Study On The Correlation Of Gallstones With Gut Microbiota And Microbial Metabolites Short-chain Fatty Acids In Mongolian And Han People

Objective This study explores the correlation of gallstones with gut microbiota and microbial metabolites short-chain fatty acids in Mongolian and Han People,in an attempt to provide some theoretical and experimental reference for the prevention and treatment of gallostones from the microbial aspect in the future.Methods (1)We collected fecal samples of 88 volunteers from three generations of pure Mongolian and Han healthy people and gallstone patients,and then sequenced their gut microbiota by 16 S r DNA sequencing.(2)Infrared spectroscopy is used to detect the composition of gallstones in Mongolian(15 cases)and Han(15 cases)patients.(3)GC-MS method is used to determine the short chain fatty acids in feces of Han healthy people and Han gallstone patients.The relationship between gut microbiota and short chain fatty acids is analyzed by spearman correlation analysis.Results (1)A total of 88 feces of Mongolian healthy people(Q1),Han healthy people(Q2),Han patients with gallstones(Q3)and Mongolian patients with gallstones(Q4)are extracted and tested for DNA.All samples meet the high-throughput sequencing standard and are successfully detected.(2)There are differences in the composition of OTUs in different groups.Regardless of ethnicity or not,the number of OTUs in gallstone group is lower than that in healthy group.When ethnic factors are not considered,there are 2082 OTUs in the healthy person group(QI=Q1+Q2)and 1807 OTUs in the gallstone group(QII=Q3+Q4).When considering ethnic factors,there are 1907 OTUs in Q1,1311 OTUs in Q4;1606 OTUs in Q2,and 1385 OTUs in Q3.(3)Analysis of α diversity of gut microbiota: When ethnic factors are not considered,the species diversity and abundance of QII are significantly lower than QI(P<0.01).When considering ethnic factors,the species diversity and abundance of Q4 are significantly lower than that of Q1(P <0.01).But there are no significantly in species diversity and abundance between Q2 and Q3.(4)Analysis of β diversity of gut microbiota:When ethnic factors are not considered,there are significant differences in community composition between QI and QII.When considering ethnic factors,there are significant differences in community composition between Q1 and Q4.But there are no significant differences in community composition between Q2 and Q3.(5)Whether ethnic factors are considered or not,there is no significant difference in gut microbiota and the healthy group and gallstone group are mainly composed of Firmicutes and Bacteroidetes at the phylum level.Whether ethnic factors are considered are not,there is a consistent trend in the gallstone group at the level of the phylum,that means the content of Bacteroidetes in the gallstone group decreased,while the content of Proteobacteria increased.(6)At the genus level,without considering ethnic factors,unidentified＿Clostridiale,Anaerostipe,Subdoligranulum and Butyricimonas are significantly decreased in QII when compared with QI(P <0.05,P <0.01).If considering ethnic factors,Bacteroides,Faecalibacterium,unidentified＿Clostridiale,Anaerostipe,Subdoligranulum,Parabacteroides and Odoribacter significantly decreased in Q4 when compared with Q1(P <0.05,P <0.01),while Blautia significantly increased(P <0.05).Compare Q2 with Q3,Blautia,Romboutsia,Dorea,Lachnospira,Butyricicoccus and Agathobacter are significantly decreased in Q3(P <0.05,P <0.01).Compare Q1 with Q2,Alistipes,Barnesiella,Turicibacter and Odoribacter are significantly increased in Q1(P<0.05),while Fusicatenibacter,Lachnoclostridium,Lachnospira and Agathobacter are significantly decreased(P <0.05,P <0.01).(7)Cholesterol stones are the main stone components in Mongolian gallstone patients and Han gallstone patients,and there is no significant difference in stone composition between the two groups(P>0.05).(8)There are no significant difference between Q2 and Q3 in acetic acid,propionic acid,butyric acid,isobutyric acid,valeric acid,isovaleric acid and caproic acid(P > 0.05),Among them,five short-chain fatty acids including propionic acid,butyric acid,isobutyric acid,valeric acid and isovaleric acid showed a downward trend in Q3,with butyric acid having the largest decline,while acetic acid and caproic acid showed an upward trend.(9)The correlation between gut microbiota and short-chain fatty acids: Whether ethnic factors are considered or not,most of the significantly different bacterial genera that are reduced in the gallstone group are significantly correlated with short-chain fatty acids.Compare Q2 with Q3,butyric acid is significantly positively correlated with the decreased genus of butyricoccus and agathobacter in Q3(r=0.344,p=0.026;r=0.363,p=0.018).Compare Q1 with Q4,butyric acid is significantly positively correlated with the decreased genus of Faecalibacterium and Subdoligranulum in Q4(r=0.388,p=0.011;r=0.508,p=0.000).Compare QI with QII,butyric acid is significantly positively correlated with the decreased genus of Subdoligranulum and Butyricimonas in QII(r=0.508,p=0.000;r=0.344,p=0.026;).Conclusion (1)Compare gallstones patients with healthy people,the gut microbiota has obvious imbalance in gallstones patients.There are obvious differences in gut microbiota between Mongolian and Han healthy people and gallstone patients.Therefore,ethnic factors should be considered in the future microbiological treatment of gallstones.(2)Among the different bacterial genera between the Mongolian gallstone group and the Han gallstone group,Blautia is shared by the two and shows the opposite trend.Blautia significantly increased in Mongolian gallstone group and significantly decreased in Han gallstone group,which may be related to the eating habits of the two ethnic groups.(3)Cholesterol stone is the main type in Mongolian and Han gallstone patients.(4)Through the measurement results of short-chain fatty acids,which are metabolites of the gut microbiota in Han healthy group and Han gallstone group,it is found that most of the short-chain fatty acids showed a decreasing trend in the Han gallstone group,and the decreasing trend of butyric acid was the most obvious.(4)Through the analysis of the correlation between gut microbiota and short chain fatty acids,it is found that no matter whether ethnic factors are considered,the gut microbiota and short-chain fatty acids are obviously related,and the gallstone group shows a consistent trend,that is,the reduction of butyric acid bacteria.