| ObjectiveTo explore the significance of SIRT1 expression in serum of patients with simple obesity and the mechanism of modified Erchen Decoction regulating SIRT1/NF-?B P65 signaling pathway on adipose tissue of obese C57BL/6J mice.To explore the core ideas of traditional Chinese medicine in the treatment of simple obesity and provide reference and theoretical support for the clinical prescription selection of this disease.Methods1.Data mining technologyData mining techniques were used to screen the clinical literature of Chinese medicine for simple obesity in the last 20 years.SPSS Modeler 18.0 and SPSS 24.0 statistical software were used to conduct association rule and cluster analysis for high-frequency drugs,and summarize the medication rule of Chinese prescription in treating simple obesity.2.Meta-analysisAccording to the results of data mining technology,the core treatment of simple obesity was obtained,and the clinical research literatures on the prescription treatment of simple obesity in the recent 20 years were screened,and the included standard literatures were analyzed by RevMan 5.4 software.3.Clinical partIn this part of the experiment,180 patients were selected,and serum SIRT1,BMI,waist circumference,blood lipid levels and other indicators of subjects with simple obesity and non-simple obesity were collected.Through multi-angle analysis,the significance of SIRT1 expression in patients with simple obesity was preliminarily explored.4.Animal experimentsThe mice were divided into model group,Traditional Chinese medicine group,traditional Chinese medicine+inhibitor group and agonist group,with 9 mice in each group,and 6 mice in blank group.The blank group was fed with ordinary feed.Model group was fed with high fat diet.The traditional Chinese medicine group was fed with high fat diet,supplemented erchen decoction was gavaged once a day,5 times a week.The TCM+inhibitor group was fed with high-fat diet,supplemented Erchen Decoction was gavaged once a day,and SIRT1 inhibitor was injected into tail vein 3 times a week.The agonist group was fed with high fat diet and resveratrol was administered orally 3 times a week.After 8 weeks of intervention,mice in each group were sacrificed,and tissue samples were collected and tested.Results1.After screening,216 articles and 246 traditional Chinese medicines were included.The top five frequently used drugs were Tuckahoe,Atractylodes atractylodes,tangerine peel,pinellia ternata and Hawthorn.The top five high-frequency drugs were hydrolytic drugs,qi-regulating drugs,tonifying drugs,dryness and dampness drugs and heat-clearing drugs.The proportions of the four gas frequencies from high to low were warm,cold,flat,cold and hot.The frequency ratio of five tastes was from high to low:sweet,bitter,spicy,sour,astringent and salty.The first three drugs belong to the stomach meridian,spleen meridian and lung meridian.The results of association regulation showed that atractylodes atractylodes?Tuckahoe was a common drug pair.Drug combination mainly consists of tonifying drugs and regulating qi drugs,dryness and dampness reducing phlegm drugs and regulating qi drugs.Finally,five categories were obtained by cluster analysis.2.After screening,22 RCTS were included,including 1829 patients.The results of meta-analysis showed that the clinical effective rate,BMI,body weight,waist circumference,hip circumference,abdominal circumference and waist-hip ratio of the spleen-invigorating and phlegm-huatan Zhong prescription group were superior to other treatment groups.GRADE profile3.6 showed that BMI of the spleen-invigorating and phlegm-huatanzhong prescription group was rated as high;Waist circumference,weight,chest circumference and hip circumference were rated as intermediate;Abdominal circumference and waist-to-hip ratio were rated as low.3.Clinical partThere were no significant differences in gender,smoking history,drinking history,diet,physical exercise and sleep between the control group and the case group(P>0.05).BMI,waist circumference,hip circumference,waist circumference ratio(WHR),TNF-?,SIRT1 and NF-?B were significantly different(P<0.05).The serum SIRT1 level in control group was higher than that in case group(P<0.05).The level of NF-?B in serum of control group was lower than that of control group(P<0.05).The serum TNF-? level in the control group was lower than that in the case group(P<0.05).The level of SIRT1 was negatively correlated with NF-?B,BMI,waist circumference,hip circumference,waist-hip ratio,HDL-C and TNF-?.Positive correlation with TC and LDL-C.SIRT1 was negatively correlated with simple obesity,while NF-?B and TNF-?were positively correlated with simple obesity.NF-?B,TNF-?,SIRT1 interact with NF-?B and TNF-? in simple obesity.4.Animal experimentsExperiment 1:The results showed that after 8 weeks of modeling,the weight difference between the two groups was statistically significant(P<0.01).After intervention,the body weight,fat weight and body fat index of model group were higher than those of blank group(P<0.05).Compared with model group,TCM decoction group had lower body weight,fat weight and body fat index(P<0.05);There was no significant difference between TCM+SIRT1 inhibitor group and model group(P>0.05).Compared with model group,the improvement degree of fat weight and body fat index in agonist group was higher(P<0.05).Experiment 2:The adipose tissue of the blank group was normal and the cell volume was small.Compared with the normal group,the volume of adipocytes in the model group was significantly increased,and the number of adipocytes in the model group was significantly decreased,while the number of adipocytes in the TCM group was significantly increased.The morphology,volume and number of adipocytes in TCM plus inhibitor group did not change significantly.In agonist group,the volume of adipocytes decreased and the number of adipocytes increased significantly.Compared with blank group,the levels of TC,TG and LDL-C in model group increased,while hdL-C decreased.The levels of TC,TG and LDL-C decreased and HDL-C increased(P<0.05)after treatment with Modified Erchen Decoction.Compared with model group,the serum lipid level of agonist group was also decreased(P<0.05),and the TC and TG levels of obese mice were improved in TCM plus inhibitor group(P<0.05).Experiment 3:Compared with blank group,IDO value and M1/M2 value of CD86 macrophages in model group were increased,while IDO value of CD206 macrophages was decreased(P<0.05);Compared with model group,IDO value and M1/M2 value of CD86 macrophages in TCM group were decreased,while IDO value of CD206 macrophages was increased(P<0.05).Compared with model group,there were no significant differences in IDO and M1/M2 values of CD86 macrophages and CD206 macrophages in TCM+inhibitor group(P>0.05):Compared with model group,IDO and M1/M2 values of CD86 macrophages in agonist group were decreased,while IDO and M1/M2 values of CD206 macrophages were increased(P<0.05).Experiment 4:Compared with blank group,the levels of TNF-?,IL-6 and SFRP5 in serum of model group were significantly different(P<0.05).Compared with model group,TNF-?,IL-6 and SFRP5 in TCM group were changed in different degrees(P<0.05).Compared with model group,the levels of TNF-?,SFRP5 and IL-6 in TCM plus inhibitor group had no significant changes(P>0.05).Compared with model group,TNF-?,IL-6 and SFRP5 levels in agonist group were changed(P<0.05).Compared with blank group,the expression levels of NF-?B,NF-?B P65,P-I ?Ba,TNF-?,IL-6 protein and related genes in model group were increased(P<0.05),while the expression levels of SFPR5 protein and related genes were decreased(P<0.05).Compared with model group,the expressions of NF-?B,NF-?B P65,P-I ?Ba,TNF-?,IL-6 protein and related genes in TCM group were decreased,while the expressions of SFPR5 protein and related genes were increased(P<0.05).Compared with model group,the expressions of NF-?B,NF-?B P65,P-I ?Ba,TNF-?,IL-6 protein and related genes in agonist group were decreased,while the expressions of SFPR5 protein and related genes were increased in agonist group(P<0.05).There were no significant differences in NF-?B,NF-?B P65,P-I ?Ba,TNF-?,IL-6,SFPR5 protein and related genes between TCM+inhibitor group and model group(P<0.05).Experiment 5:Compared with blank group,the expression of SIRT1 protein and corresponding gene in adipose tissue of model group was decreased,the fluorescence intensity of SIRT1 was lower,the fluorescence intensity of NF-?B P65 was increased(P<0.05),and the SIRT1/NF-?B P65 ratio was decreased(P<0.05).Compared with model group,the expression of SIRT1 protein and corresponding genes in adipose tissue of TCM group were increased,the fluorescence intensity of SIRT1 was increased,the fluorescence intensity of NF-?B P65 was decreased,and the ratio of SIRT1/NF-?B P65 was increased(P<0.05).Compared with model group,there were no statistically significant differences in SIRT1 protein and related gene expression,SIRT1 and NF-?B P65 fluorescence intensity(P>0.05),and SIRT1/NF-?B P65 ratio(P>0.05)in TCM plus inhibitor group.Compared with model group,the expression of SIRT1 protein and related genes,SIRT1 fluorescence intensity,NF-?B p65 fluorescence intensity and SIRT1/NF-?B P65 ratio in agonist group were increased(P<0.05).Conclusion1.Modern doctors treat simple obesity by invigorating the spleen and reducing phlegm as the core therapy,taking into account drugs such as promoting blood circulation,eliminating food,clearing heat and purging.2.The prescription of invigorating spleen and resolving phlegm in treating simple obesity is effective and worthy of clinical promotion.3.The serum SIRT1 level of simple obesity patients was lower than that of normal population.The levels of NF-?B and TNF-? in patients with simple obesity were higher than those in normal population.Serum SIRT1 levels were negatively correlated with the incidence of simple obesity,while NF-?B and TNF-? levels were positively correlated with the incidence of simple obesity.SIRT1 interacts with NF-?B and TNF-? on simple obesity.4.Modified Erchen Decoction can improve obesity degree and inhibit adipocyte hyperplasia in obese mice,possibly through activating SIRT1,inhibiting NF-?B p65 nuclear transport and reducing downstream inflammatory factor expression. |
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