| The theory of "Traditional Chinese medicine of heart and small intestine connection" has an important position in Chinese medicine.It reflects the relationship and communication between the heart and the small intestine in the physiological and pathological functions.At present,cardiovascular disease has become the biggest killer of human sub-health and death,and the research on cardiovascular disease is urgently needed.Atherosclerosis(AS),as one of the cardiovascular diseases,has the characteristics of high universality among the affected population and can be used as the basis of other cardiovascular diseases.With the deepening and development of microbiology research and the changes in Chinese people's dietary habits,the composition of gut microbiota and the small metabolic molecules produced by the decomposition of food have been paid more and more attention by the academic community.Among them,Trimethylamine-N-Oxide(TMAO),as a metabolite of the gut microbiota of a high-fat diet,always affects vascular health.Therefore,how to deal with the changes in the gut microbiota brought about by diet and the impact of the metabolites produced on cardiovascular health has become one of the current researches focuses of the theory of "Traditional Chinese medicine of heart and small intestine connection".Objective:Explore the TCM theory of "The heart and the small intestine connection " and the biological basis for the treatment of AS.Through the Huangdi Neijing's Alisma Orientalis Beverage(AOB)which has the effect of resolving phlegm,AS intervention is carried out while using modern biological technology to decode the"phlegm" that forms AS.Finally,it was verified that the TCM's pathological association between the gut microbiota and its metabolites and AS can be treated by AOB.Methods:First,conduct theoretical research on TCM theory of "The heart and the small intestine connection",and summarize the research direction of the relationship between the heart and the intestine and the basis of modern biology.Secondly,a theoretical summary of the literature on how the latest AS risk factor metabolite TMAO of the gut microbiota is produced and ultimately affects the "heart" through the "gut".Finally,conduct experimental research from the following four aspects:(1)The quality control analysis of AOB was carried out by high-performance liquid chromatography(HPLC)to determine whether the main components recorded in the pharmacopoeia exist in the actual decoction.(2)Determine the actual therapeutic effect of AOB on AS.ApoE-/-mice were divided into normal diet group,high-fat diet group,high-dose AOB treatment group(6.5g/kg),low-dose AOB treatment group(3.25g/kg),Antibiotic intervention group(350mg/kg),AOB-Antibiotics combined intervention group(6.5g/kg+350mg/kg),Atorvastatin treatment group(1.3mg/kg).After 8 weeks of continuous administration,the mice's body weight,body fat ratio,serum biochemical indices,the size of the pathological section of the plaque and the proportion of the lesion area were observed.(3)Clarify the regulation mechanism of AOB on TMAO,a risk factor for AS,and inflammatory factors.Under the premise of the above grouping,Western blotting was used to clarify the activity of the key enzyme FMO3 that transforms TMA into TMAO,based on the UHPLC-ESI-MS/MS platform to quantitatively analyze trimethylamine(TMA)and TMAO in serum,and use Magnetic Luminex multicytokines technology for quantitative analysis of six key inflammatory factors in serum.(4)To clarify how AOB adjusts the gut microbiota and the relationship between the changed key core gut microbiota and the risk factors of AS.After extracting and PCR amplifying the V3-V4 hypervariable part of the 16S rRNA gene of the microbiota population in the Cecal Content,sequencing was performed based on the Illumina Miseq platform,and the final sequencing results were analyzed by bioinformatics.Results:Theoretical research:through literature sorting,we have made a certain summary and generalization on the five aspects of the historical origin of TCM theory of "The heart and the small intestine connection",the meridian connection,the physiological and pathological connection,and the relationship with clinical application.At the same time,it sorted out and summarized the modern research and AOB on the TCM theory of "The heart and the small intestine connection".Secondly,from the four aspects of the production and metabolism of TMAO,the relationship between TMAO and AS,the mechanism by which TMAO promotes the formation of AS,and the use of TMAO to improve the treatment of AS,how the latest AS risk factor metabolite TMAO of the gut microbiota passes through the "gut" The final impact on the "heart" was sorted out.Experimental research:Firstly,the contents of the actual dissolution components of Alisol A and Alisol B 23-monoacetate were determined by HPLC,and the quality standards for the substance components of the aqueous decoct used in subsequent experiments were defined.In animal experiments,AOB has a relatively good therapeutic effect on various basic pathological indicators of AS regardless of high or low dose.In terms of circulating inflammatory cytokines,the use of Luminex technology can confirm that AOB effectively reduces 5 of the 6 key inflammatory cytokines that affect AS.Secondly,based on the UHPLC-ESI-MS/MS technology,the quantitative analysis of the bacterial metabolites TMAO and TMA in the serum confirmed the effective effect of AOB on TMAO,but there was little change in each group for TMA.Through western bloting analysis of the FMO3 in the liver,it was found that AOB can reduce the activity of the key enzyme FMO3 that catalyzes TMA.In addition,the Antibiotic group had no significant significance in all indicators of the effective intervention of AOB except for the reduction of body fat.However,the Antibiotic group after combined with AOB made some changes to the improvement of AS indicators.In terms of the composition of the gut microbiota,AOB improved the structure of the microbiota damaged after a high-fat diet and adjusted its richness.Combining the relative abundance of the microbiota and the LDA score can clarify that AOB can make beneficial adjustments to AS among the six bacterial genus level of Lactobacillus,Turicibacter,Eubacterium_coprostanoligenes_group,Bifidobacterium,Ileibacterium and Faecalibaculum.The highest change in relative abundance is the decrease of Lactobacillus and the increase of Bifidobacterium.After the Spearman correlation analysis with the previous indicators,the coverage of the above-mentioned two bacterial genus is also wider than the other bacterial genus.Conclusion:TCM theory of "The heart and the small intestine connection",has good practical value and reference significance in literature records,traditional theories and modern research.At the same time,experimental studies have proved that the Huangdi Neijing's AOB,which is selected in combination with the key factor "phlegm"in the formation of AS,plays a role in the adjustment of the gut microbiota,the intervention of microbiota metabolites and the ultimate protection of vascular damage Better effect.This confirms the great correlation between the heart and the small intestine in terms of the influence of the gut microbiota and its metabolites on the cardiovascular system.Therefore,our research has established a certain experimental basis for the theory of "the heart and the small intestine connection" in TCM,and has opened up horizons and ideas for future clinical exploration. |
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