Effects Of Orlistat Intervention On Clinical Endocrine Metabolism,Gut Microbiota And Fecal Short Chain Fatty Acids In Overweight And Infertile Patients With Different Ovarian Function

Part 1 Analysis of gut microbiota and short-chain fatty acids in overweight infertile patients with different ovarian functionsObjective: To understand characteristics of gut microbiota and SCFAs in overweight and infertile women with different ovarian functions,and the correlation of the different microbiota with clinical parameters and SCFAs.Methods: Recruited patients who were admitted to participate in multi-center RCT research or ask for assisted Reproductive Technology in outpatient sevice,during July 2017 to August 2018。20 patients were diagnosed PCOS with overweight,14 patients were normal ovarian function with overweight,and 12 patients were DOR with over weight.Collect clinical measurement,metabolism,and ovarian function parameters of patients,keep fecal samples,and store at-80℃ until analysis,amplify by V3-V4 region of 16 SrRNA,sequence analysis of gut microbiota using Illumina Miseq platform,and gas phase Chromatography-mass spectrometry（GC-MS）was used to detect the contents of 7 common SCFAs（acetic acid,propionic acid,butyric acid,isobutyric acid,valeric acid,isovaleric acid,caproic acid）in fecal,and to analyze different ovarian functions of overweight and infertility differences in gut microbiota and fecal SCFAs in patients,and correlations between the differential microbiota and clinical parameters and SCFAs.The IBM SPSS26.0 software was used to compare and analyze the detection indexes of different treatment groups.Results:1.Comparison of 1.Comparison of gut microbiota in overweight and infertile patients with different ovarian functions:There was no significant difference in bacterial alpha diversity index among patients in PCOS group,DOR group,and normal ovarian function group（P>0.05）;NMDS show there is no difference in bacterialβ diversity;LDA discriminant analysis showed that:Ruminococcaceae_NK4A214_group was the dominant bacteria in the overweight DOR group（LDAsocre>2）,compared between the three groups;compared with the normal ovarian function group,Lachnospiraceae_ND3007_group,Coriobacteriaceae,Collinsella,Prevotella₉ were the dominant bacteria in the PCOS group,（LDAsocre>2）,Ruminococcaceae_NK4A214_group,Coprococcus₂ are the dominant bacteria in the DOR group,（LDAsocre>2）.No significant dominant bacteria were found in the normal ovarian function group.2.Comparison of SCFAs in overweight infertile patients with different ovarian functions: the concentration of caproic acid in PCOS group was significantly lower than that in normal ovarian function group and DOR group（P<0.01）,the total concentration of SCFAs、acetic acid、 propionic acid,、isobutyric acid、butyric acid、isovaleric acid、valeric acid were no significant difference between the three groups,（P>0.05）.3.Correlation analysis of different gut microbiota with clinical parameters and SCFAs:α diversity index has no obvious correlation with clinical measurement,metabolism and hormone indexes;ACE、Chao1 index were significant positive correlation between total content of SCFAs,acetic acid,butyric acid、valeric acid,（P<0.01）.Ruminococcaceae_NK4A214,Ruminococcaceae_UCG-005,Adlercreutzia were positively correlated with the total content of SCFAs,butyric acid,valeric acid,caproic acid,（P<0.05）,the Phylum level Acidobacteria,、Dadabacteria,genus levels Bdellovibrio,Ruminococcaceae_NK4A214,Ruminococcaceae were positively correlated with caproic acid,Lachnospiraceae_ND3007_group was negatively correlated with caproic acid（P<0.05）.At the genus level,Senegalimassilia was negatively correlated with AFC,AMH,LH/FSH,（P<0.05）,positively correlated with age,FSH,FSH/LH,（P<0.001）;Ruminococcaceae_NK4A214 was negatively correlated with AFC,AMH,but FP,BMI,FSH,FSH/LH showed a significant positive correlation（P<0.05）.Coprococcus₂ was negatively correlated with AFC and AMH,and positively correlated with FP（P<0.01）.Collinsella and Coriobacteriaceae were positively correlated with AFC,waist circumference and weight.Prevotella₉ was positively correlated with AFC and FINS,（P<0.05）.Conclusion: 1.There are differences in the structure of gut microbiota in overweight infertile patients with different ovarian functions.Senegalimassilia、 Ruminococcaceae_NK4A214、 Coriobacteriaceae、 Coprococcus₂、 Collinsella,and Prevotella₉ may affect ovarian function.2.The content of caproicacid in the overweight PCOS group was lower than that in the normal ovarian function group and the DOR group,considering that caproic acid-producing bacteria in the PCOS group were different from those with normal ovarian function and DOR group.3.Collinsella,Prevotella₉,Coriobacteriaceae may be involved in the occurrence and development of chronic inflammation and polycystic ovarian changes in patients with overweight PCOS.Part 2 Effects of orlistat intervention on gut microbiota and short-chain fatty acids in patients with overweight PCOSObjective: To understand the improvement of clinical parameters,changes in gut microbiota and short-chain fatty acids of orlistat intervention and weight loss in patients with overweight PCOS.Methods:.Eighteen patients with overweight PCOS who participated in the multi-center RCT study,8 in the placebo group,and 10 in the orlistat group were collected.Before the intervention of orlistat or placebo for 1 month and 2 months of intervention The fecal specimens were amplified by the V3-V4 region of 16 Sr RNA.The Illumina Miseq platform was used to analyze the structure of the gut microbiota.The gas chromatography-mass spectrometry（GC-MS）method was used to detect the seven common SCFAs（acetic acid,propylene）in feces.Acid,butyric acid,isobutyric acid,valeric acid,isovaleric acid,caproic acid),the difference between the intestinal flora and short-chain fatty acids before and after intervention in the orlistat group and the placebo group was analyzed.IBM SPSS26.0software was used to compare and analyze the differences of various test indexes of different treatment groups.Results 1.The change of clinical pramaters before and after intervention in the orlistat group: the weight,BMI,waist circumference and hip circumference of the orlistat intervention in 1 month and 2 months were lower than that before the intervention（P<0.05）;intervention 2 There was no statistical difference between the month of January and the month of intervention（P>0.05）;there was no significant change in body weight,BMI,waist circumference and hip circumference before intervention,month of intervention and month of intervention in the placebo group（P>0.05）;There was no significant difference between the lilistat intervention group and the placebo group at different time points（P>0.05）.2.Changes in gut microbiota diversity and composition structure before and after intervention of orlistat: the intestinal flora α and β diversity indexes of placebo group and orlistat group were different at different time points between the two groups and before and after intervention There was no significant change（P>0.05）.Prior to the intervention,the different bacterial groups in the orlistat group and the placebo group were mainly Lachnospira and Hungatella.After 1 month of intervention,the relative abundance of the Orlistat group Holdemania,Hungatella,Lachnospiraceae_FCS020_group,Lachnospiraceae_UCG-010,Moryella were higher than that of the placebo group,（P<0.01）,Clostridium_sensu_stricto₁was lower than the placebo group in February,（P<0.01）,the relative abundance of Lachnospira and Klebsiella in Orlistat intervention was significantly reduced in January compared with before intervention（P<0.001）,Then maintained at a low level,compared with before and after intervention in the placebo group,compared with before intervention,Klebsiella decreased significantly after 1 month after intervention（P<0.05）;compared with before intervention,intestinal flora in 2 months after intervention The changes were mainly concentrated in the flora with relatively low abundance,and after 1 month intervention,Roseburia decreased significantly after 2 month intervention（P<0.05）.3.There was no significant difference in SCFAs between the placebo group and the orlistat group before and after intervention and between the two groups at each time point（P>0.05）.Conclusion: 1.Orlistat significantly improves the weight,body mass,waist circumference and hip circumference of patients with overweight PCOS.2.Orlistat intervention does not affect the diversity index of gut microbiota in the short term,but it has a certain effect on the reduction of obesity-associated bacteria Lachnospira,and at the same time can reduce the relative abundance of conditionally pathogenic bacteria Klebsiella.3.Orlistat intervention will not affect the content of intestinal short-chain fatty acids in the short term.