| Objective:This subject will discuss Zuogui pill of Chinese medicine’s effect to IGT’s susceptibility gene expression and methylated modification from the aspect of regulate Epigenetics by observe the pill’s preventing effect in intervening IGT of the second generation rats which born in intrauterine’s high glucose environment in embryonic period, and inspect the effect to pregnant rats’s embryo’s function.Methods:Sorting pregnant rats in 5 groups randomly, which are control group (C)> intrauterine’s high glucose group (M), Zuogui pill of Chinese medicine group (ZGG), insulin intervene group (INS), and combined intervened group (F). At the day of pregnant, except the control group, the other 4 groups are intraperitoneal injected by streptozotocin (45mg·kg-1) to copy intrauterine’s high glucose model. At the 3rd day of pregnant, given rats Zuogui pill of Chinese medicine group or insulin as intervene methods untill the 19th day. Make part of pregnant rats into death at the 19th day of pregnancy, test the bood glucose, insulin and corticosterone of pregnat rats with ELISA. Detect mRNA and protein expression level of 11(3-HSD-2 inplacenta with RT-PCR and Western blot technique. After every pregnant rats give birth to their children, Every secondary rats are feed by mother rats for 3 weeks and regular fodder for 2 weeks. From the 6th week, except the control group is feed by regular fodder continuously, other 4 groups are feed by high-glucose and high-fat fodder. Every week, test the rats with Glucose tolerance test, untill the ntrauterine’s high glucose group appear IGT, then we make all groups’ rats to death. (1) Observe the birth weight and growth curves of every secondary rats. Test their glucose and blood lipid in empty stomach with automatic biochemical analyzer.test insulin and adiponectin with ELISA. Observe liver’s and pancreas’s pathology morphological characteristics which dyed by HE under microscopic, and the micro structure of pancreas’s cell under electron microscopy. (2) Test the mRNA of APN which is correlated with pancreas cell’s growth and IGT and diarrhea’s pathogen with PCR. Test the methylation state of APN with bisulfite genomie sequencing.Results:1.The birth weight, glucose-fat metabolism, pathogen change of liver and pancreas in intrauterine’s high glucose group is abnormal, and these factors can be differently prevented and treated by drug intervene. Among these drugs, the combination of Zuogui pill of Chinese medicine and insulin has the best effect.2.APN’s mRNA expression in fat of the secondary rats’intrauterine’s high glucose group is lower than control group. APN’s mRNA expression in fat of combination drug intervened groups has no significantly difference with control group. APN expression in drug intervened groups are inverse with their DNA’s methylation. 3.Expression of 11β-HSD2’s mRNA in drug intervened group is significantly higher than intrauterine’s high glucose group. In the drug intervened groups, corticosterone in blood decreased, secondary rats’weight increased, and they have no difference between combined intervened group and control group.Conclusions:The combination of Zuogui pill of Chinese medicine group and insulinin intervene in embryo period can prevent the pathogen to secondary rats’IGT when it is in intrauterine’s high glucose environment, and its mechanism may as follows:1.Intervene in embryo period can control the disease from very beginning, by regulating expression of embryo’s 11β-HSD2, cutting the influence of bad environment in the placenta, lightening growth sluggish in placenta, decrease the risk of IGT’s pathogen in adult.2.The intervene in embryo period can effect APN’s mRNA expression and methylation state, so that it prevent the abnormal metabolism.
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