Investigation Of Dietary Nutrition Status And Analysis Of Gut Microbiota Composition In Patients With Hypertensive Disorders Of Pregnancy In Chang Zhou

Objective:1.To explore the dietary nutrition status in patients with hypertensive disorders of pregnancy（HDP）in the third trimester who were selected from Changzhou,China.Factor analysis was used to build dietary patterns,and the relationships between dietary patterns and HDP were explored.2.To explore the changes of gut microbiota composition among patients with HDP,in order to provide theoretical basis for the prevention and treatment of HDP.Methods:1.A case-control study was conducted.According to the diagnostic criteria for HDP in the“Guidelines for the diagnosis and treatment of hypertensive disorders of pregnancy（2015）”,pregnant women with HDP in the third trimester who planned to deliver in the Maternal and Child Health Hospital of Changzhou were selected as the case group,and healthy pregnant women hospitalized during the same period were selected as the control group.The surveys of basic information and food frequency questionnaire（FFQ）were conducted.The dietary survey data were calculated to obtain the nutrient intake by using NCCW 12.0.According to the recommended nutrient intake（RNI）or adequate intake（AI）of pregnant women during the third trimester in the“Chinese DRIs Handbook-2013Edition”,the participants were assessed to determine whether they were at the risk of insufficient nutrient intakes.Furthermore,factor analysis was used to build dietary patterns,and the scores of dietary patterns among pregnant women with different characteristics were compared.Logistic regression models were used to analyze the relationships between the scores of different dietary patterns and HDP.Blood samples were collected to detect blood routine and biochemical indexes,and serum vitamin levels were detected by High Performance Liquid Chromatography（HPLC）.2.Fresh fecal samples were aseptically collected.Fecal bacterial genomic DNA was extracted using a kit,and the 16S r DNA V4 region sequencing was conducted based on Illumina Hi Seq 2500.The information of annotation was obtained by comparing the table of OTU abundance with the SILVA 138 database.The alpha-diversity and relative abundance of gut microbiota were estimated using the online analysis tool“Microbiomeanalyst”.The beta-diversity of gut microbiota was estimated using R 4.0.3based on Bray-Curtis distance,principal coordinate analysis was used for visualization,and permutational MANOVA was used for comparison among groups.LEf Se analysis was performed by online analysis tool based on linear discriminant analysis（LDA）.The default value of LDA was 2,and LDA score distribution histogram and cladogram were used for visualization.The heat map for correlation analysis was plotted by Hem I 1.0.Results:1.A total of 170 pregnant women in the third trimester were enrolled in this study,including 72 pregnant women with HDP and 98 pregnant women in the control group.The pre-pregnancy body mass index（BMI）,systolic blood pressure,diastolic blood pressure,and proportion of primiparas in HDP patients were significantly higher than those in the control group,whereas the gestational age was significantly lower than that in the control group（P<0.05）.In regard to hematological indexes,compared with the control group,the levels of thrombocytocrit,percentage of monocytes,percentage of basophils,glutamic oxalacetic transaminase,glutamyltranspeptidase,urea,creatinine,uric acid,cystatin C and C-reactive protein in HDP pregnant women were significantly increased,whereas the levels of total bilirubin,direct bilirubin,albumin and vitamin A among HDP subjects were significantly decreased（P<0.05）.The results of FFQ survey showed that the daily intakes of internal organs,fresh fruits,dietary vitamin A and vitamin C in HDP pregnant women were significantly lower than those in the control group（P<0.05）.The dietary fat intake of pregnant women in both groups exceeded the recommended ratio of energy,whereas the carbohydrate intake was lower than the recommended ratio of energy.The intakes of dietary fiber,vitamin B₁,calcium,iron,and selenium in both groups were lower than 80%RNI or AI,suggesting the risk of insufficient intake.In addition,patients with HDP were at the risk of protein and vitamin B₂deficiency.A total of four dietary patterns were extracted by factor analysis,which were coarse cereals and dairy pattern（rich in coarse cereals,dairy,eggs,fish,green leafy vegetables,internal organs）,seafood and mushrooms pattern（rich in shrimps,crabs and shellfish,mushrooms,poultry,nuts,fish,non-green leafy vegetables）,green leafy vegetables and fresh fruits pattern（rich in green leafy vegetables,fresh fruits,shrimps,crabs and shellfish）,rice flour and soy products pattern（rich in rice flour and other soy products）.Pregnant women with higher education were tended to choose the coarse cereals and dairy pattern,pregnant women who aged≥30 years and jobless were tended to choose the green leafy vegetables and fresh fruits pattern,pregnant women who exposed to passive smoking during pregnancy were tended to choose the rice flour and soy products pattern.The results of binary logistic regression showed that after adjusting for age,gestational age,pre-pregnancy BMI,parity,education level,occupation,sleeping time,and outdoor activities,pregnant women who got higher scores of green leafy vegetables and fresh fruits pattern were associated with a reduced risk of HDP（OR:0.53;95%CI:0.30,0.96;P<0.05）,three other dietary patterns were not associated with HDP（P>0.05）.2.A total of 129 qualified fecal samples were collected in this study,including 55 samples from the case group and 74 samples from the control group.The results showed that the beta-diversity between the two groups did not differ significantly（P=0.094）.The Shannon and chao1 indexes of gut microbiota in the case group had a decreasing trend,without statistical significance（P=0.076,0.058）.At the phylum level,the relative abundance of Bacteroidota in HDP women was significantly higher than that in the control group,whereas the relative abundance of Actinobacteriota was significantly lower than that in the control group（P<0.05）.At the family level（more than 1%）,compared with the control group,the relative abundance of Bacteroidaceae in HDP subjects was significantly increased,whereas the relative abundances of Lachnospiraceae,Prevotellaceae,Bifidobacteriaceae were significantly decreased（P<0.05）.At the genus level（more than 1%）,the relative abundance of Bacteroides in HDP women was significantly higher than that in the control group,whereas the relative abundances of Blautia,Prevotella,and Bifidobacterium were significantly lower than those in the control group（P<0.05）.The result of LEf Se analysis showed that Bacteroides was riched in HDP patients,a total of 18 bacterial genus,such as Bifidobacterium,Prevotella,Blautia,Burkholderia＿Caballeronia＿Paraburkholderia,Achromobacter were riched in the control group.For pregnant women with HDP,the relative abundance of Bacteroides was positively correlated with pre-pregnancy BMI,percentage of monocytes and parity,whereas negatively correlated with diastolic blood pressure and urea levels（P<0.05）.The relative abundance of Bautia was positively correlated with thrombocytocrit level,whereas negatively correlated with parity（P<0.05）.The relative abundance of Prevotella was positively correlated with diastolic blood pressure and glutamyltranspeptidase（P<0.05）.The relative abundance of Bifidobacterium was positively correlated with thrombocytocrit and dietary vitamin C intake（P<0.05）.Conclusion:1.The intakes of internal organs,fresh fruits,dietary vitamin A and vitamin C were lower among pregnant women with HDP.2.Green leafy vegetables and fresh fruits pattern（characterized by higher intakes of green leafy vegetables,fresh fruits,shrimps,crabs and shellfish）had a protective effect for HDP.3.The composition of gut microbiota in HDP patients changed significantly.The relative abundance of Bacteroides was increased,whereas the relative abundances of Blautia,Prevotella,Bifidobacterium were decreased.