| Objective:(1)To create a Tibetan Food Frequency Questionnaire(TFFQ)which is suitable for Tibetan dietary surveys,and further evaluate its reliability and validity;(2)To investigate the dietary patterns of Tibetan residents via TFFQ,and further explore the association of Tibetan dietary patterns and hypertension;(3)To investigate the characteristics of the gut microbiota of Tibetan hypertensive patients,and further explore the association of dietary patterns and gut microbiota related to Tibetan hypertension;(4)Based on Tibetan dietary habits,to create Tibetan antihypertensive dietary prescriptions,and further evaluate its antihypertensive effect;Materials and Methods:This study adopted a multi-stage,stratified,and random sampling method to enroll1708 Tibetan residents in Luhuo County,Ganzi Prefecture.(1)The validity and reliability test of TFFQ was completed on 181 Tibetan residents.The TFFQ validity test used the 3-day 24-hour dietary recall as the reference standard,and evaluated the consistency of the results of TFFQ and the 3-day 24-hour dietary recall surveys.The TFFQ reliability test evaluated the consistency of the results of the two TFFQ surveys with an interval of 4 weeks.(2)1262 Tibetan residents(male: 38.6%;average age46.18±14.75 years old)was completed the dietary survey.All subjects completed the baseline questionnaire and TFFQ survey,physical examination and biochemical index testing.The principal component analysis(PCA)was used to extract the main dietary patterns.Each subject had a dietary pattern score for each dietary pattern,and the subjects with higher pattern score preferred this dietary pattern.(3)172 subjects with qualified stool specimens and meeting inclusion and exclusion criteria were included,then high-throughput sequencing technology was used to amplify and annotate16 Sr RNA gene V3-V4 regions in fecal genes.Linear discriminant analysis Effect Size(LEf Se)method,with LDA>2 as the threshold,was used to screen the gut microbiota related with Tibetan hypertension.Negative binomial regression model or zero inflation negative binomial regression model was used to explore the relationship between dietary pattern scores and the genus associated with Tibetan hypertension.(4)Based on the Tibetan dietary habit,referring to the recommendation of DASH Dietary Approaches to Stop Hypertension(DASH),create a Tibetan antihypertensive dietary prescription.Then a 4-week,single-center,randomized,open-label,blank-controlled study was carried out.227 Tibetan prehypertensive and grade 1 hypertensive patients were included.They were divided into dietary intervention group(114 subjects)and control group(113 subjects)by random sequence method.The dietary intervention group received a 4-week antihypertensive dietary prescriptions intervention.The control group did not receive any interventions.Intention-to-treat(ITT)analysis and per-protocol(PP)analysis were used to evaluate the antihypertensive efficacy of dietary prescriptions intervention.Results:(1)The TFFQ validity test found that the average daily intake of the food obtained by TFFQ and the 3-day 24-hour dietary recall has a good correlation.The Spearman correlation coefficient was between 0.309 and 0.690,the average correlation coefficient was 0.474.(2)The TFFQ reliability test found that the average daily intake of the food obtained by TFFQ1 and TFFQ2 had a good correlation,the Spearman correlation coefficient was between 0.377 and 0.793,and the average correlation coefficient was 0.575;intraclass correlation coefficient(ICC)was between 0.353 and 0.782,and the average ICC was 0.551.(3)Using PCA,it was found that there were three main dietary patterns among Tibetan residents: " Tsampa-red meat-potato" pattern(more intake of tsampa,beef and mutton,potatoes,butter tea,wheat and wheat products and oils),“Rice-vegetable-fruit” pattern(more intake of vegetables,rice and rice products,pork,fruit,mushrooms and eggs)and “Dairy products” pattern(more intake of dairy products).(4)Tibetan residents was classified into different dietary patterns according to their dietary pattern scores.Among them,55.2% of Tibetan residents belonged to " Tsampared meat-potato" pattern.Further comparing the differences between the three main dietary patterns in the average of the daily nutrient intake,it was found that the "Tsampa-red meat-potato" pattern had higher intake of energy,total fat and cholesterol,and lower dietary fiber intake;the "Rice-vegetable-fruit" mode had higher intake of dietary fiber and potassium;"Dairy products" pattern had a higher intake of calcium.(5)According to the dietary pattern scores,the subjects were divided into quartiles.After adjusting the confounding factors,the binary logistic regression analysis found that compared with the low quantile group(Q1 group)of the " Tsampa-red meat-potato" pattern,the high quantile group(Q4 group)significantly increased the risk of prevalence of hypertension in Tibetans,with an OR value of 3.044(95%CI: 2.060-4.500,P<0.001);Compared with the low quantile group(Q1 group)of the "Ricevegetable-fruit" pattern,the high quantile group(Q4 Group)significantly reduced the risk of the prevalence of hypertension in Tibetans,with an OR value of 0.449(95%CI:0.302-0.668,P<0.001);Compared with the low quantile group(Q1 group)of the "Dairy products " pattern,the high quantile group(Q4 Group)significantly reduced the risk of the prevalence of hypertension in Tibetans,with an OR value of OR=0.579(95%CI:0.394-0.852,P=0.005).(6)After adjusting the confounding factors,multiple linear regression analysis found that the scores of the "Tsampa-red meat-potato" pattern had an independent positive correlation with systolic blood pressure(SBP)and diastolic blood pressure(DBP)(SBP : β=2.178,P<0.001;DBP: β=1.122,P=0.001);the scores of the "Rice-vegetablefruit" pattern had an independent negative correlation with SBP and DBP(SBP: β=-2.581,P<0.001);DBP: β=-1.257,P <0.001);and the scores of the "Dairy products" pattern had an only independent negative correlation with SBP(SBP: β=-1.359,P=0.009).(7)71 Tibetan hypertensive patients and 101 normal BP subjects completed the sequencing of gut microbiota.Observed＿OTU and Shannon index were used as the index of the α diversity.Compared with the normal BP group,the hypertensive group had significantly lower Observed＿OTU and Shannon index,suggesting that Tibetan hypertensive patients had lower α diversity.The weighted Unifrac distance was used to evaluate the difference in the composition of gut microbiota,that is,the β diversity,and it was found that there was no significant difference in β diversity between the two groups.(8)LEf Se,with LDA>2 as the threshold,was used to screen the gut microbiota of Tibetan hypertension.At the phylum level,two kind of bacteria were screened out: the abundance of Firmicutes in the hypertensive group increased significantly,while the abundance of Verrucomicrobia was significantly decreased.At the genus level,12 species of bacteria were screened out: the abundance of Epulopiscium,Veillonella,Haemophilus,Megasphaera,Streptococcus and SMB53 in the hypertensive group increased significantly,while the abundance of Prevotella,Coprococcus,Ruminococcus,Dehalobacterium,Akkermansia and Wal-1855 d were significantly decreased.(9)Further explore the relationships between the different dietary patterns scores and the α diversity and the genus associated with Tibetan hypertension.After adjusting the confounding factors,it was found that the scores of the " Tsampa-red meat-potato" pattern was negatively correlated with Shannon index,the abundance of Ruminococcus and Akkermansia,while positively correlated with the abundance of Streptococcus.The scores of the "Rice-vegetable-fruit" pattern was positively correlated with Shannon index,and the abundance of Ruminococcus and Coprococcus.The scores of the "Dairy products" pattern was positively correlated with Shannon index and the abundance of Akkermansia and Ruminococcus.(10)Combining the results of Tibetan dietary survey,referring to the recommendations of DASH diet,choosing easily available local ingredients,and using salt substitute,created a nutritionally balanced and antihypertensive dietary prescription that conforms to the Tibetan dietary habits.(11)The subjects had good compliance with Tibetan antihypertensive dietary intervention(85.08%).The results of ITT analysis showed that compared with the control group,the 4-week Tibetan antihypertensive dietary intervention significantly improved office BP and ambulatory BP: the changes of office SBP/DBP were-5.98/-3.13 mm Hg;The changes of 24-h SBP/DBP were-3.23/-0.87 mm Hg;the changes of daytime SBP/DBP were-3.03/-0.73 mm Hg;the changes of nighttime SBP/DBP were-3.34/-0.97 mm Hg.The results of PP analysis was consistent with the results of ITT analysis.(12)The results of ITT analysis showed that compared with the control group,the 4-week Tibetan antihypertensive dietary intervention can significantly improve metabolic indicators: the reduction of-0.81 kg was in body weight,-0.32Kg/m2 in body mass index,-0.31umol/L in fasting blood glucose,-0.15 mmol /L in cholesterol and-0.08mmol/L in low-density lipoprotein;but had no effect on the levels of triglycerides and high-density lipoprotein.(13)The results of ITT analysis showed that compared with the control group,the 4-week Tibetan antihypertensive dietary intervention significantly reduced the-27.58mmol/d of the 24-hour urinary sodium excretion rate;significantly increased the 21.83mmol/d of the 24-hour urinary potassium excretion rate.Conclusion:(1)TFFQ has good reliability and validity,and can be used as a tool for Tibetan dietary surveys.(2)Using PCA,three main dietary patterns are found among Tibetan residents,including the " Tsampa-red meat-potato" pattern,the "Rice-vegetable-fruit" pattern and the "Dairy products" pattern.The "Tsampa-red meat-potato" pattern is a risk factor of hypertension in Tibetans,while the "Rice-vegetable-fruit" and "Dairy products" patterns are protective factors of hypertension in Tibetans.It provides theoretical basis for diet intervention in Tibetan hypertension.(3)The gut microbiota of Tibetan hypertensive patients have changes in the diversity and composition,including decreased α diversity and decreased abundance of bacteria that produce short-chain fatty acids,such as Coprococcus,Ruminococcus,and Dehalobacterium and Akkermansia;and increased the abundance of pro-inflammatory bacteria,such as Streptococcus and Veillonella.(4)Tibetan dietary patterns are significantly associated with the genus related with Tibetans hypertension.The "Rice-vegetable-fruit" and "Dairy products" pattern increase the diversity and the abundance of bacteria that produce short-chain fatty acids,while "Tsampa-red meat-potato" pattern increases the abundance of proinflammatory bacteria,and accompanied by a decrease in diversity and the abundance of beneficial bacteria.It is speculated that the Tibetan dietary patterns may be related to the prevalence of hypertension in Tibetans through gut microbiota,and providing new ideas for the exploration of the pathogenesis in Tibetan hypertension.(5)Created a Tibetan antihypertensive dietary prescription that is nutritionally balanced and in line with Tibetan dietary habits,and the Tibetan antihypertensive dietary prescription has a good antihypertensive effect and can improve metabolism,It is an effective method of dietary intervention suitable for Tibetan hypertension. |
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