| Objective:Renal stone is a disease with a high incidence,a high recurrence rate,and a heavy economic burden.Preventing the formation and recurrence of renal stone is a huge challenge facing modern urology.Although the effect of the current Endourology satisfactory surgical treatment of kidney stones,we still lack effective measures to prevent the formation and recurrence of renal stones.It has been reported to reduce the risk of kidney stones by dietary intervention,such as the Mediterranean diet high compliance to reduce risk of people suffering from renal stones.Coffee is one of the most popular drinks in the world,but it is still uncertain whether drinking coffee can reduce the risk of renal stones.Therefore,the purpose of this study was to identify the relationships between coffee consumption and renal stone risk using large data samples from public databases and multiple analysis methods.Furthermore,we will use animal models to conduct preliminary exploration of its mechanism based on and serum metabolites.Materials and Methods:This research is carried out from the following four parts.1.The first part:We extracted large sample data from the National Health and Nutrition Examination Survey（NHANES）2007-2018,taked stone or not as the independent variable and coffee as the independent variable.We used factor weighted Logistic regression analysis of the relationship between coffee intake and the risk of renal stones.2.The second part:Systematic retrieval of the current published original research literature on the correlation between coffee and kidney stone risk.Meta-analysis was used to quantify the combined effect size to evaluate the correlation between coffee and renal stone risk.3.The third part:We Used the open large-sample Genome-Wide Association Studies（GWAS）,using the two-sample Mendelian randomization（MR）method to further clarify the relationship between coffee intake and kidney The causal relationship between the risk reduction of stones..4.The forth part:We used EG to develop renal CaOx stones model rats,observed whether coffee intervention can reduce the formation of rat kidney calcium oxalate crystals;at the same time,16s rRNA amplification and sequencing are used to analyze the intestinal flora of rats Changes;Based on the Liquid Chromatography-Mass Spectrometry Technique（LC-MS）analysis of rat serum metabolites and differences in metabolic pathways,preliminary exploration of the mechanism by which coffee inhibits the formation of renal calcium oxalate crystals.Results:1.The first part:NHANES 2007-2018 participated in the investigation of 59,842 cases,in accordance with the research,excluding the standards of 26,123 cases,and the sample was incorporated in the same amount of 183,399,374 after weighting.Compared with the non-drinking coffee crowd,1-2 cups per day（OR:0.79,95%CI:0.66-0.93,P=0.006）and 2-3 cups（OR:0.68,95%CI:0.54-0.85,P=0.001）The risk of stone is significantly reduced.Subgroup analysis shows that coffee intake still has a protective effect in certain specific people,and found 1-2 cups per day in the obese population（OR:0.73,95%CI:0.60-0.89,P=0.002）And 2-3 cups（OR:0.65,95%CI:0.50-0.83,P=0.001）Crowd of patients with non-coffee in patients with decreased stones;found in diabetic populations,daily intake of coffee is greater than 2 cups and Less than 3 cups of crowds were reduced by kidney stones in crowds by kidney stones（OR:0.54,95%CI:0.34-0.87,P=0.011）.The quadrant spacing of caffeine intake,the fourth group of people suffers from kidney stones,and the first zip group is significantly reduced by 15%（OR:0.85,95%CI:0.72-0.99,P=0.037）.Compared with non-coffee drinkers,those who consumed three or more cups of decaffeinated coffee a day had a 76 percent lower risk of developing renal stones（OR:0.24,95%CI:0.07-0.89,P=0.032）.2.The second part:After systematic retrieval and screening of inclusion and exclusion criteria,this study included a total of 9 observational studies.The total number of people included in the study was 810,316,including populations from Europe,America,and Asia.There is heterogeneity among the studies,so the random effects model is used for data merging analysis.Meta-analysis showed that the risk of kidney stones in the coffee-intake group was about 14%lower than that in the non-coffee-intake group（ES=0.859,95%CI:0.800-0.924,P<0.001）.The results of the subgroup analysis showed that the coffee consuming people in the cohort study subgroup had a 12%lower risk of kidney stones than those who did not consume coffee（ES=0.886,95%CI:0.824-0.953,P<0.001）;in the gender subgroup classification,compared with men who did not consume coffee,the risk of kidney stones in men who consumed coffee was reduced by 14%（ES=0.856,95%CI:0.809-0.965,P<0.001）.Compared with women who did not consume coffee,the risk of kidney stones in women who consumed coffee was reduced by 11%（ES=0.893,95%CI:0.818-0.975,P<0.001）;The subgroup analysis of caffeine-containing coffee intake found that compared with people who did not consume coffee,the consumption of caffeine-containing coffee（ES,0.845,95%CI:0.752-0.950,P<0.001）and decaffeinated Coffee（ES,0.887,95%CI:0.846-0.929,P<0.001）people have a lower risk of kidney stones.Funnel plot and Begg’s test analysis were used to test whether the included studies had publication bias.The results showed that the funnel plot of the included studies was basically symmetrical.The Begg’s test P value=0.171.Sensitivity analysis showed that except for one of the included literature,there was no risk of bias literature that greatly affects the stability of the combined results.3.The third part:This part studies MR analysis based on the published large sample GWAS data.Coffee gene tools are derived from the British Biological Sample Library GW AS Research（375,833 Participants,and all European blood）,through the chain imbalance（set the chain unbalance parameter R2 threshold is 0.001,genetics The distance is 10,000 kB）and a multi-effect assessment screening 7 compliant tool variables SNPS;caffeine gene tools are also from GWAS research using the British Biological Sample Library（407,072 Participants）,after the above-mentioned standard screening 12 tool variables that meet the standard SNPS.The GWAS data of renal stones is derived from the 2020 Finnish Biological Bank Database（3,856 renal stones,172,757 cases of non-kidney stones）.Using the Inverse Variance Weighting,IVW)（OR:0.986,95%CI:0.975-0.998,P=0.019）and Weighted Median Estimator,WME（OR:0.987,95%CI）:0.976-0.999,P=0.036)Analysis method found that coffee and kidney stones risk had a causal relationship.Cochran’S Q did not find a significant heterogeneity（P=0.247）,MR-EGGER Intercept test P=0.581,MR-PRESSO analysis did not find significant out of group（P=0.344）,so there is no evidence to show that we selected There is a lot effect in tool variables.Similarly,IVW（OR:0.336,95%CI:0.156-0.724,P=0.005）and WME（OR:0.386,95%CI:0.152-0.980,P=0.045）Analysis method found that caffeine and kidney stones Causal relationship.Cochran’S Q test（P=0.224）did not find a significant heterogeneity.MR-Egger Intercept test（P=0.219）and MR-PRESSO analysis（P=0.282）did not find significant off-group,so there is no evidence to show the tools we selected has multiple effect.Sensitive analysis of single study rejection method showed that the overall results were basically stable.4.The forth part:Our successful use of ethylene glycol（1%V/V）feeding rats constructed a calcium oxalate crystalline animal model（EG group）.Compared with the control group,the crystallization of the kidney tissue of the EG group was obvious.The kidney tissue lesion of Coffee and EG-Coffee group is significantly lower than that of the EG group,and the deposition of rhizophilic acid calcium acid is considered to be found in the renal tubule in part,about 20%,and there are less film.It was found that the blood creatinine had a significant statistical difference in blood creatinine,P<0.01.In addition,we found coffee to improve the destruction of the intestinal flora caused by EG.Compared with the EG group,the EG-Coffee group Alpha Diverse（Shannon Index and Shannoneven Index）is significantly higher,P<0.05;We found that the BETA diversity of the group is significantly different from the PCOA analysis（P=0.005）;We used Linear Discriminant Analysis Effect Size（LEFSE）to analyze significant differences between the EG group and the EG-Coffee group（LDA>2.5,P<0.05）.The number of bacteria related to intestinal mucosal barrier damage and metabolic diseases mediated by inflammation increased in EG group,such as Corynebacterium,Ralstonia.In the EG-Coffee group,the following beneficial bacteria:Brevibacterium、norankfMuribaculaceae、UCG003、CandidatusSoleaferrea、norankfButyricicoccaceae rising,which participated in the protection of intestinal mucosal barrier function and reduced inflammatory response,short chain fatty acid production.Metabolic syndrome analysis,found that the difference between the EG-Coffee group and the EG group was obvious in serum metabolite,mainly in the following aspects:D-Glutamine and D-glutamate metabolism,tryptopter metabolis,caffeine metabolism,sphingolipid metabolism,glycine,serine and threonine metabolism（Glycine,Serine and Threonine metabolism）,Phenylalanine metabolism. |
