Study On The Effect And Mechanism Of Huaji Jianpi Decoction On Weight Loss And Lipid Reduction In Female Mice With Simple Obesity

Objection:In this study,the effect of Huaji Jianpi Decoction on obese female mice was analyzed by establishing a model of obese female mice,and detecting the body weight,Lee's index,fat volume,ITT,GTT,serum lipid and other relevant indicators,as well as the morphological observation of the liver and adipose tissue of mice.The gene expression of leptin receptor(LepRb)in fat and liver tissues and its effect on AMPK-ACC-CPT1 signaling pathway were further detected to investigate the mechanism of the anti-obesity effect of Huaji Jianpi Decoction.Methods:1.Establishment of the simple obese mouse model: 120 female C57BL/6c mice aged 3 weeks were selected,and after adaptive feeding for one week,100 mice were randomly selected and fed with high-fat diet for 10 weeks for modeling,and the remaining 20 mice were fed with ordinary diet.Mice were weighed weekly,and their body length was measured after feeding for 10 weeks.The weight of the mice in the high-fat diet group that was 20% higher than the normal diet group was selected as the model group.10 mice in the normal feed group and 10 mice in the model group were selected for ITT and GTT experiments.The fat volume and lipid level of the two groups were measured by Micro-CT and automatic biochemical analyzer.The contents of serum fatty acids,insulin,leptin,adiponectin and IL-6 were determined by Elisa.2.Take the normal group(10),44 mice in the model group were randomly divided into the model blank group(10),orlistat group(9),low-dose Chinese medicine group(8),medium-dose Chinese medicine group(8)and high-dose Chinese medicine group(9).The distilled water was used in the normal group and the model blank group.The drug group were given gavage with corresponding concentrations of drugs.After 6 weeks,the mice's body weight and Lee's index were measured.Fat volume and distribution of mice were measured by Micro-CT.The contents of TC,TG,LDL,HDL and blood glucose in serum were measured by automatic biochemical analyzer.Serum levels of FFA,INS,Leptin,ADP and IL-6 were determined by enzyme-linked immunosorbent assay(ELISA).HE staining was used to observe the histological morphology of fat and liver.3.RT-PCR was used to detect the mRNA expressions of LepRb,AMPK,CPTI and ACC in liver and adipose tissue of mice.Results:1.After 10 weeks of feeding,the mice in the model group had higher body weight and Lee's index than those in the normal group.The GTT and ITT experiments found that the model group had higher blood glucose levels at each test point than the normal group.And GTT indicated that the differences were statistically significant at 30,60,90 and 120 minutes,respectively(P < 0.05).ITT showed statistically significant differences at 0,30,60,90,and 120 minutes(P < 0.05).In terms of blood biochemical indexes,compared with the normal group,the contents of TC,TG,LDL,ALT and AST raised obviously in the model group,but the contents of HDL were significantly decreased.By ELISA,FFA,IL-6,LEP and INS were increased and ADP was decreased in the model group compared with the normal group.In addition,Micro-CT showed that the volume of subcutaneous fat and abdominal fat in normal mice was obviously lower than that in model mice.2.The normal group,the blank model group and the drug-administered group were treated by gavage.After 6 weeks,the results showed:(1)Compared with the model blank group,body weight,Lee's index and fat volume in the normal group decreased obviously.And compared with the model group,the weight,Lee's index and fat volume of the administration group all decreased decreased to varying degrees.(2)Detection of blood biochemical indicators and ELISA results indicated that,compared with the obesity model group,the levels of TC,TG,ALT,AST,LDL,FFA,INS,LEP and IL-6 in the normal group were all decreased(P < 0.05),while the levels of HDL and ADP were significantly increased(P < 0.05).And compared with the model group,the levels of TC,TG,LDL,ALT and AST in the orlistat group and the high and medium dose TCM group decreased(P < 0.05),while the levels of HDL increased(P < 0.05).The contents of TG,LDL and AST in the model group were higher than those in the low-dose group,and the result has statistical differences(P < 0.05).And compared with the model group,the FFA,INS,LEP and IL-6 levels were all decreased(P < 0.05),while ADP was significantly increased(P < 0.05).(3)The observation of the paraffin sections stained by HE showed that the number and size of lipid droplets increased significantly in the liver of the model blank group compared with the normal group.And compared with the high-fat group,the content of lipid droplets in liver of orlistat group,medium and high-dose Chinese medicine group was significantly decreased.But the effect of low dose Chinese medicine group was not obvious.And compared with the normal group,the fat cell volume in the obesity model group increased significantly,the fat cell volume in the orlistat group and the medium and high-dose Chinese medicine group decreased significantly,and the number of fat cells per unit field increased,while the effect of the low-dose Chinese medicine group was not significant.(4)The results of RT-PCR showed that LepRb expression level in liver and adipose tissue of the normal group was significantly higher than that of the model obese group(P < 0.05),and LepRb expression level in liver and adipose tissue of the model blank group was obviously lower than that of the four drug administration groups(P < 0.05).And compared with the model group,the expression levels of AMPK and CPT1 in the liver and fat of the normal group were also increased(P < 0.05).And compared with the four groups,the content of AMPK and CPT1 in the liver and fat of the model group decreased(P < 0.05).The expressions of liver and fat ACC in the normal group were lower than those in the model group(P < 0.05),and the levels of liver and fat ACC in the four groups were lower than those in the model obesity group(P < 0.05).Conclusions:1.Huaji Jianpi Decoction can inhibit the body weight,Lee's index and volume of obese female mice,reduce the levels of TC,TG,LDL and serum FFA,INS,LEP and IL-6,increase the levels of blood lipid HDL and serum ADP,and protect liver damage by reducing the contents of ALT and AST..2.The improvement of lipid metabolism in obese mice may be achieved by up-regulating the expression levels of LepRb,AMPK and CPTI in fat and liver tissues an d inhibiting the expression of ACCmRNA.