| Objective"The spleen governs the transport and transformation".The spleen can transport and transform the essence and water.High fat diet can easily hinder the spleen and stomach and damage the temper,and promote phlegm and dampness."The spleen governs the muscles." If the spleen is insufficiency,water can be transferred to the essence.And if there is no way to grow muscles,the muscle’s function will be abnormal,which accelerates the formation of phlegm and dampness.The skeletal muscle is an important metabolic site of the human body and can help to maintain the normal metabolism in the human body.Part of human glucose transport and fatty acid oxidation are carried out in skeletal muscle,which is an important place for energy metabolism and an important target organ for the treatment of obesity.Triglycerides(TG)are a manifestation of "invisible phlegm".The accumulation of triglycerides is related to weak spleen qi,muscle dystrophy,and formation of phlegm and dampness.According to TCM theory,high fat diet induced obesit obesity caused is mainly caused by spleen deficiency and phlegm.Erchen Decoction(ECD)is recorded in"Taiping Huimin Heji Bureau Prescription".The prescription is rigorous and compatible.It can regulate the qi,invigorate the spleen and dispel dampness.It is a classic prescription for strengthening the spleen and dispelling dampness.Adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK)is an important switch for regulating cell energy metabolism.In recent years,it has been found that it can inhibit acetyl Coenzyme A carboxylase(acetyl CoA carboxylase,ACC)and enhance carnitine palmitoyltransferase-Ib(carnitine palmitoyltransferase-Ib,CPT-1b)activity to reduce fatty acid synthesis and promote fatty acid oxidation(FAO).In recent reports,the AMPK/CPT-1b pathway has an important relationship with obesity prevention and treatment,and is an important research direction for obesity prevention and treatment.This study aims to explore the effect of Erchen Decoction in a high-fat diet model and its specific mechanism for preventing and treating obesity.This study revealed that body weight,blood lipid profiles,and glucose tolerance test are determined after twelves weeks treatment of Erchen decoction.The protein levels of AMPK,p-AMPK,CPT-1b in skeletal muscle are determined by western-blot.The in vitro activation of AMPK/CPT-1b pathway in C2C12 myotubes by Erchen decoction are also tested.Methods1.Study on the effect of Erchen Decoction on the body mass,blood glucose level and other indicators of high-fat diet rats and the expression level of AMPK/CPT-1b in skeletal muscle at the overall and organ levelsThirty-two male SD(sprague dawley,SD)rats,weighting 220-250g,were randomly divided into four groups:control group(CON group),high-fat diet model group(HF group),and Erchen Decoction low-dose group(ECDL group),Erchen Decoction high-dose group(ECDH group),8 rats in each group.After 7 days of adaptive feeding,the control group was fed normally,and the high-fat diet model group,the Erchen Decoction low-dose group,and the Erchen Decoction high-dose group were changed to high-fat diet.Simultaneously,gavaged with normal saline in the control group,normal saline in the high-fat diet model group,low-dose Erchen decoction(4.35 g/(kg.d))in low-dose Erchen decoction group,high-dose Erchen Decoction group was treated by high-dose Erchen Decoction(17.4 g/(kg.d)).All rats were gavaged once a day for 12 weeks.After 12 weeks,the weight of each group of rats was weighed for statistical analysis;the levels of glucose tolerance and blood lipids of each group of rats were detected;subcutaneous fat sections were sectioned and HE(hematoxylin-eosin staining,HE)stained to observe the morphological changes;western-blot(western-blot,WB)detection of AMPK phosphorylation level and CPT-1b proteins in the skeletal muscleof each group of rats changes in expression.2.Study on the effect of Erchen Decoction-containing serum on the expression of AMPK and CPT-1b in C2C12 of skeletal muscle cells at the cellular levelThe myoblasts C2C12 were induced to differentiate into skeletal muscle cells and divided into control group(CON group),5%drug-containing serum group,10%drug-containing serum group,15%drug-containing serum group,and detected AMPK,phospho-adenosine monophosphate-activated protein kinase(PAMPK),ACC,phospho-acetyl coA carboxylase(PACC)and CPT-1b protein expression levels;after C2C12 cells were induced to differentiate,they were divided into control group(CON group)and Erchen Decoction-containing serum group(ECD group),and the fluorescence expression of CPT-1b was detected.Results1.In vivo1.1 After 12 weeks of oral administration of Erchen Decoction,the weights of rats in each group were weighed.Compared with rats in the control group,the weight of the rats in the model group increased significantly,and the difference was statistically significant(P<0.05);the body mass of the low-dose Erchen Decoction group and the high-dose Erchen Decoction group was significantly lower than that of the high-fat diet model group(P<0.05).The weight of rats in the high-dose Erchen Decoction group was lower than that of the low-dose Erchen Decoction group,The difference is statistically significant(P<0.05).The area under the glucose tolerance curve of rats in the model group was significantly larger than that of the control group,and the glucose tolerance was decreased,which was statistically significant(P<0.05);the area under the glucose tolerance curve of the low-dose Erchen Decoction group and the high-dose Erchen Decoction group was higher than that of the model group.The area under the glucose tolerance curve of rats in the high-dose Erchen Decoction group was smaller than that in the low-dose Erchen Decoction group,and the difference was statistically significant(P<0.05).1.2 The triglycerides,low-density lipoproteins(LDL)and total cholesterol(CHOL)of the control group were significantly lower than those of the model group,and the difference was statistically significant(P<0.05);the triglycerides and low-density lipoproteins in the low-dose Erchen Decoction group and the high-dose Erchen Decoction group were significantly lower than the HF group,and the difference were statistically significant(P<0.05);the high-density lipoprotein(HDL)of the control group was significantly higher than the model group,and the difference was statistically significant(P<0.05);the high-density lipoprotein of rats in the high-dose Erchen Decoction group was significantly higher than that of the model group,the difference was statistically significant(P<0.05).1.3 Under 400 magnification,the voiding area of fat in the control group was smaller than that in the model group;the voiding area of fat in the model group was larger than that in the low-dose Erchen Decoction group and the high-dose Erchen Decoction group.1.4 The expression of CPT-1b protein in skeletal muscle of the model group was lower than that of the control group,and the difference was statistically significant(P<0.05).The expressions of CPT-1b protein in skeletal muscle of the low-dose and high-dose Erchen Decoction group were up-regulated compared with the model group,and the difference were statistically significant.Significance(P<0.05);the phosphorylation level of AMPK protein in the skeletal muscle of the model group was lower than that in the control group,and the difference was statistically significant(P<0.05).The phosphorylation levels of AMPK protein in the skeletal muscle of the low-dose and high-dose Erchen Decoction group were compared with that of the model group were up-regulated,and the difference were statistically significant(P<0.05).2.In vitroAfter Erchen Decoction-containing serum interfered with C2C12 skeletal muscle cells at concentrations of 5%,10%,and 15%,compared with the control group,concentrations of 5%,10%,and 15%all increased AMPK phosphorylation levels and CPT-The difference in the expression level of 1b protein was statistically significant(P<0.05)and was concentration-dependent.The up-regulation of PAMPK,PACC,and CPT-1b protein expression in 15%drug-containing serum group was greater than 10%,and 5%drug-containing serum group.After the intervention of Erchen Decoction-containing serum,the immunofluorescence detection result showed that the fluorescence expression of CPT-1b protein in Erchen Decoction-containing serum group was higher than that of the control group.Conclusions1.Erchen Decoction can reduce the body mass of high-fat diet rats and improve the glucose tolerance and blood lipid levels of high-fat diet rats;2.Erchen Decoction can up-regulate AMPK phosphorylation level and CPT-1b protein expression in skeletal muscle of high-fat diet rats in a dose-dependent manner;3.Erchen Decoction improves the effects of high-fat diet,which may be related to activating AMPK phosphorylation in skeletal muscle,inhibiting ACC,and thereby up-regulating the expression of CPT-1b protein... |
PDF Full Text Request