| Objective:Recurrent spontaneous abortion(RSA)is one common clinical pregnancy complications in women of childbearing age.In recent years,research has shown that the incidence of RSA has a tendency to rise.The etiology of RSA is complicated,which involves genetic,the structure of the genital tract,endocrine,immune and other aspects.With the development of molecular biology,people began to pay attention to the function of miRNA and Leptin on the disease.So far,there are few studies reported the relationship between Leptin and infertility,and there were less research about the relationship between RSA and the gene polymorphism of miRNA-27a and Leptin.Thus,our study intends to explore the impact of gene polymorphisms of miRNA-27a and Leptin on RSA,and evaluate the regulatory function of miRNA in the process of pregnancy,and discuss the effect and significance of the miRNA-27a and Leptin on RSA.Methods:Between April 2013 and April 2015 at Shenzhen longhua New District Central Hospital,we randomly recruited 138 RSA patients as case group.And another 142 normal pregnancy women were included as control group.Take peripheral blood(5 ml)from each subject on an empty stomach.The real-time quantitative polymerase chain reaction(RTQ-PCR)was used to measure the level of miRNA-27a.The Enzyme-linked Immunosorbent Assay(ELISA)was used to measure the level of Leptin.The denaturing high performance liquid chromatography(DHPLC)was employed to determine the genotypes and allele frequencies of miRNA-27a rs895819 A/G and Leptin rs7799039 G/A.Continuous data were expressed as mean±standard deviation(SD),with t test for comparison.Categorical data were exhibited as percentage or rate,with x2 test for comparison.The χ2 test was also employed to identify whether samples between two groups met the Hardy-Weinberg equilibrium.Besides,odds ratio(OR)with 95%confidence interval(CI)was calculated to estimate association between disease and polymorphism in the single-factor and multi-factor logistic regression analyses.Results:1.The test of distributions of genotype and allele frequencies of miRNA-27a rs895819 A/G and Leptin rs7799039 G/A in RSA group and control group is consistent with Hardy-Weinberg equilibrium,which demonstrated that the study population was representative.2.With respect to miRNA-27a rs895819 A/G,the frequencies of GG genotype and G allele were both significantly higher in the case group than the control group.As for Leptin rs7799039 G/A,the frequencies of AA genotype and A allele were also both higher in the case group than the control group.3.The polymorphisms of miRNA-27a rs895819 A/G and Leptin rs7799039 G/A could increase the risk of RSA.4.Compared with individuals with AA-GG combined genotypes,GG-AA and AG-AA carriers had an increased risk of RSA.5.There were significant differences in clinical features between patients with G allele and non-G allele in miRNA-27a rs895819 A/G.There were also significant differences in clinical features between patients with A allele and non-A allele in Leptin rs7799039 G/A.6.MiRNA-27a expressed in the patients with AA genotype and AG+GG genotype of rs895819 A/G as well as Plasma Leptin level of patients with GG genotype and GA+AA genotype rs7799039 G/A in both case group and control group,with an significant difference in its total expression volume between the control group and case group.Conclusion:1.The mutation of G gene of miRNA-27a rs895819 A/G and mutation of gene A of Leptin rs7799039 G/A might increase the risk of RSA.2.Compared with individual with combined AA-GG genotype distributed on miRNA-27a rs895819 A/G locus and Leptin rs7799039 G/A locus,patients with combined genotype of GG-AA and AG-AA suffered a higher risk of RSA... |
