| Research BackgroundDiabetes(Diabetes Mellitus,DM),a refractory,systemic metabolic disease,seriously affects people’s quality of life.Human society has an urgent need to overcome the issue.Pregnant women with obesity,gestational diabetes and various maternal excess nutrients in the uterine environment have a significant impact on future health problems,and their offspring are at high risk of impaired diabetes and sugar regulation.Chinese medicine theory"kidney is innate,the main reproduction,possession of fine" and "acquired embryo quality" is closely related.Chinese medicine believes that human growth and development,breeding offspring with its kidney gas generation,strong,weak and closely related.Congenital affect the day after tomorrow.Kidney is the congenital of this,the main reproduction,possession of fine.Kidney essence is sufficient,strong fertility;kidney essence deficiency,reproductive capacity is weak.Kidney does not charge,eggs,sperm,fertilized eggs,embryo quality decline.Zuogui Pills Peiyuan Yuan Yin,leaving the blood self-sufficiency,can "make up the kidney essence,the benefit of the future embryo."Previous experiments show that Zuogui pill not only reduce the fasting blood glucose in diabetic rats,but also reduce the gestational diabetes maternal triglyceride,total cholesterol,insulin levels,while the pancreatic islet B cells play a protective role in gestational diabetes(IGT)induced by high glucose and high fat diet in the offspring of GDM rats.Zuogui pill containing serum in vitro and in vivo role similar to the resistance to alcohol inhibition,and promote the development of preimplantation embryos in vitro.Therefore,the appropriate high glucose-loaded mouse preimplantation embryo development model was used to observe the effect of Zuogui pill containing serum.Purpose of researchTo analyze the migratory chemical constituents of Zuogui pill containing serum,and to study the influx of blood in Zuogui pill.To observe the effect of high glucose on the in vitro development of preimplantation embryos in mice,and to observe the effect of Zuogui pill serum on the in vitro development of preimplantation embryos in high glucose rats.Research methodsThe origin and structure of the migratory components in serum were analyzed by UPLC-Q/TOF-MS chromatogram.The elemental composition of Zuogui pill was analyzed by OPLS-DA analysis.Into the blood component.In vitro culture of ICR mouse embryos,fertilization 38h when the statistics of 2-cell rate,fertilization 110h when the blastibility rate.The glucose content was gradually increased to 80mmol/L(blank group 2.81mmol/L),divided into 5 groups:blank group,20mmol/L glucose group,40mmol/L glucose group,60mmol/L glucose group,80mmol/L glucose group.All the fertilized eggs collected in one experiment were randomly divided into two groups.The effects of high glucose on preimplantation embryo development were observed.The concentration of 40mmol/L was used as a high glucose to inhibit the in vitro development of preimplantation embryos in vitro.The serum containing Zuogui pill was used to investigate the effects of high glucose on the in vitro development of preimplantation embryos in vitro.Embryo rate and blastocyst cell count were used to evaluate the quality of embryos.The effects of Zuogui Pill containing serum on the in vitro development of preimplantation embryos in high glucose rats were observed.Research resultUPLC-Q/TOF-MS chromatographic analysis showed that 26 influx components were detected in negative ion mode,including 12 prototype components and 15 metabolites.In the negative ion mode,13 blood influencing factors were identified,among which were the species of malt glycosides,loganoside,monosine,malarginine,epimerosine derived from dogwood,and 5 Hydroxymethyl-2-furfural glucuronic acid glycoside,dihydro-5-hydroxymethyl-2-furfural glucuronide,diclofenac,vanillic acid derived from medlar,kaempferol derived from dodder Aldehyde glycosides.2-cell rate was most closely related to fertilization,and the high glucose environment had no effect on 2-cell,while the blastocyst rate decreased with the increase of glucose concentration.The blastocyst rate in the blank group was above 70%.Mostly to 3,4 blastocysts,showing a small amount of 5 blastocysts.20mmol/L glucose group was similar to that of blank group,and there was no obvious difference in morphology.There was no significant difference between the two groups.The blastocyst rate of 40 mmol/L glucose group decreased in 2 stage,3 Period,showing 4 blastocysts,blastocyst morphological changes significantly,the inner cell group thick and close,small cysts cavity,cavity fluid less,in the early stage of embryos accounted for the majority of embryos.The blastocyst rate of the 60 mmol/L glucose group was significantly decreased(p<0.01),and the blastocysts of 40 mmol/L glucose group were similar in 2 and 3 stages.The blastocysts were in the early stage and the blastocysts were small.80mmol/L glucose group almost no blastocysts,embryos ranging from 2 to 4 cells,most embryos were blocked in the 2-cell period.With the increase of glucose concentration,the number of bad eggs in the culture droplets also increased accordingly.The rate of blastocysts decreased with the increase of sugar concentration.The number of early blastocysts increased in the total blastocyst number(including early blastocysts,blastocysts and hatching blastocysts).When the sugar concentration increased to 80mmol/L,The formation is completely suppressed.It is reasonable and effective to use 40mmol/L glucose to inhibit embryonic development as a model for the intervention of embryos.Zuogui pill serum can resist high glucose inhibition,promote the suppression of embryonic development.From the blastocyst rate,Zuogui pill containing serum can effectively promote embryonic development,and can eliminate the impact of blank serum.The number of blastocysts in the Zuogui pill group with higher blastocyst rate was less than that in the blank group.in conclusion1.Zuogui pill serum is mainly enriched iridoid glycoside active ingredients,witch is an important basis for the efficacy of substances.2.The parental high glucose environment inhibits the development of preimplantation embryos.It is reasonable and effective to use 40mmol/L glucose to inhibit embryonic development as a model for the intervention of embryos.3.Ziyin Bushen prescription Zuogui pill fights against the high sugar inhibition,promote embryonic development. |
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