| Purpose:By comparing the methylation status of imprinted gene H19 in three different quality spermatozoa and the methylation status of the embryos after the combination of spermatozoa,the sperm methylation status in the ART technique was preliminarily explored with the postnatal embryo methylation And to provide a theoretical basis for ART to reduce the risk of epigenetic disease.Methods:In this study,a total of 91 spermatozoa and 91 low-quality embryos were collected from IVF-ET in the First People's Hospital of Yunnan Province.Among them,sperm samples were divided into three groups according to the WHO fourth edition sperm concentration parameter Group A,control group,normal sperm parameters group,sperm concentration ? 15 X106/ml,a+b?32%,44;B group,oligospermia(including mild oligospermia/severe oligospermia)group,sperm concentration<15 X106/ml,a+b<32%,26;C,testis/epididymal sperm group,21,group B and group C were experimental group.The DNA methylation patterns of the H19 and PEG1 DMR regions in sperm and embryos were determined by the method of Sulfite sequencing PCR(BSP),and the relationship between different quality spermatozoa and postnatal embryo methylation was determined.Result:(1)Imprinted gene H19 Methylation status:a:10 samples of methylated abnormalities(11.0%,10/91)were found in 91 sperm samples.There were 6(23.1%,6/26)methylation abnormalities in group B.There were 1(4.8%,1/21)abnormalities in group C,and methylation abnormalities in group A 3(7.5%,3/40).(Pab = 0.040;Pbc = 0.007;P = 0.014)in group A and group B,group B and group C,group A and group B.group C were significantly different Group was not statistically significant(P>0.05).b:A total of seven abnormal low methylation patterns were found in 55 successful embryos(12.7%,7/55),among which there were 5 belonging to group B and group C(17.2%,5/29),There were 2 in Group A(7.7%,2/26),and there was significant difference between the two groups(P = 0.040).2.7%methylation status of normal sperm fertilization after embryonic abnormalities;10%abnormal methylated sperm corresponding to the embryo did not appear abnormal methylation status;IVF group and ICSI group chi-square test was not significant(P>0.05).(2)imprinted gene PEG1 methylation status:a:13 samples of methylated abnormalities were found in 91 sperm samples(14.4%,13/91).There were 8(26.7%,8/30)methylation abnormalities in group B,and 3(14.3%,3/21)in group C,and abnormal methylation status in group A There are 2(5.0%,2/40).The difference between the groups was statistically significant:chi-square test:Group A and Group B,Group B and Group C,Group A and B.C group methylation abnormalities were significantly different(Pab = 0.006,Pbc ? 0.010,P = 0.003).There was no significant difference between group A and group C(P>0.05).b:A total of 11 embryos were found to be abnormal hypermethylation patterns(21.2%,11/52)in 52 successful amplified embryos.Among them,10(43.5%,10/23)belonged to group B and group C,belonging to group A There were 1(3.4%,1/29)and chi-square differences in the two groups(P = 0.010).There was no significant difference between the IVF group and the ICSI group(P>0.05)Conclusions:1.The abnormalities of H19 and PEG1 methylation were mainly concentrated in the abnormal parameters of spermatozoa,suggesting that the abnormal state was related to the decrease of sperm quality,and there were few weaker sperm diseases with genetic defects,and the patients with azoospermia Pass to the offspring;2.Through IVF,ICSI fertilized embryos have the same proportion of the corresponding abnormal state,no significant difference.3.abnormal sperm embryos did not appear abnormal,which may be related to the self-repair function of the embryo itself;and individual normal sperm fertilization after embryonic abnormalities of methylation may be related to the conditions of in vitro culture,but also possible with the egg quality Its own;... |
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