| Objective: Chronic low-grade inflammation is one of the characteristics of obesity.This study uses Erchen Decoction to intervene in the treatment of obesity,exploring whether Erchen Decoction can reduce obesity intestinal inflammation by regulating free mt DNA,and the correlation between Erchen Decoction and the main recognition receptors TLR9,STING,and NLRP3 of free mt DNA.Methods: 42 SPF grade C57BL/6J male mice were randomly divided into two groups based on body weight,namely a blank control group of 15 mice and a model group of 27 mice.The blank control group was fed with a normal diet powered by 4% fat,while the model group was fed with a high fat diet powered by 60% fat for 8 weeks.After modeling,three mice in each group were taken to detect the levels of serum triglyceride(TG),total cholesterol(CHO),low density lipoprotein cholesterol(LDL),high density lipoprotein cholesterol(HDL)and glucose tolerance.Determine whether the establishment of a mouse obesity model is successful based on the body mass and glycolipid metabolism of mice.Subsequently,the film-forming mice were randomly divided into a model group and an Erchen Decoction group,and a blank control group was used as a control,with 12 mice in each group.The Erchen Decoction group was given Erchen Decoction intervention,while the blank control group and model group were given equal volume physiological saline intervention,once a day,for 4 weeks.After 4 weeks,the blood lipid,glucose tolerance and hemorheology of mice in each group were detected.Realtime PCR was used to detect the expression level of mt DNA in serum.Enzyme linked immunosorbent assay(ELISA)for detecting TNF in colorectal tissue-α、IL-1 β、IL-6 levels.The m RNA and protein expression levels of TLR9,STING,and NLRP3 in mouse colorectal tissue were detected by real-time fluorescence quantitative PCR and Western blot.Results:1.Evaluation results of obesity model after 8 weeks of modeling:(1)Body mass comparison: Compared with the blank control group,the body mass of the model group mice was significantly increased(P<0.001).(2)Comparison of blood lipid indicators: Compared with the blank control group,the serum lipid indicators TG,CHO,LDL,and HDL of the model group mice were significantly increased(P<0.05).(3)Comparison of glucose tolerance: Compared with the blank control group,the blood glucose values of the model group mice increased at the 0th,15 th,30th,60 th,and 120 th min,(P<0.05).The model group mice fed with a high-fat diet showed a significant increase in body mass,which was 20% higher than the average body mass of the blank control group mice.The serum TG,CHO,LDL,and blood glucose levels of the model group mice were higher than those of the blank control group mice.Based on the above results,the obesity model in this experiment was successfully established.2.Comparison of various indicators after intervention with Erchen Decoction:(1)Body mass comparison: Compared with the blank control group,the body mass of the model group mice was significantly increased(P<0.001),and was higher than the average body mass of the blank control group mice by 20%.Compared with the model group mice,the body mass of mice in the Erchen Decoction group was significantly reduced(P<0.01).(2)Comparison of blood lipid indicators: Compared with the blank control group,the levels of serum lipid indicators TG,CHO,and HDL in the model group mice increased(P<0.05).Compared with the model group,the levels of serum lipid indicators TG,CHO,and HDL in the Erchen Decoction group decreased,with a statistically significant difference in TG(P<0.01),while there was no statistically significant difference in CHO,HDL(P>0.05).(3)Comparison of glucose tolerance: Compared with the blank control group,the blood glucose level of the model group mice increased at 0,15,30,60,and 120 min(P<0.05).Compared with the model group,the fasting blood glucose levels of mice in the Erchen Decoction group decreased(P<0.05).(4)Comparison of hemorheology related indicators: Compared with the blank control group,the model group’s hemorheology related indicators,such as erythrocyte sedimentation rate,hematocrit,plasma viscosity,and whole blood viscosity,increased,and Erchen Decoction group all decreased.The erythrocyte aggregation index and erythrocyte rigidity decreased in the model group,while they increased in the Erchen Decoction group.The above results were statistical differences(P<0.05).(5)Comparison of serum mt DNA expression levels: Compared with the blank control group,the model group mice showed an increase in serum mt DNA expression levels,with no significant difference.Compared with the model group,the serum mt DNA expression level in the Erchen Decoction group decreased,and there was no significant difference.(6)Comparison of Inflammatory factor expression levels of TNF-α 、IL-1β 、IL-6:Compared with the blank control group,in the colorectal tissue of the model group mice TNF-α、IL-1β、Increased IL-6 levels,with TNF-α There was a statistical difference(P<0.05);Compared with the model group,the serum TNF-α,IL-1β and IL-6 contents of the Erchen Decoction group were decreased,with TNF-α There was a statistical difference(P<0.05).(7)Comparison of TLR9,STING,and NLRP3 m RNA expression levels: Compared with the blank control group,the model group mice showed an increase in TLR9,STING,and NLRP3 m RNA expression levels in the colorectal tissue(P<0.01).Compared with the model group,the expression levels of TLR9,STING,and NLRP3 m RNA in the colorectal tissue of mice in the Erchen Decoction group decreased significantly(P<0.05).(8)Comparison of protein expression levels of TLR9,STING,and NLRP3: Compared with the blank control group,the expression level of STING in the colorectal tissue of mice in the model group increased significantly(P<0.01).The expression levels of TLR9 and NLRP3 were upregulated,but there was no significant difference(P>0.05).Compared with the model group,the expression levels of TLR9 and STING in the colorectal tissue of mice in the Erchen Decoction group decreased significantly(P<0.05),while the expression level of NLRP3 decreased without significant difference(P>0.05).Conclusions:The main manifestation of intestinal tissue in obese mice is the inflammatory factor TNF-α As the level increases,Erchen Decoction may reduce obesity intestinal inflammation by inhibiting the expression of free mt DNA related recognition receptors TLR9,STING,and NLRP3,with STING being the most significant. |
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