of treatment,the rats were collected and the urine was collected and the rats were sacrificed.The urine samples were stored in a-20 degree Celsius refrigerator,and a small amount of urine was subjected to batch pretreatment.A 1.0 ?L sample was used for GC-MS analysis.Result:1.This experiment identified 82 species of effective differential metabolites.2.Shoutai Pill group and the model group metabolic profiling of urine significant changes,2-methyl-Butanedioic,Serine,Alanine,Glucaric acid,Quinic acid,Pyroglutamic acid,Gentibiose,Threonine,Phenylalanine,Isoleucine,Uridine,Arabitol,Valine,Tyrosine,Ribitol,Threitol,Citramalic Acid,pantothenic acid in the urine Shoutai pill group increase significantly the level(P<0.05),and uracil,2-ketoglutaric acid,uric acid,Lactic acid,Heptanoic acid,3-Hydroxypyridine,Adipic acid,Succinic acid,Glyceric acid-3-phosphate,Shou Tai Wan group of urine levels were significant Reduction(P<0.05),these metabolites is raised and lowered Shoutai abortion pill certain correlation.The levels of urinary metabolism in the progesterone group were significantly different from those in the model group.Threonine,Hydroxylamine,Pyroglutamic acid,Hexonic acid,Gentibiose in the progesterone group(P<0.05),while 2-Ketoglutaric acid,Lactic acid,Glyceric acid,3-Hydroxypyridine,Uric acid,uracil,glycolic acid,heptanoic acid,adipic acid in the urine in the progesterone group decreased significantly(P<0.05),which The difference between the two groups of metabolites was associated with progesterone.There was also a significant change in urine metabolic profile and model group in normal pregnancy group.The levels of Glyceric acid-3-phosphate,phosphoric acid and glycerol were significantly increased in the urine of normal pregnancy group P<0.05),and serine,cysteine and other 48 kinds of metabolites in urine levels decreased significantly in normal pregnancy group(P<0.05),compared to normal pregnancy groups and the model group It can be seen This is some of the differential metabolites associated with miscarriage.Conclusion1.There was a significant difference in metabolic profile between the model group and the ShouTaiWan group,the model group and the progesterone group,and the model group and normal pregnancy group.we will be in the future clinical work for further study.2.Gas chromatography technology revealed detailed information on the metabolic profile of the urine in the model group,ShouTaiWan group,progesterone group and normal pregnancy group,thus demonstrating the feasibility of the method.At the same time,PCA model can determine the metabolic difference of urine in ShouTaiWan group.In addition,82 metabolites were screened out.Based on this trial,we believe that this approach will play a vital role in the treatment of recurrent spontaneous abortion.3.This experiment shows that Shou Touwan mainly through the body of phenylalanine,tyrosine,valine,isoleucine,pyroglutamic acid,affecting catecholamines,thyroxine,glutamine synthesis to make tires The effect.However,Shou Tou Wan treatment of recurrent miscarriage of the clear mechanism of action need to do in-depth study.