| Objective This research established a model of high-fat diet rats with simple obesity,some fed by orlistat capsules as contrast group,to observe the effects of Wenshen Jianpi Huatan Decoction on body weight of obese rats,Lee 's index,blood lipid,serum free fatty acids,liver and adipose tissue pathological morphology,adipokinessuch as Leptin,chemerin,heat-regulating factor UCP3 and AMPK-ACC signaling pathway related proteins and gene expression,to explore the effect of Wengan Jianpi Huatan Decoction onimproving the simple action mechanism of the lipid metabolism and Leptin resistance.Method We chose 80 male SD rats of SPF grade(weighing 120 ± 20g),selected randomly 10 of them as normal groupfed with normal diet,and the rest 70 with high fat diet.8 weeks later,the average weight ofsimple obese rats weighs 20%more than those of normal group.According to the random number table,the qualified obese rats were divided into 5 groups: Obesity model group(model group),westernmedicine orlistat treatment group(western medicine group),Wengan Jianpi Huatan Decoction low dose group(low dose group),Wenshen Jianpi Huatan Decoction middle dose group(middle dose group),Wenshen Jianpi Huatan Decoction high dose group(high dose group),There were 10 rats in each group.The rats in the low dose group are3.47 g /(kg · d),the middle dose group are 6.93 g /(kg · d),the high dose group are 13.86 g /(kg · d),and the Western medicine group are 37.8 mg /(kg · d).The normal group and the model group were fed with equal volume of distilled water,and eachboth were treated with lml / 100 g for intragastric administrationfor six weeks.During the experiment,we recorded body weight and length of the rates twice every week.By the end of the fourteenth week of the experiment,we drew blood from rats in each group,and then separated abdominal aorta and serum for detection,including rats serum total cholesterol,triglyceride,high-density lipoprotein,low density lipoproteinand the level of serum free fatty acid;serum Leptin and liver chemerin levelwereanalysed by ELISA;Cut out a small rat liver tissue and abdominal adipose tissue,then observed the liver and adipose tissue pathologyical form;theexpression of OB-R,AMPK,P-AMPK,ACC,CPT1 protein in liver and the level of UCP3 protein in skeletal muscle were analysed using Western blotting.The expression of OB-R,AMPK,ACC,CPT1 m RNA in liver and the expression of UCP3 m RNA in skeletal muscle of rats were detected by real-time fluorescence quantitative RT-PCR.Result 1.Effects of Wenshen Jianpi Huatan Decoction on lipid metabolism in simple obese ratswere listed as follows:(1)After high-fat diet for 8 weeks,the body weight and Lee's index of model group were significantly increased(p<0.01),suggesting simple obesity rats building success;(2)Wenshen Jianpi Hua-tan Decoction can reduce the body weight and Lee's index in obese rats.With administration for 6 weeks,the weight of the high and middle dosegroup were significant difference from the model group(p<0.01),Lee'sindex of the high dose groupwas significant difference from the model group(p<0.01);(3)the levels of serum TG,TC,LDL-C and FFA in the high and middle dose groups were lower than those in model group(p<0.01),the content of TG in the low dose group was different from that in th model group(p< 0.05),The content of HDL-C in the high dose group was different from that in the model group(p<0.05).2.Effects of Wenshen Jianpi Huatan Decoction on adipose tissue and liver pathological morphology in simple obese rats were listed as follows:(1)The pathological changes of adipose tissue were as followes: The area of adipocytes in middle and high dose groups was lower than that in model group,and the number of fat cells in the field of units were increased;(2)pathological changes of liver tissuewere as followes: compared with the model group,high-dose group of liver cells arranged more uniform and orderly,cytoplasmic lipid droplets smaller,cell edema and cell steatosis were improved significantly,with a small amount of inflammation Cell infiltration.3.Effects of Wenshen Jianpi Huatan Decoction on the levelof Leptin and Chemerin in simpleobese rats are listed as follows:(1)The level of the serum leptin of the model group washigher thannormal group(p<0.01),and the serum leptinin high and middle dose group were significantly higher than those inmodel group(p<0.01,p<0.05);(2)the level of chemerin in serum and liver of the model group were significantly higher than those in normal group(p<0.01);Compared with model group,the level of chemerinserum and liver were increased in the high and middle dose group(p<0.01,p<0.05).4.Effects of Wenshen Jianpi Huatan Decoction on UCP3 in Skeletal Muscle Tissue of simpleobese rats were listed as follows: The UCP3 m RNA and protein expression in skeletal muscle of model group were significantly lower than those in normal group(p<0.01);The expression of UCP3 m RNA and protein in the skeletal muscle of the model group were increased(p<0.01).5.Effects of Wenshen Jianpi Huatan Decoction on AMPK-ACC-CPT1 pathway in liver of simpleobese rats were listed as follows:(1)The expression of OB-R m RNA and protein in model group were significantly lower than those in normal group(p<0.01);compared with the model group,the expression of OB-R m RNA and protein were increased in high and middle dose group(p<0.05);(2)Compared with the normal group,the protein levels of AMPK,p-AMPK and CPT1 in the liver of the model group were lower than those in the normal group(p<0.01),the expression of AMPK and CPT1 m RNA were also decreased in model group(p<0.01);compared with model group,the levels of AMPK protein in liver of the high dose group wasincreased(p<0.05),the expression of p-AMPK protein in the liver of the high and middle dose group were significantly increased(p<0.01,p<0.05),the level of CPT1 protein in the liver of the high and middle dose group were increased(p <0.05).The levels of AMPK and CPT1 m RNA were also increased(p<0.01,p<0.05);(3)the expression of ACC protein and gene in the liver of the model group were significantly higher than those inthe normal group rat(p<0.01),the level of ACC protein and gene in the liver of the high and middle dose groups were lower than those in model group(p<0.05).Conclusion The mechanism of weight loss of Wenshen Jianpi Huatan Decoction may increase the leptin sensitivity by increasing the expression of OB-R protein in the liver.It can increase the binding force of OB-R and Leptin,and activate the Leptin-mediated AMPK-ACC-CPT1 signaling pathway,by increasing the uncoupling effect to improve basic metabolic rate.It helps to promote fatty acid oxidation and decomposition,decrease FFA levels,reduce lipid deposition,improve lipid metabolism disorders and leptin resistance. |
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