GC-MS Was Used To Detect The Effect Of Zuogui Pill-containing Serum On The Embryonic Development Damage Of High Glucose Mic

Objective:Study on high concentration glucose add to human tubal fluid of ICR mouse embryos(zygote to blastocyst)affect the development of in vitro and adding serum intervention on the impact of the role.Based on the comparative analysis of the metabolites in the blastocyst stage,the possible mechanism of the action of the Zuoguiwan was discussed.Methods:HTF medium containing 0mmol/L,20mmol/L,40mmol/L,60mmol/L glucose was used,ICR mouse embryo cells were cultured in vitro by m icro drop method and CO2 incubator 120h,The changes of 2-cell rate(24h),blastocyst rate(120h)and number of blastocysts(120h)in different glucose concentrations were observed by stereo microscopy and fluorescence staining.Selectting the suitable glucose concentration which could inhibit the early embryonic development of ICR mice at th e same time,add 5%or 10%left pill serum to act on the 1-cells in vitro,and observe the changes of 2-cell rate,blastocyst rate and the number of blastocysts.GC-MS,OPLS-DA model and hierarchical clustering were used to analyze the metabolic compounds of high glucose blastocy st stage and high glucose blastocyst stage.Results:Embryonic cells can incubate and develop normally in the HTF medium added with 0mmol/L and 20mmol/L glucose,In HTF medium containing 40,60mmol/L glucose,development was significantly impaired(P<0.05).The concentration is directly proportional to the damage.The serum containing 5%or 10%left pills could reduce the damage of HIF medium containing 40 and 60mmol/L glucose on embryonic development(P<0.05).Using gas chromatography GC-MS detection tech nique,metabolic group results showed:High glucose groups of phenylalanine,succinate,glucose,-1-phosphate,D-glucosamine,-1phosphate,and 6-deoxy-D-glucose levels were up-regulated,whereas uracil,glycine,and citrate levels were down regulated.The levels of lin oleic acid and glycine in the serum group were increased,mannose,arachidonic acid,and the content of four amino acids decreased,Differences in metabolic pathways involved in carbohydrate metabolism pathways(including glucose metabolism of starch and sucrose metabolism,galactose);carbohydrate metabolism(citrate cycle);amino acid metabolism pathway(phenylalanine biosynthesis,metabolism,phenylalanine,tyrosine and tryptophan glycine,serine and threonine metabolism,beta alanine metabolism);lipid metabolism pathway(glycerol metabolism,linoleic acid metabolism).Conclusion:It suggested that the serum contained in the left pill was helpful to improve the em bryonic development arrest caused by high glucose load.It has the effect of correcting the embryonic development inhibition caused by high concentration of glucose in cell morphology.It indicated that the embryo development damage caused by the high concentration of glucose in the left pills has more than the morphological repair effect.High glucose may cause fetal abnormal energy metabolism,the final metabolic pathway-citrate cycle abnormal protein and DNA lack of raw materials,abnormal brain development,oxidative ability.Zuogui pill has antioxidant,regulating immune response and reduce the immune response to injury,added amino acids,repairing brain injury.Embryo dam age on glucose induced with at the same time for the repair effect of having enhanced resistance to withstand the other injury.