| ObjectiveTo investigate the differences in the composition and abundance of gut microbiota in children with short stature after recombinant human growth hormone(GH)treatment by 16S rDNA high-throughput sequencing and bioinformatics analysis,and to analyze the correlation between gut microbiota and growth-related indexes,with the aim of providing new ideas and treatment options for improving the growth of children with short stature in the future.MethodsThere were fifty children aged 3 to 14 years old who first treated with GH from July 2021 to June 2022 in Subei Hospital were selected and divided into long-acting group(Group L)and short-acting group(Group S)according to the dosage form of GH used.In the Group L,Polyethylen Glycol Recombinant Human Growth Hormone(PEG-rhGH)was used,and Group S,Recombinant Human Growth Hormone(rhGH)was used.Liver and kidney function,thyroid function,and glycosylated hemoglobin were monitored regularly during treatment with GH.Fresh stool specimens and clinical data were collected and recorded,at the beginning,3 months and 6 months of treatment,and further grouped into L0,L3,L6,S0,S3 and S6 according to the follow-up time.The correlations between gut microbiota and growth velocity(GV),standard deviation score(SDS),and insulin-like growth factor-1(IGF-1)were analyzed.Results1.There were no statistically significant differences in gender and age between the Group L and Group S(P>0.05),and no significant abnormalities in liver and kidney function,thyroid function,or glycated hemoglobin were observed in the children monitored during GH treatment.At 3 months of treatment,IGF-1 levels were significant higher in Group L3 than those in Group S3(213.22 ± 80.97 VS 174.01 ± 52.76 ng/ml,t=4.400,P=0.000).At 6 months of treatment,GVs were significant faster in Group L6 than those in Group S6(1.10 ± 0.30 VS 0.82±0.19 cm/m,t=3.039,P=0.006)and height SDS were significant better in Group L6 than those in Group S6(-1.27±0.09 VS-1.45 ± 0.12,t=3.607,P=0.002).2.The relative abundance and composition of gut microbiota species showed that the abundance of Bacteroidota and Bacteroidetes in both L and S groups increased gradually after GH using,and the ratio of Bacteroidota/Firmicutes(B/F)also increased gradually,while the abundance of the rest of the flora fluctuating changed,but the differences were not statistically significant(P>0.05).3.The alpha diversity indices(Chao1,Pielou e indices)of the L and S groups gradually increased with the prolongation of GH using,and the richness and homogeneity of the gut microbiota were continuously improved,and the difference between the Chao1 indices of the S3 and L3 groups was statistically significant(t=-2.428,P=0.022).There were not statistically differences in the alpha diversity indices between the L and S groups(P>0.05).4.The non-parametric MANOVA(Adonis)results indicated that there were significant differences in gut microbiota among the six groups(F=3.425,P=0.001),Anosim analysis showed a statistically significant difference between any two groups in group L,P<0.05.Comparing with group S0,there had statistically significant differences of S3 group and S6 group(P<0.05),but there were no statistically significant between S3 and S6 groups,P>0.05.NMDS analysis showed the value of stress=0.17(<0.2),and the results of Adonis analysis and NMDS analysis suggested that GH could regulate the composition and structure of intestinal flora of short stature children to some extent.PCoA graph and Anosim analysis showed that there were differences in species composition and community similarity between L and S groups under GH intervention,and the changes in species composition and inter-community variability were more pronounced in L groups than those in S groups.5.Species difference analysis showed a significant increase in the abundance of Enterococcaceae with long-term use of PEG-rhGH(at 6 months).Short-term use of rhGH(at 3 months)resulted in a significant decrease in the abundance of Verrucomicrobiota.6.Functional prediction analysis showed that the gut microbiota of each group were mainly involved in carbohydrate metabolism and amino acid metabolism,among other pathways.7.Correlation analysis showed that GV was negatively correlated with the Actinobacteriota,Proteobacteria,and Faecalibacterium(r=-0.565,-0.500,-0.515,P<0.05)in S3 group;GV was negatively correlated with the Prevotella(r=-0.617,P<0.05)in S6 group;GV in L6 group was negatively correlated with the Proteobacteria(r=-0.702,P<0.05).SDS for height in L6 group was positively correlated with Bacteroides and Bacteroidota/Firmicutes(B/F)ratio(r=0.583,0.794,P<0.05)and negatively correlated with Bifidobacterium(r=0.638,P<0.05);Bifidobacterium in S3 group was positively correlated with SDS for height(r=0.556,P<0.05).IGF-1 in S6 group was positively correlated with the Bacteroidota,Bacteroidota/Firmicutes(B/F)ratio,Bacteroides,and Prevotella(r=0.706,0.632,0.64,0.685,P<0.05),and negatively correlated with the Firmicutes(r=-0.560,P<0.05),and IGF-1 in L6 group was positively correlated with the Bacteroidota,the B/F ratio,Bacteroidota(r=0.637,0.736,0.577,P<0.05),and Firmicutes negatively correlated(r=-0.581,P<0.05).group L3 IGF-1 positively correlated with Bifidobacterium(r=0.562,P<0.05).Conclusions1.Growth hormone improves the growth of children with short stature,achieves growthpromoting effects,and improves IGF-1 levels.At 6 months of treatment with growth hormone,the growth-promoting effect of PEG-rhGH was more pronounced than that of rhGH.2.With the extension of growth hormone using time,although the species of dominant flora did not change,the richness,diversity and uniformity of gut microbiota are improved.PEG-rhGH is more consistent and stable than rhGH in improving the richness,diversity and uniformity of the gut microbiota.In the short term(at 3 months),PEG-rhGH shows a statistically significant advantage in improving gut microbiota richness.PEG-rhGH can significantly improve the species composition and community similarity of gut microbiota compared with rhGH.3.The continuous use of GH is able to increase the abundance of some beneficial bacteria,such as Bacteroides,and the ratio of Bacteroidota/Firmicutes on one hand,and to decrease the abundance of some harmful bacteria on the other hand.Long-term use of PEG-rhGH(at 6 months)results in a significant increase in the abundance of Enterococcaceae.Short-term use of rhGH(at 3 months)resulted in a significant decrease in the abundance of Verrucomicrobiota.4.After the use of growth hormone,some groups of beneficial flora are positively correlated with height SDS,and IGF-1 and some harmful flora are negatively correlated with GV,and the gut microbiota of each group may achieve synergistic growth-promoting effects with recombinant human growth hormone through carbohydrate metabolism and amino acid metabolism. |
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