| Type 1 diabetes mellitus,morbility age is small,for women of childbearing age,niche targeting therapy especially control of blood glucose is very important for mother and fetus.Diabetes Mellitus affect fetus through many ways,among them,the influence of blood glucose may be a key factor.The macrosomia caused by high level of glucose has already been reported.Therefore,under insulin therapy condition,the low level of glucose may injure the fetus also.The fetus of GMD mother's mutation rate is two or three times than healthy ones.During the early period of pregnancy,especially on embroic day 6 the organgenesis begin,if fetus being affected by unstable enviroment fetus will get mutation.Meanwhile,the fetus from Gestational diabetes mellitus mother whose blood glucose are unstablely controlled,their accuracy movement are weak,they may suffer from movement dysfunction and unconcertration.Clinical work has reported that the fetus from Gestational diabetes mellitus mother,prat of them has retard brain development.Therefore the early effect of Gestational diabetes mellitus effect on child has rarely reported.In our study we use Streptozotocin to induce Gestational diabetes mellitus.Our aim is to syudy the realationship in embroic day 14,16,18,and the change of insulin and insulin like growth factor1 in feyus brain.Methods:Wistar f-emale rats(? 12,body weight 250g-260g),Wistar male rats((?)12,body weight 250g-260g),obtained from the experimental animal center of Lanzhou University.Twelve female Wistar rats and twelve male Wistar rats are randomly divided into two groups respectively.After 12 hours light 12 days dark cycle for one week,female rat and male rat mate at night.The nextday morning observe the vaginal smear slide,if founding spermatid,noting pregnancy positive one day.At the pregnancy day 2,65mg/kg Steptozotocin injected inraperitoneally(STZ induced group).Analyze blood glucose on 8:00am 14:00 pm 20:00pm until blood glucose increas 3 times than normal level(>350 mg/dL).And then 20/U insulin hypudermic injected everyday until the pregynancy day 14 16 18.(STZ + Insulin group)..At the pregnancy day 14 1618,female rat is going to anesthetize with Chloral Hydrate by intraperitoneal injection.The fetus get cut from uterine.And then fetus' brain stored in liquid nitrogen.All steps is operated under low temperature.Indexes detected fetal rat neuron spines' density analysis:HE staining make right slice,use golgi-cox staning to observe.Analyze synaptoshysin:using western-blot to analyze the expression of synaptoshysin.The expression of Insulin,insulin like growth factor-1 and their receptor:agarode gel electrophoresis and PT-PCR analyze Insulin,insulin like growth f-actor-1 and their receptor gene expression.Results:1.The effect of blood glucose consentration caused by insulin treatment in gestational rat.The insulin treatment gestational rat's blood glucose concentration maintain at 10 mol/L,The non-insulin treatment gestational rat' blood glucose concentration maintain at 22 mol/L,thus normal rat's blood glucose concentration maintain at 6 mol/L.2.The effect of neuron spines density in fetal cortex caused by high level of blood glucose concentration.Under high level of blood glucose concentration The fetus's neuron spines density is 5.4/mm2 developed at embroic day 14.at embroic day 16 is 7.2/mm2,at embroic day 18 is 9.8/mm2.Under insulin treatment The fetus's neuron spines density is 9.6/mm2 developed at embroic day 14.at embroic day 16 is 14.1/mm2,at embroic day 18 is 22.2/mm2.3.The change of synaptic marked protein SYN in fetus's brain under high level of blood glucose concentration.Under high level of blood glucose concentration the expression of Synaptophasin is about 55%compare to insulin treatment group at embroic day 14,at embroic day 16 is about 50%,embroic day 18 is about 42%,4.The change of insulin and insulin like growth factor 1 mRNA in fetus's brain under high level of blood glucose concentration.Under high level of blood glucose concentration the expression of insulin is about 74%compare to insulin treatment group at embroic day 14,at embroic day 16 is about 63%,embroic day 18 is about 52%,Under high level of blood glucose concentration the expression of insulin like growth factor 1 mRNA is about 122%compare to insulin treatment group at embroic day 14,at embroic day 16 is about 130%,embroic day 18 is about 146%,5.The change of insulin and insulin like growth factor 1 receptor mRNA in fetus's brain under high level of blood glucose concentration.Under high level of blood glucose concentration the expression of insulin receptor mRNA is about 116%compare to insulin treatment group at embroic day 14,at embroic day 16 is about 130%,embroic day 18 is about 142%,Under high level of blood glucose concentration the expression of insulin like growth factor 1 receptor mRNA is about 118%compare to insulin treatment group at embroic day 14,at embroic day 16 is about 124%,embroic day 18 is about 134%,6.The change of insulin and insulin like growth factor 1 plus it's receptor protein in fetus's brain under high level of blood glucose concentration.Under high level of blood glucose concentration the expression of insulin receptor protein is about 120%compare to insulin treatment group at embroic day 14,at embroic day 16 is about 131%,embroic day 18 is about 142%,Under high level of blood glucose concentration the expression of insulin like growth factor 1 protein is about 117%compare to insulin treatment group at embroic day 14,at embroic day 16 is about 129%,embroic day 18 is about 144%,Under high level of blood glucose concentration the expression of insulin like growth factor 1 receptor protein is about 122%compare to insulin treatment group at embroic day 14,at embroic day 16 is about 131%,embroic day 18 is about 142%,Conclutions:1.Gastational Diabete mellitus rat's blood glocuse consentration can be proximitly controlled as normal level by long acting insulin injection.2.The fetal rat's brain insulin level decreased caused by maintaining high blood glocuse consentration,but fetal rat's brain Insulin like growth factor-1 level increased conpensatorily.3.Under high blood glocuse consentration,the expression of Insulin/Insulin like growth factor-1 receptor increased.4.The fetal rat's brain Synaptophysin level decreased caused by maintaining high blood glocuse consentration,meanwhile the density of neuron spines decrased also. |
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