| Gestational diabetes mellitus(GDM)is a common pregnancy complication that can cause adverse pregnancy outcomes such as giant babies,postpartum haemorrhage and increased caesarean section rates,and is closely associated with the development of distant adult disease in the offspring.The pathogenesis of GDM is not yet clear,and recent studies have identified intestinal microecological imbalances that may be involved in its pathogenesis.ObjectiveTo investigate the relationship between the feeding factor Nesfatin-1 and intestinal flora and its involvement in the pathogenesis of GDM by analyzing the diversity and abundance changes of intestinal flora in a maternal population with different body mass index(BMI)before pregnancy by 16 sr RNA high-throughput sequencing,in order to guide the early prevention and targeted treatment of clinical GDM.MethodsA total of 33 pregnant women diagnosed with GDM in the obstetric outpatient clinic of Qingdao Municipal Hospital from July 2021 to October 2022 were included in the study who met the criteria.They were divided into normal weight group A(20 cases)and overweight/obese group B(13 cases)according to their pre-pregnancy BMI.Maternal serum was collected from 24 to 28 weeks and stool samples were collected at the same time.(1)Serum Nesfatin-1 level was measured by enzyme-linked immunosorbent assay(Elisa),glycosylated haemoglobin(Hb A1C)by high performance liquid chromatography ion exchange,blood insulin by electrochemiluminescence,and serum total cholesterol(TC)and triglyceride(TG)by enzyme-linked endpoint assay,enzymatic assay for the detection of serum bile acids.(2)The intra-and inter-group diversity,significant species differences and correlation analysis of faecal intestinal flora were analysed by 16 sr RNA high-throughput sequencing and bioinformatics.(3)SPSS 22.0 and R v3.1.1 software were used to analyse and process all data.Data on measures were expressed as ?x±S,and means between two groups were compared by t-test;correlation analysis was performed by Pearson analysis.p<0.05 was considered a statistically significant difference.Results(1)The level of Nesfatin-1 expression was reduced and the levels of Hb A1 C,TC and TG were increased in group B compared with group A.The differences were statistically significant(P<0.001).The differences between the two groups in the magnitude of weight gain,fasting insulin and serum bile acids were not statistically significant.(2)In all patients,serum Nesfatin-1 levels were negatively correlated with BMI,fasting insulin,Hb A1 C,TC and TG,and positively correlated with serum bile acids,with statistically significant differences(P < 0.05);there was no significant correlation with the number of weight gain.(3)Structural analysis of the intestinal flora of the two groups showed that there were significant differences in species abundance and species diversity between the two groups at all levels.At the phylum level,the common phylum detected in both groups were Bacteriodetes,Firmicutes,Proteobacteria,Verrucomicrobia and Actinobacteria.The abundance of the phylum Verrucomicrobia was higher and that of the phylum Actinobacteria was lower in group B compared with group A(P < 0.05).The phylum Verrucomicrobiae,Clostridia,Bacteroidia,Negativicutes and Gammaproteobacteria were common to both groups at the phylum level.The abundance of B-group was lower than that of A-group,while that of B-group was higher than that of A-group(P < 0.05).At the order level,Oscillospirales,Bacteroidales,Bacteroidales and Peptococcales were the most common orders in the two groups,with Clostridia and Bacteroidales being more abundant in group A and Peptococcales in group B(P < 0.05).The common families in both groups at the family level were Akkermansiaceae,Clostridia,Rikenellaceae,and Ruminococcaceae.The abundance of Akkermansiaceae,Clostridia,and Rikenellaceae was lower in group B compared to group A(P < 0.05).At the genus level,the bacteria detected in both groups were Akkermansia,Fournierella,Ruminococcaceae and Bacteroides,etc.Compared with group A,the abundance of Akkermansia,Bacteroides,Fournierella and Ruminococcaceae was lower in group B The abundance of Ackermannia,Bacteroides,Fournierella and A.tumefaciens was lower and that of Sellimonas was higher(P < 0.05).At the species level,Ackermannia spp.and Fournierella spp.were less abundant and UCG＿010 was more abundant in group B compared with group A(P < 0.05).(4)The alpha diversity analysis by Chao1 index and Shannon index showed no statistically significant difference in the diversity of intestinal flora between the two groups(P=0.38),while the difference in abundance was statistically significant(P=0.00011).(5)Nesfatin-1 was strongly correlated with Ackermannia,TC,TG and E.faecalis spp.by RDA analysis and was positively correlated.(6)Pearson correlation analysis of serum Nesfatin-1 and lipid index with intestinal flora diversity showed that Nesfatin-1 was positively correlated with intestinal flora diversity(r=0.631,P<0.001),TC and TG were negatively correlated with intestinal flora diversity(r=-0.601,P<0.023),(r=-0.642,P = 0.018).Conclusions1.In pregnant women with GDM,peripheral blood Nesfatin-1 and glucolipid metabolism vary according to BMI.2.Flora diversity in pregnant women with GDM was not altered by pre-pregnancy BMI,but flora abundance was reduced in overweight/obese individuals.3.In pregnant women with GDM,the relationship between peripheral blood Nesfatin-1 and Ackermannia may provide clues to the mechanisms by which intestinal flora regulate feeding.4.In pregnant women with GDM,intestinal flora diversity may positively regulate peripheral blood Nesfatin-1 to influence feeding.5.The feeding factor Nesfatin-1 and intestinal flora may be related through the regulation of glucolipid metabolism in order to influence the development of GDM. |
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