Research On The Effect And Mechanism Of Huaji Jianpi Decoction On Glycolipid Metabolism Function And Leptin Resistance In Simple Obese Mice

Objection:In order to clarify the efficacy of Huaji Jianpi Decoction in the treatment of simple obese mice,and to explore its effect and mechanism of Huaji Jianpi Decoction on glycolipid metabolism function and leptin resistance in simple obese mice.In this study,a simple animal model of obese mice was established by feeding a high-fat diet,and the relevant indicators of modeling success were analyzed.It was treated with Chinese medicine prescription Huaji Jianpi Decoction,and western medicine Orlistat was used as a positive control.The effects of Huaji Jianpi Decoction on body weight,Lee’s index,blood glucose,insulin,blood lipids,free fatty acids,morphology of liver and adipose tissue,adipokines and the expression of related genes in JAK2-STAT3 signaling pathway were analyzed.Furthermore,the efficacy of Huaji Jianpi Decoction for weight loss was evaluated,and the possible mechanism of Huaji Jianpi Decoction in treating simple obese mice was initially clarified,and the basic experimental basis for clinical simple obesity treatment was provided.Method:1.One hundred three-week old C57BL/6N male mice were randomly divided into two groups after one week of adaptive feeding: 20 in the normal group(fed with maintenance feed)and 80 in the model group(fed with high Fat feed).The mice were fed continuously for 10 weeks and the weights of the two groups of mice were weighed and recorded every week.After 10 weeks of modeling,the body length and waist circumference of each group of mice were measured and the Lee’s index was calculated;Micro-CT was used to detect the fat distribution and volume of the two groups of mice;Automatic biochemical analyzer was used to detect the liver function enzymes,blood glucose and lipid levels of the mice and GTT and ITT experiments were performed on the two groups of mice respectively;In each group,10 animals were randomly selected for dissection,and the organs were quickly weighed and stored,and then calculated for each group of organ coefficients;The paraffin section was used to observe and analyze the differences in the morphological structure of liver and adipose tissue between two groups of mice.2.After 10 weeks of modeling,the mice were divided into 6 groups: normal group,model blank group,positive control group,Huaji Jianpi Decoction high,medium and low dose groups,10 mice in each group.The mice in each group were treated by gavage at 9:00 am every day.The positive control group was treated with western medicine orlistat.The Chinese medicine treatment group was treated with high,medium and low concentrations of Huaji Jianpi Decoction.The corresponding volume of distilled water was used in the normal group and the model blank group.During the administration,except for the normal group,the mice in the other groups were still fed with high-fat diet.3.After 6 weeks of drug intervention,the weight,body length,waist circumference,and Lee’s index of the mice in each group were compared;The fat volume of the mice in each group was measured by micro-CT;GTT and ITT experiments were used to detect the glucose tolerance(IGT)and insulin resistance(IR)of the mice in each group;Automatic biochemical analyzer was used to detect the liver function enzymes,blood glucose and lipid levels of the mice in each group;ELISA kit was used to detect the expression of INS and FFA in each group of mice;The wet weight of organ tissues was weighed during the dissection of each group of mice,and then the organ coefficient was calculated;The paraffin section was used to observe and analyze the morphological structure of liver and adipose tissue of mice in each group.4.The ELISA kit was used to detect the expression levels of Lep,ADP and IL-6 in the serum of each group of mice;RT-qPCR was used to detect the relative expression levels of LepRb,SOCS-3,JAK2 and STAT3 mRNA in liver and adipose tissue.Result:1.Ten weeks after modeling,the body weight of the model group mice increased rapidly and their body size became larger,and their body weight,body length,waist circumference,and Lee’s index were all higher than those of the normal group mice(P<0.05);The fat volume,adipose tissue wet weight and coefficient were all higher than the normal group(P<0.05);Liver tissue wet weight and liver function enzymes AST and ALT levels were higher than the normal group(P<0.05);GLU,TC,HDL,LDLand VLDL blood glucose and lipid levels were higher than normal mice(P<0.05),GTT and ITT experiments found that the blood glucose level of the model group mice was higher than that of the normal group at each test time(P<0.05),and IGT and IR appeared.In addition,the liver cells of the model group mice showed steatosis,and the volume of fat cells increased significantly with inflammatory cell infiltration.2.After 6 weeks of drug intervention,compared with the model blank group,the weight,waist circumference,Lee’s index and fat volume of the mice in each administration group decreased(P<0.05),and the adipose tissue wet weight and fat coefficient of the mice also decreased.(P<0.05);The liver tissue wet weight and liver function enzymes AST and ALT levels decreased(P<0.05),blood lipids TC,HDL,LDL and VLDL levels decreased(P<0.05),and the FFA level also decreased(P<0.05);The GLU,INS levels of mice in each administration group decreased(P<0.05),and the GTT and ITT experiments show that each administration group can improve the IGT and IR phenomenon of mice to a certain extent.Among them,western medicine has a better effect of improving IGT,and the high dose of Chinese medicine has the most significant effect of improving IR.3.Compared with the normal group,the liver cells of the model blank group mice had severe steatosis,the fat cells were significantly increased in size and uneven in size,and accompanied by obvious inflammatory cell infiltration.After drug treatment,the morphology and structure of liver and adipose tissue of mice in each administration group were improved to varying degrees compared with the model blank group.The Huaji Jianpi Decoction high and middle dose groups can effectively treat severe steatosis in liver cells of mice and significantly improve the appearance of fat hypertrophy and other conditions.4.Compared with the normal group,the levels of Lep and IL-6 in the model blank group increased(P<0.05),the ADP level decreased(P<0.05).After drug treatment,compared with the model blank group,the levels of Lep and IL-6 in mice in each administration group decreased to varying degrees(P<0.05),while ADP levels increased(P<0.05).5.Compared with the normal group,the expression levels of LepRb,JAK2 and STAT3 mRNA in liver and adipose tissue of the model blank group mice decreased(P<0.05),and SOCS-3 expression levels increased(P<0.05).After drug treatment,compared with the model blank group,Huaji Jianpi Decoction high and middle dose groups can effectively increase the expression levels of LepRb,JAK2 and STAT3 in the tissues(P<0.05),and the high,medium and low dose groups can effectively reduce the expression level of SOCS-3 gene(P<0.05),while the expression level of SOCS-3 in the positive control group is not different from the model blank group(P>0.05).Conclusion:1.After 10 weeks on a high-fat diet,the body weight,body fat,and blood lipid levels of the mice in the model group increased significantly,and problems such as IGT,IR,hepatic steatosis,and increased fat cell volume appeared,all of which indicate that the diet-induced simple obese mice have increased body fat content,and appear lipid deposition and glycolipid metabolism disorders.2.After the treatment of Huaji Jianpi Decoction,Huaji Jianpi Decoction can inhibit the weight growth of mice,reduce the Lee’s index and body fat content of mice.Huaji Jianpi Decoction can effectively regulate glycolipid metabolism disorders in obese mice by reducing blood glucose and insulin levels,improving IGT and IR,and reducing blood lipids and FFA levels in mice.3.Huaji Jianpi Decoction can effectively treat the severe steatosis in liver cells of mice,reduce ectopic deposition of fat in the body,reduce the size and volume of mouse fat cells,reduce the infiltration of inflammatory cells,and achieve the effect of weight loss and lipid reduction.4.Huaji Jianpi Decoction can reduce the level of Lep in mice,increase the level of ADP,and at the same time reduce the expression level of IL-6.It can improve hyperleptinemia in mice,and alleviate chronic inflammation and IR in mice to a certain extent.It plays a role in regulating energy homeostasis and improving metabolism disorders.5.Huaji Jianpi Decoction can increase LepRb gene expression in liver and adipose tissue,increase the binding force of Lep and LepRb,and further activate the Lep-mediated JAK2-STAT3 signaling pathway,and then by up-regulating the gene expression of LepRb,JAK2 and STAT3 in liver and adipose tissue,and down-regulating the gene expression of SOCS-3,which has the effect of improving leptin resistance and play a role in weight loss and lipid reduction.