| Metabolic syndrome(MS)is a group of complex clinical symptoms caused by metabolic disorder,with a prevalence rate of 25.9%in China,which has become a serious public health challenge.Traditional Chinese medicine physique studies found that phlegm-dampness constitution is the most important constitution type to induce MS,and the risk of MS occurrence is more than 10 times that of non phlegm-dampness constitution.Therefore,it is of great significance to study the characteristics and formation mechanism of MS in phlegm-dampness constitution for reducing the occurrence of MS and ensuring people’s health.However,the disease of phlegm-dampness constitution MS is complex and difficult to be detected and paid attention to at an early stage.The exploration of simple and relatively sensitive detection methods for the diagnosis of phlegm-dampness constitution MS is beneficial to the early diagnosis and prevention of this disease.Studies have found that brown adipose tissue(BAT)has powerful thermogenic and endocrine functions,which can consume energy,generate heat,regulate glucose and lipid metabolism,so as to resist obesity,reduce the occurrence and development of type 2 diabetes and MS.The thermogenic effect of BAT can be activated by cold stimulation.A large number of studies showed that infrared thermography(IRT)technology was applied to collect infrared heat maps of BAT regions before and after cold stimulation,which could objectively reflect the activity of BAT.IRT is very sensitive to the body surface temperature.It can reflect the vitality and metabolic function of human body by collecting infrared radiation from the body surface.It is sensitive,fast,without contact and without trauma,and is suitable for the related research of phlegm-dampness MS.Traditional Chinese medicine believes that both MS and phlegm-dampness constitution are caused by the weak function of the spleen and stomach,the abnormal transport,and the internal abundance of phlegm and dampness.Phlegm and dampness accumulate in the body for a long time,leading to qi stagnation and blood stasis,thus producing a series of clinical symptoms.Therefore,this study,based on the basic theory of traditional Chinese medicine and modern pharmacological research,found three traditional Chinese medicine prescriptions that are effective in regulating the blood pressure,blood lipids and blood glucose levels of the body.They are the prescription for regulating qi and resolving phlegm-Zhizhu granules,the prescription for activating blood circulation and resolving stasis-Fufang Danshen tablets and the prescription for warming yang and removing dampness-Yiyi Fuzi powder.They conform to the therapeutic principle of phlegm-dampness constitution MS,removing dampness and phlegm,regulating qi and removing blood stasis,and can be used to treat phlegm-dampness constitution MS.The purpose of this study is to explore the characteristics of body surface thermal state of people with phlegm-dampness constitution MS by IRT technology,so as to provide a visual,objective and reliable basis for early detection and treatment of phlegm-dampness constitution MS.The network pharmacology method is used to predict the mechanism of three prescriptions in the intervention of phlegm-dampness constitution MS,providing ideas for future research.Finally,the rat model of phlegm-dampness constitution MS was constructed to explore whether traditional Chinese medicine can improve the body surface thermal state of phlegm-dampness constitution MS by increasing the expression of BAT,and to find new targets for the treatment of phlegm-dampness constitution MS with traditional Chinese medicine.Part Ⅰ Clinical trial Exploring the characteristics and formation mechanism of thermal state on the body surface of people with phlegm-dampness constitution MS based on IRT technologyObjective:To explore the characteristics of body surface thermal state and the relationship between the formation of thermal state and BAT in phlegm-dampness MS population.Method:According to the inclusion and exclusion criteria of phlegm-dampness MS,138 subjects were included and divided into three groups:healthy control group(31 cases),non phlegm-dampness constitution MS group(54 cases)and phlegm-dampness constitution MS group(53 cases).Each group was divided into young,middle-aged and old age for study.Clinical questionnaire,phlegm-dampness constitution quality subscale and basic data of subjects were collected.The relevant indicators of phlegm-dampness constitution MS diagnostic criteria were detected.The levels of BAT related metabolic indicators LP,ADP,and FGF-21 were detected.The IRT technology was used to collect the infrared thermograms of the subjects’ trunk,and 14 regions of interest(ROI)were set.Conducted the cold stimulation test on the subjects,and collected the infrared thermograms of supraclavicular region(SCR)of the subjects before and after the cold stimulation,and two ROIs were set.The professional marking tools was used to measure and analyze the average temperature of ROI body surface.Results:There were no differences in gender and age among the three groups(P>0.05),and the baseline was even.Analyzing the infrared thermograms of the trunk showed that,compared with the healthy control group,the average temperature of trunk ROI in the two MS groups decreased,compared with the non phlegm-dampness constitution MS group,the average temperature of DF and YY in the phlegm-dampness constitution MS group decreased(P<0.05 or P<0.01).Compared with the healthy control group of the same age,the average temperature of DF,YY and ZY in the young MS group was significantly decreased,the average temperature of all trunk ROIs in the middle-aged MS group was significantly decreased d(P<0.05 or P<0.01),and there was no difference in the elderly.Compared with the non phlegm-dampness constitution group of the same age,the average temperature of DF,YY and ZY in the middle-aged phlegm-dampness constitution MS group decreased(P<0.05),but no difference was found between the young and the old.Observing the infrared thermogram of SCR before and after cold stimulation,it was found that compared with before cold stimulation,the average temperature of SCR in the young and middle-aged healthy control group and the non phlegm-dampness constitution MS group increased significantly after cold stimulation,and the difference of temperature rise was significantly higher than that in the phlegm-dampness constitution MS group(P<0.01 or P<0.05),but there was no significant change in the phlegm-dampness constitution MS group;after cold stimulation,the highest temperature and the occurrence time of the highest temperature in the three groups of subjects at different ages were in the trend of healthy control group>non phlegm-dampness constitution MS group>phlegm-dampness constitution MS group.Compared with the other two groups,the levels of BMI,HOMA-IR,FINS,LP and FGF-21 in the phlegm-dampness constitution MS group increased,and the levels of BM、ADP decreased(P<0.01 or P<0.05).The average ROIs temperature of some trunk in middle-aged phlegm-dampness constitution MS group was negatively correlated with BAT-related metabolic index FINS,and positively correlated with BAT-related metabolic index FGF-21.Conclusion:The body surface thermal state of people with phlegm-dampness constitution MS showed a decrease in the average temperature of trunk ROIs,especially in DF and YY regions,the characteristics are more obvious in middle aged phlegm-dampness constitution MS.The average temperature of SCR response to cold stimulation in people of phlegm-dampness constitution MS was delayed and the temperature change was not significant,indicating that BAT activity decreased.The decrease of BAT activity in people with phlegm-dampness constitution MS has a certain correlation with the formation of thermal state of trunk ROIs.Part Ⅱ Network pharmacology research Explore the mechanism of Chinese medicine against MS based on the network pharmacology methodObjective:To understand the therapeutic targets and molecular mechanisms of Zhizhu granules,Fufang Danshen tablets and Yiyi Fuzi powder against MS.Methods:The effective ingredients with OB≥30%and DL≥0.18 were selected from the TCMSP database,and their names were normalized by Uniprot database.The keyword "metabolic syndrome" was used to search for MS related genes in GeneCards,OMIM,TTD and PharmGKB databases.Venny 2.1.0 was used to map the Venny diagram and obtain the drug-disease intersection genes.The D-C-T and PPI networks was built by STRING database and Cytoscape 3.9.1 software,and the Network Analyzer plug-in was used to analyze the networks,so as to obtain the important effective components of three prescriptions against MS,and the core targets of three prescriptions against MS were found using CytoHubba plug-in.The GO function and KEGG pathway of the intersection gene were enriched in DAVID database,and the histogram and bubble diagram of the enrichment results were drawn in the bioinformatics software.Results:Through TCMSP database screening,219 target genes of Zhizhu granules,202 target genes of Fufang Danshen tablets and 34 target genes of Yiyi Fuzi powder were obtained.Four disease databases was searched and 1032 MS related targets were obtained.Finally,69 drug-disease intersection targets in Zhizhu granules,65 in Fufang Danshen tablets and 5 in Yiyi Fuzi powder were obtained.The analysis of D-C-T network showed that the key components of Zhizhu Granule in MS intervention were quercetin,luteolin and naringin,the compound salvia miltiorrhiza tablets were quercetin,luteolin and tetrahydrotanshinone,and the coiyi aconite powder was stigmasterol,methetoin and citrostadienol.The analysis of PPI network showed that the core targets of Zhizhu granules in MS intervention were IL-6、VEGFA、TNF,ect.Fufang Danshen tablets were IL-6、TNF、VEGFA,ect,The Yiyi Fuzi powder were ESR1、EGFR、TP53,ect.Through GO and KEGG enrichment analysis,it was concluded that Zhizhu granules interfered with AGE-RAGE,lipid and atherosclerosis,insulin resistance,nonalcoholic fatty liver,endocrine and other pathways by participating in gene regulation,drug response and other biological processes.Fufang Danshen tablets interfered with AGE-RAGE,lipid and atherosclerosis,endocrine and other pathways by participating in angiogenesis,secretion factors and other processes.Yiyi Fuzi powder mainly interferes with dopaminergic synaptic signal pathway by participating in the process of dopamine catabolism and enzyme binding.Conclusion:Zhizhu granules,Fufang Danshen tablets and Yiyi Fuzi powder can intervene the occurrence and development of MS through multiple components,multiple targets and multiple pathways,mainly involving glycolipid metabolism,endocrine and dopamine pathways.Part Ⅲ Animal experiment Study on the mechanism of traditional Chinese medicine to improve the thermal state on the body surface of phlegm-dampness constitution MS rats by increasing the expression of BATObjective:To explore the influence of BAT on the body surface thermal state formation of phlegm-dampness constitution MS,and determine whether traditional Chinese medicine can improve the body surface thermal state of phlegm-dampness constitution MS by increasing the expression of BAT,and explore its mechanism.Methods:HFSSD feeding combined with intraperitoneal injection of STZ induced phlegm-dampness constitution MS rat model.After 12 weeks of modeling,the MS rats with phlegm-dampness constitution were extracted and were divided into control group,model group,Zhizhu granules group,Fufang Danshen tablets group,Yiyi Fuzi powder group and combination group,and were treated with traditional Chinese medicine for 4 weeks.The general state of rats was observed,weighed,blood pressure and abdominal circumference were measured.We observed the autonomic activity and the scapular and abdominal blood flow of rats.IRT technique was used to collect infrared thermograms of the scapula and abdomen of rats before and after cold stimulation,and the average body surface temperature was analyzed.The adipose tissue weight of different parts of rats were measured,the levels of blood glucose,blood lipid and liver function were determined,the morphology of scapular and perivascular adipose tissue of rats were observed,and UCP-1 expression in rat periscapular adipose tissue was observed.The levels of BAT related metabolic indexes,inflammatory factors and ET-1 in rats were determined,The expression levels of BAT fever and secretion related proteins were determined,the mRNA expression levels of BAT fever related genes were detected,and the levels of NO and NEFA in serum,ATP content and ATPase activity in liver were determined.Results:Compared with the control group,the rats in the model group were obese,unresponsive,slow in movement,and increased in weight,and significantly decreased in the total distance,central activity distance,and the number of cross-cuts in the open field test(P<0.05).The levels of FPG,CHO,TG,LDL-C,DBP and ALT were significantly increased(P<0.05 or P<0.01).The content of BAT in scapula decreased significantly,while the content of visceral and subcutaneous adipose tissue increased significantly(P<0.05 or P<0.01).The content of UCP-1 in scapular adipose tissue decreased,the protein expression levels of UCP-1 and PGC-1α were decreased,and the mRNA expression levels of UCP-1,PGC-1α,DIO2 and CIDEA were decreased significantly(P<0.05 or P<0.01).There was no significant change in scapular and abdominal temperature after cold stimulation,and the average temperature was significantly lower than that of control group(P<0.05 or P<0.01).PVAT morphology tended to WAT,serum NO content significantly decreased,ET-1 content significantly increased,and the blood flow of scapular significantly decreased(P<0.01).The levels of CRP,IL-6 and TNF-α was significantly higher(P<0.01),and the expression levels of LP,FGF-21 and VEGFA were significantly decreased,while the expression levels of FINS were significantly increased(P<0.05 or P<0.01).The expression of NRG-4 and FGF-21 protein decreased significantly(P<0.05 or P<0.01).Serum NEFA content was significantly increased,while ATP content and ATPase activity were significantly decreased(P<0.01).Compared with the model group,the general state of the rats in the traditional Chinese medicine group was improved,and the weight of the rats in the traditional Chinese medicine group was significantly decreased,while t ability of autonomous activity were increased.Blood glucose,blood pressure,blood lipids and abdominal circumference were improved to varying degrees.BAT content increased,visceral and subcutaneous adipose tissue decreased(P<0.05 or P<0.01).The content of UCP-1 was significantly increased(P<0.01).The protein expression and mRNA expression levels of thermogenic genes UCP-1,PGC-1α,DIO2 and CIDEA were increased(P<0.05 or P<0.01).After cold stimulation,the average temperature of scapula and abdomen increased,and the temperature was higher than that of model group(P<0.05 or P<0.01).The morphology of PVAT was significantly browned,the content of NO was significantly increased,the content of ET-1 was significantly decreased,and the blood flow status was improved(P<0.05 or P<0.01).The levels of BAT related inflammatory and metabolic indexes were regulated to a certain extent.The expression levels of FGF-21,NRG-4 and IGF-1 protein increased in different degrees(P<0.05 or P<0.01).The content of serum NEFA was significantly decreased(P<0.01),and the content of ATP and the activity of ATPase were increased in different degrees(P<0.05 or P<0.01).Conclusion:Each group of traditional Chinese medicine can improve the expression of BAT in phlegm-dampness constitution MS rats to a certain extent.Moreover,they can improve the blood flow,inflammatory reaction and glucose and lipid metabolism disorder of rats by improving the thermogenic and endocrine capacity of BAT,so as to further improve the body surface thermal state of phlegm-dampness constitution MS rats.However,the curative effect of each prescription has its own emphasis,comprehensive comparison,the best effect of the combination group.In summary,the thermal state of the body surface of people with phlegm-dampness constitution MS has characteristic manifestations,and IRT technique provides visualization and objectification basis and method for the identification and evaluation of phlegm-dampness constitution MS;BAT can be used as a new target for interfering with phlegm-dampness constitution MS and should be further studied;All three Chinese herbal prescriptions have therapeutic effects on phlegm-dampness constitution MS,but the combined prescription has the best effect,indicating that phlegm-dampness constitution MS is a complex disease requiring comprehensive intervention. |
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