| Objective:This study used high-fat feed to establish an obese mouse model,and explored the mechanism of Erchen decoction in regulating glycolipid metabolism through the general condition of mice,changes in liver and epididymal fat morphology,glycolipid metabolism and changes in Chemerin/CMKLR1 signaling axis,and provided experimental basis for the mechanism of Erchen decoction in treating obese mice.Methods:Evaluation of obese mouse model: The 40 C57BL/6J mice were divided into normal group and high fat group by random number table method,including normal group(n=15)and high fat group(n=25).After 8 weeks of modeling,5 animals were randomly selected in each of the two groups,and the changes in related indicators such as apparent changes and glycolipid metabolism of mice were used as the basis for successful modeling.Animal group:normal group(n=10);Then 20 mice in the high-fat group were set as model group(n=10)and Erchen decoction group(n=10).During the 9～12th week of modeling,the normal group continued to give ordinary feed,the model group and the Erchen decoction group continued to give high-fat feed,and the mice Erchen decoction(11.1g/kg/d)were given daily gavage,and the model group and the normal group were given the same volume(20ml/kg/d)of normal saline gavage.After the intervention,the changes of IPGTT and IPITT were observed by blood glucose of mice,triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL),high-density lipoprotein cholesterol(HDL),aspartate aminotransferase(AST),and alanine aminotransferase(ALT)expression levels were detected by serum biochemistry,and morphological changes of liver tissue and epididymal adipose tissue of mice were observed by HE staining.q PCR method detected the expression levels of Chemerin,CMKLR1,GPR1 and ERK1/2 in mouse liver tissues,and WB method detected the protein expression levels of Chemerin,CMKLR1 and ERK1/2 in mouse liver tissues.Results:1.Evaluation of obese mouse model at week 8 of modeling(1)General: Compared with the normal group,the body weight,abdominal circumference,body length,and Lee’s index of the high-fat group were increased(P<0.01 or P<0.001).And the weight of mice in the high-fat group increased by more than 20%compared with the normal group.(2)Morphology of mouse liver tissue and epididymal adipose tissue: the liver tissue of mice in the high-fat group appeared round vacuoles of different sizes,the liver cord was obviously disordered,and the central venous tube appeared hyperemia.After observing the fat morphology of the epididymis of mice,the volume and number of fat cells in the hyperlipid group increased,the number of fat droplets was significantly more,the fat droplets were larger,and the nucleus was oblate oval or spindle-shaped.(3)Serum biochemical related indexes: Compared with the normal group of mice,ALT,TC,TG,HDL and LDL in the high-fat group were increased(P<0.01).(4)IPGTT and IPITT experiments: Comparing the glucose tolerance of several groups of mice,it was found that compared with the normal group,the blood glucose of the model group at each time node was significantly higher than that of the normal group of mice,and it was statistically significant(P<0.01)compared with the normal group;The area under the IPGTT blood glucose curve in the model group was significantly increased(P<0.01).Compared with the insulin tolerance of the two groups,the blood glucose levels of the model group increased significantly at 0,15,30 min and 45 min after insulin injection,and the comparison between the two groups was statistically significant(P<0.05).And the IPITT blood glucose curve of mice in the model group increased significantly,and the comparison between the two groups was statistically significant(P<0.001).2.Changes in experimental indicatorsAfter all mice were fed for 12 weeks,the following indexes were detected:(1)General conditions: the body weight,abdominal circumference,body length,and Lee’s index of mice in the model group increased compared with the normal group(P<0.05);Compared with the model group,after the intervention of Erchen decoction,the body weight,abdominal circumference,body length,and Lee’s index of mice were reduced(P<0.05).(2)Adipomorphology of liver tissue and epididymis: compared with the normal group,lipid droplets can be seen in the liver cells of the model group,the liver cord is disordered,the hepatic sinus disappears,and the central venous canal is congested;Compared with the model group,most of the liver cells in the Erchen decoction group returned to normal,the liver cord was regularly arranged,and a small number of small lipid droplets still existed in the cells.Compared with the normal group,the number of fat cells in the model group was significantly reduced,and there were large fat droplets in the model group.Compared with the model group,the volume of fat cells decreased and the number of fat cells increased in the Erchen decoction group.(3)Serum biochemical related indexes: compared with the normal group,the model group mice AST,TG,TC,LDL,HDL were significantly increased(P<0.05);Compared with the model group,the levels of AST,TG,TC and LDL were reduced(P<0.05)after the intervention of Erchen decoction,and LDL was significantly increased(P<0.05)in the model group compared with the normal group.(4)IPGTT and IPITT test: Compared with the normal group of mice,the blood glucose of the model group mice increased(P<0.01)at all time nodes,and the area under the curve increased(P<0.001);Compared with the model group,blood glucose levels decreased at 0and 120 min(P<0.05)after the intervention of Erchen decoction.The IPITT test showed that compared with the normal group,the blood glucose of each time node in the model group increased,among which,at 0,45 and 60 min,the blood glucose levels of the two groups were statistically significant(P<0.01),and the area under the curve of the model group increased significantly(P<0.001).Compared with the model group,blood glucose decreased at 45 and60 minutes(P<0.05)in the Erchen decoction group.(5)Changes in the expression of Chemerin,CMKLR1,GPR1 and ERK1/2 m RNA genes in each group of mice: compared with the normal group,the expression levels of Chemerin,CMKLR1,GPR1 and ERK1/2 m RNA in the model group were significantly increased,and the comparison between the two was statistically significant(P<0.05);Compared with the model group,the expression levels of Chemerin,CMKLR1,GPR1 and ERK1/2 m RNA decreased significantly after the intervention of Erchen decoction,and the comparison between the two was statistically significant(P<0.05).(6)Changes in the expression levels of Chemerin,CMKLR1 and ERK1/2 proteins in the liver tissues of mice in each group: compared with normal,the expression of Chemerin,CMKLR1 and ERK1/2 proteins in the liver tissues of the model group was increased(P<0.01);Compared with the model group,the expression levels of Chemerin,CMKLR1 and ERK1/2 proteins were improved after the intervention of Erchen decoction(P<0.05).Conclusions:1.Erchen decoction can regulate the general condition and glucose and lipid metabolism disorders of obese mice induced by high-fat diet,and significantly improve the morphology of liver tissue and epididymal adipose tissue.2.Obesity can lead to the disorder of Chemerin and its receptors CMKLR1,Gpr1 and downstream molecule ERK1/2 expression levels,and Erchen decoction may play a role in treating obesity by regulating this signaling axis. |
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