| Objective: Obesity has gradually become a widespread public health problem,and obesity chronic low-grade inflammation can lead to the occurrence of type 2 diabetes,dyslipidemia,cardiovascular disease and cancer,etc.More and more studies have shown that the intestinal microflora and intestinal barrier function are closely related to the development of obesity chronic inflammation.In this study,we observed the effect of Zang-fu massage on the inflammatory state of patients with obesity through clinical trials,and further investigation of the effect mechanism of the vibrating abdomen method in Zang-fu massage on the chronic inflammatory state of obesity through animal experiments,and explored the possible targets of Zang-fu massage from the level of serum and intestinal inflammatory factors,the expression of intestinal barrier-related proteins and mRNAs,amino acid metabolism of intestinal microflora and their correlation with inflammatory state to provide more information about Zang-fu massage in treating obesity.Methods:1 Study on the influence of Zang-fu massage on the inflammatory state of patients with obesity: The clinical study adopted a randomized control method,and the 72 patients included were randomly divided into the treatment group(Zang-fu massage combined with lifestyle interventions)and the control group(lifestyle interventions).The massage treatment was performed 5 times a week for 4 weeks,and the changes of relevant indicators in the two groups were observed before and after treatment for 4 weeks,including body composition analysis,glucose and lipid metabolism indicators,serum inflammatory factors and TCM syndrome scores.The therapeutic effect was evaluated.BW,BMI and WC were used as the main outcome indexes,and the others were used as secondary indexes to evaluate the clinical effect of Zang-fu massage on obesity.2 Study on the effects of abdominal vibration on phenotype,glucose and lipid metabolism and inflammatory status of obese mice: After adaptive feeding,10 C57BL/6J mice were randomly selected as blank group,and the other mice were fed with high-fat diet(60% calories,Research diet No.12492)for 8 weeks to establish the obese mouse model.After successful modeling,10 mice were randomly selected as model group and abdominal vibration group.The blank group was fed with ordinary feed without any treatment,and the treatment group was mildly inhibited by isoflurane and held during the intervention.The model group was fed with high-fat diet,and other intervention methods were the same as the blank group.The massage group was continued to be fed with high-fat diet,and was treated with small animal abdominal vibrator for 10 min each time,5 times a week,for a total of 4 weeks.After 4 weeks of intervention,the weight,abdominal circumference,body fat,wet weight of fat and liver of mice were compared.HE staining was performed on the fat and liver of mice,and to evaluate the weight loss effect of abdominal shaking treatment on mice.The changes of blood lipid and blood glucose in mice were detected to evaluate the effect of abdominal vibration on the level of glucose and lipid metabolism in mice.The levels of serum inflammatory factors TNF-α、IL-10、CRP and LPS were detected by ELISA to evaluate the effect of abdominal vibration on the inflammatory state of mice.3 Study on the improvement of intestinal barrier function and its mechanism in obese mice by abdominal vibration method: The modeling method and intervention method are the same as above.After 4 weeks of intervention,the contents of inflammatory factors TNF-α,IL-10、LPS in intestinal tissue of mice were detected by ELISA;The morphological changes of intestinal tissue were observed by HE staining.The protein expression of Occludin,Claudin-1 and ZO-1 in mouse intestinal tissue was observed by immunohistochemistry.The contents of TLR4 and NF-κB protein in intestinal tissues of mice were detected by Westernblot.The mRNA expression of Occludin,Claudin-1 and ZO-1were increased,and the mRNA expression of TLR4、NF-κB in mice were detected by RT-q PCR.4 Abdominal vibration improves intestinal flora in obese mice and its association with inflammation: The modeling method and intervention method were the same as above.Macrogene sequencing technology was used to detect the intestinal flora abundance,flora composition,flora diversity,gene composition and function of mice in each group,and correlation analysis was conducted between the significantly different strains of mice in each group and their body weight,abdominal circumference,visceral fat volume,inflammatory factors of blood lipid,such as TNF-α,IL-10 and LPS.5 Study on the effect of abdominal vibration on amino acid metabolism of faeces in obese mice: The modeling method and intervention method are the same as above.Target amino acids were determined in mouse faeces,different amino acids are analyzed,and the correlation between amino acids and different strains and inflammatory factors is analyzed,so as to analyze the potential mechanism of abdominal vibration to improve the chronic inflammation of obesity.Results:1 Study on the effects of Zang-fu massage on the inflammatory state of patients with obesity:(1)Body composition analysis: After treatment,BW,BMI,WC,body fat,VFA and other levels of patients in two groups were significantly decreased compared with those before treatment(P<0.05);BW decreased more in the treatment group,and BMI,WC,body fat and VFA were significantly lower than those in the control group(P<0.05).(2)Glucolipid metabolism: Before and after treatment,the glucolipid levels in both groups were improved to varying degrees,and CHO,TG,HDL-C,LDL-C,FPG,FINS and HOMA-IR in the treatment group were significantly improved(P<0.05).Comparison between groups The treatment group was better than the control group in TG reduction(P < 0.05),but there was no significant difference between CHO,HDL-C and LDL-C groups(P>0.05).The treatment group had greater improvement on blood glucose level and insulin resistance.(3)Serum inflammatory factor levels: There were significant differences in the results of TNF-α,hs-CRP,IL-10 between the two groups before and after treatment(P<0.05),indicating that the two treatments could regulate the serum inflammation levels of the body;After treatment,the treatment group was more advantageous,and there was a significant difference between the two groups(P<0.05).(4)Serum Zonulin level: Intra-group comparison before and after treatment showed that Zonulin in both groups decreased after treatment(P<0.05),indicating that both treatments could reduce serum Zonulin level in the body;After treatment,the treatment group was more advantageous,and the difference between the two groups was significant(P<0.05).(5)TCM syndrome scores: After treatment,the total score of TCM syndrome of the two groups was significantly improved(P<0.05),and the treatment group was significantly better than the control group(P<0.05).In addition,the treatment group was also significantly better than the control group in the aspects of body obesity,limb fatigue and weakness,epigastric swelling,heavy head and body weight,thin loose stool,etc.(P<0.05).It shows that the use of Zang-fu massage intervention has more advantages in improving the symptoms of traditional Chinese medicine.(6)Efficacy evaluation: the total effective rate of the treatment group was 84.85%,and the total effective rate of the control group was 61.29%,which was better in the treatment group than in the control group(P<0.05).2 Study on the effects of abdominal vibration on the phenotype,glucose and lipid metabolism and inflammatory state of obese mice: During the whole intervention process,mice in the blank group were smaller in size,normal in mental state,diet and activity,and tight and smooth hair compared with the other two groups;In the model group,the hair of mice was rough and lacked luster,obesity and decreased mobility.The status of mice in the massage group was significantly improved compared with that in the model group,and the body weight,abdominal circumference,subcutaneous and visceral fat volume,fat and liver wet weight of mice in the massage group were significantly lower than those in the model group(P<0.01).HE staining showed that the size of fat cells of obese mice were significantly reduced after the massage intervention.The area under OGTT and ITT curves(AUC)in massage group were significantly lower than those in model group(P<0.05).After abdominal vibration treatment,the blood lipid level of mice could be improved significantly,which had significant difference compared with model group(P<0.05).The serum levels of TNF-α、LPS and CRP were significantly decreased,the serum levels of IL-10 was increased(P <0.05).3 Study on the improvement of intestinal barrier function in obese mice and its mechanism of action: After the intervention of abdominal vibration,TNF-α and LPS in intestinal tissue of mice were significantly decreased,IL-10 was increased(P<0.05).HE and immunohistochemical results showed that after abdominal vibration treatment,intestinal villus morphology of obese mice was close to normal,epithelial cells were tightly connected,mucosal structure was repaired,and the expression of intestinal barrier related tight junction proteins Occludin,Claudin-1 and ZO-1 increased.RT-q PCR showed that mRNA expressions of TLR4 and NF-κB were significantly decreased,Occludin 、Claudin-1 and ZO-1were significantly higher than those in the model group after abdominal vibration treatment(P<0.05).Western Blot analysis showed that compared with the model group,the expressions of TLR4 and NF-κB protein in the abdominal vibration group were significantly decreased(P<0.05).4 Abdominal vibration improves intestinal flora of obese mice and its association with inflammatory state: metagenomic technology is used to analyze faeces samples of mice,and the distribution of gene number among sample groups is compared.The gene richness of mice in model group is lower than that in blank group and abdominal vibration massage group.NMDS analysis showed that there were differences among the three groups at phyla,class,order,family,genus and species level,indicating that the grouping was meaningful.After the intervention,at the species level,compared to the model group,Bifidobacterium animalis,Ileibacterium valens,Bifidobacterium pseudolongum,Bifidobacterium and Bifidobacterium choerinum,Bifidobacterium cuniculi,Olsenella sp.An188,Actinobacteria bacterium UC5.1-1B11,bacterium OL-1,Collinsella Sp.4＿8＿47faa,Bifidobacterium saeculare,Eubacterium dolichum CAG375,Ktedonobacter racemifer,Paenibacillus sp.OK060,Brevibacterium linens,Anoxybacillus tepidamans,Propionibacterium acidifaciens,Erwinia toletana,Roseivirga ehrenbergii,candidate division WS6 bacterium GW2011＿GWA2＿37＿6,Candidatus Uhrbacteria bacterium The levels of GW2011＿GWD2＿52＿7 and Tetragenococcus muriaticus were significantly higher(P< 0.05).Analysis of the association with inflammatory factors found that Bifidobacterium animalis,Bifidobacterium pseudolongum,Bifidobacterium choerinum,Bifidobacterium saeculare was negatively correlated with TNF-a and LPS(P<0.05).Actinobacteria bacterium UC5.1-1B11,Eubacterium dolichum CAG375,Paenibacillus sp.OK060,Anoxybacillus tepidamans and Candidatus Uhrbacteria bacterium GW2011＿GWD2＿52＿7 were negatively correlated with LPS(P<0.05).KEGG database annotation difference analysis,the path with the highest number of annotations to level1 in the database is metabolism,Comments to level2 mainly include Carbohydrate metabolism,Amino acid metabolism,etc.5 Study on the effect of abdominal vibration on the metabolism of amino acids in the faeces of obese mice: Through the analysis of the results of targeted amino acids in the faeces of mice,it was found that abdominal vibration can regulate the metabolism of tryptophan,proline,phenylalanine and branched-chain amino acids in mice.Through analysis,it was found that glycine(Gly),tryptophan(Trp),serine(Ser),phenylalanine(Phe)and threonine(Thr)were upregulated.Methionine(Met),proline(Pro),glutamic acid(Glu),histidine(His),leucine(Leu),arginine(Arg),valine(Val)and isoleucine(Ile)were downregulated.Correlation analysis with different strains found that Gly and Bifidobacterium pseudolongum,Bifidobacterium cuniculi,Actinobacteria bacterium UC5.1-1B11,Eubacterium dolichum CAG375,Brevibacterium linens,Roseivirga ehrenbergii,Tetragenococcus muriaticus and Trp were positively correlated with Bifidobacterium(P<0.05)saeculare,Anoxybacillus tepidamans,Propionibacterium acidifaciens and Tetragenococcus muriaticus were positively correlated(P<0.05).Phe was positively correlated with Roseivirga ehrenbergii and Tetragenococcus muriaticus(P<0.05).Analysis of the association with inflammatory factors found that Glycine(Gly)was negatively correlated with LPS.Tryptophan(Trp)was negatively correlated with CRP,LPS,TNF-α and positively correlated with IL-10.Arginine(Arg)was significantly positively correlated with CRP(P < 0.05).Conclusion:1 Zang-fu massage has obvious clinical efficacy in the treatment of obesity.After the intervention of massage,BW,WC,VFA,serum inflammatory factors,Zonulin,and TCM syndrome scores of patients were significantly reduced,indicating that Zang-fu massage can significantly reduce fat and weight,inhibit obesity and low-grade inflammation and reduce intestinal permeability,and also has its unique advantages in improving TCM symptoms.2 Abdominal vibration can significantly reduce the body weight and body fat of obese mice,improve the level of lipid accumulation and glucose and lipid metabolism,and regulate the levels of serum inflammatory factors TNF-α、IL-10、LPS and CRP.3 Abdominal vibration can repair the intestinal barrier function,reduce intestinal permeability,and regulate the chronic inflammatory state of obesity,which is related to inhibiting the TLR4/NF-κB signaling pathway.4 Abdominal vibration can regulate the abundance and colony composition of intestinal flora in obese mice,and a variety of bifidobacterium strains are significantly correlated with the levels of TNF-α 、 IL-10 and LPS.Improving the translocation of intestinal flora imbalance and systemic circulation of bacteria and their products may be a potential target for the treatment of obesity inflammation.5 Abdominal vibration can regulate faeces amino acid metabolism in obese mice,and amino acids interact with intestinal flora,which plays a role in regulating intestinal barrier function and chronic inflammation. |
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