The Mechanism Of Huangqisan By Regulating Lipid Metabolism To Improve Hyperlipidemia

Hyperlipidemia refers to the abnormal increase of blood lipid levels in the blood,which is a common metabolic disease.Most patients with hyperlipidemia are accompanied by "central obesity",and the heterotopic fat accumulation is serious.The disease is usually asymptomatic and is often overlooked.With the improvement of living standard,the incidence of hyperlipidemia is higher and lower.Hyperlipidemia diseases are often linked to obesity,type 2 diabetes,heart disease,dementia,myocardial infarction,stroke,sleep apnea and atherosclerosis high mortality associated with disease,is the human health problems need to solve urgently.Compared with white fat cells containing large lipid droplets and few mitochondria,the lipid droplets in brown fat cells are small,the mitochondria are high,and there is uncoupling protein 1(UCP1).UCP1 provides brown fat(BAT)with the ability to generate heat from stored fatty acids.In the past ten years,BAT has become a hot research field,and its activity is negatively correlated with body mass index and body fat mass,which plays an important role in energy consumption and lipoprotein metabolism.Studies on mice showed that BAT activation led to the decomposition of triglycerides in cells,resulting in oxidative,uncoupled respiration and heat production,and reduced the accumulation of white fat(WAT).Currently,studies have found that WAT can appear to be brown-fat,with cold or drug stimulation,from storing energy to consuming energy,or white fat Browning.AMP activated protein kinase(AMPK)plays a major role in energy balance regulation.AMPK can increase energy consumption when activated.The AMPK signaling pathway is a classic way of regulating energy homeostasis.It regulates energy balance by combining the peripheral signals of hormones and metabolites with the neural network.Current studies confirm that AMPK is involved in feeding,brown adipose tissue,and Browning of white adipose tissue in the hypothalamus and the posterior brain by regulating the sympathetic nervous system and glucose homeostasis.The mechanism by which AMPK participates in the Browning of white fat cells is not clear.Huangqisan the radix astragali,radix puerariae,mulberry press of 1:2:1,our previous studies have shown that Huangqisan have fall fat,reduce weight,and has a good role in regulating lipid metabolism disorders,but its lipid balance mechanism of action of systemic energy has not yet been elucidated.The work is as follows:Objective Diet induced hyperlipidemia rats in this experiment,the study of Huangqisan of brown fat and white fat after the treatment(subcutaneous fat,epididymal fat)middle thermal energy related gene and protein expression;As well as to the critical path of the liver and fatty acid oxidation in skeletal muscle that AMP dependent protein kinase(AMPK)/acetyl-coa carboxylase(ACC)/carnitine fatty acyl transferaseⅠ(CPT1)signaling pathways and key targets for the synthesis of cholesterol sterol regulating element binding protein(SREBP2),HMG Co A reductase(this enzyme)the effects of which discuss Huangqisan to improve the mechanism of action of hyperlipidemia,provide the basis for Huangqisan of clinical medication.In addition,the relationship between "Browning" and "AMPK" was first explored to provide ideas for subsequent research on the mechanism of white Browning.Methods1.Therapeutic effect of Huangqisan on Hyperlipidemia Rats In this experiment,the hyperlipidemia rats induced by pure high-fat diet were established.Fifty male SD rats were fed adaptively for one week,and were randomly divided into 5 groups,10 rats in each group.In the whole feeding process,the normal group was fed with normal diet,the high fat feed model group with HFD,the low dose group with Huangqi powder with 1 g·kg-1of HQS-L,the high dose group with Astragalus powder with 2 g·kg-1HQS-H,and the positive group with Lipitorn 2 mg·kg-1.During the whole feeding process,the normal group was fed with normal diet,the other group was fed with high fat diet,and the model group was fed with high fat diet for 9 weeks.The changes of body weight,glucose metabolism,glucose tolerance,liver and skeletal muscle injury,liver steatosis and brown fat were observed 13 weeks after administration of Huangqisan in hyperlipidemic rats.Effects of epididymal fat and subcutaneous fat on histopathology,so as to explore the therapeutic effect of Huangqisan on hyperlipidemia rats.2.Effects of Huangqisan on brown fat and white fat Browning of rats with hyperlipidemia.After the study of Huangqisan,the changes of mitochondria and lipid droplet were observed in the middle heat site of brown fat.Q-PCR method is used to determine the root of remembranous milk vetch to brown fat,brown fat in the subcutaneous fat and white fat marker gene UCP1,that Prdm16,CD137,TBX1,thermogenesis gene PGC1 alpha,PGC1 beta,fatty acid oxidation CPT1 a related genes,PPAR alpha,synthesis of mitochondrial genes Tfam,NRF1,NRF2 m RNA influence;The protein expression of UCP1 in brown fat and subcutaneous fat was determined by immunohistochemistry.Method using protein western blot(WB)determination of Huangqisan of brown fat and subcutaneous fat and white fat UCP1,that Prdm16,brown marks proteins in mitochondria ATP5 A synthetic protein,which discuss Huangqisan by activating brown fat and white fat brown to improve the mechanism of action of hyperlipidemia.3.Effect of Huangqisan on the Browning effect of 3T3-L1 fat cells.In vitro experiments,3t3-l1 preadipocytes were studied and CCK method was used to determine the proliferation activity of Huangqisan on 3T3-L1 adipocytes and determine the concentration of Huangqisan in the later stage.The mature adipocytes(white adipose cells)were induced by the classical method of differentiation of 3t3-l1 fat cells.The following Huangqisan was administered to the drug for 48 h and the induction process.Q-PCR method is used to determine the root of remembranous milk vetch dispersion of mature fat cells in the marker gene of brown fat UCP1,cream-colored fat marker gene CD137 and TBX1,thermogenesis gene PGC1 alpha,PGC1 beta,fatty acid oxidation CPT1 a related genes,PPAR alpha,synthesis of mitochondrial genes Tfam,NRF1,NRF2 m RNA influence;Protein immunoblot(WB)method for determining the protein expression of UCP1 mature fat cells,thus to study the effect of Huangqisan white fat brown,targets,and the molecular mechanisms of astragalus powder to improve lipid metabolism.4.Effects of Huangqisan on AMP protein kinase signaling pathway in hyperlipidemia rats.Research Huangqisan after the treatment of the AMP depends on the critical path of fatty acid oxidation protein kinase(AMPK)/acetyl-coa carboxylase(ACC)/carnitine fatty acyl transferase Ⅰ(CPT1)signaling pathways and key targets for the synthesis of cholesterol sterol regulating element binding protein(SREBP2),HMG Co A reductase(this enzyme)m RNA and protein,the influence of mechanism to study the effect of Huangqisan lipid.Results1.Compared with the normal group,the hyperlipemia rats induced by high-fat diet can significantly increase the weight,6,Lee ’s index and serum FPG,TC,TG,LDL-C,the levels of ALT,AST,and the level of serum hdl-c leel and oral glucose tolerance is significantly reduced,hyperlipidemia rat model was established successfully.Huangqisan significantly reduced the weight,abdominal circumference and Lee’s index of rats with hyperlipidemia,and also decreased the organ coefficient of liver,epididymal fat and kidney week fat.It also significantly reduced the levels of FPG,TC,TG and ldl-c in serum of hyperlipidemia rats and improved oral glucose tolerance.At the same time,Huangqisan significantly decreased the level of AST and ALT in serum.In addition,Huangqisan can improve the pathological morphology of liver,skeletal muscle,brown fat,subcutaneous fat and epididymal fat,and the oil red O staining showed that Huangqisan decreased lipid in liver cells.The results showed that huangqisan had the effect of improving hyperlipidemia and protecting liver.2.In vivo experiment,the number of mitochondria was significantly increased after Huangqisan.The dose dependence of huangqisan increased the brown marker gene UCP1 and Prdm16 of adipose tissue,and the expression of PGC1,PGC1,and m RNA of PGC1.It also significantly increased the expression of gene CPT1 a,PPAR,CD137 and TBX1 m RNA in adipose tissue.Meanwhile,Huangqisan significantly increased the expression of gene Tfam,NRF1 and NRF2 m RNA in adipose tissue.In addition,the expression of protein UCP1,Prdm16 and ATP5A was significantly increased.Results show that the root of remembranous milk vetch scattered by increasing the number of mitochondria and active to activate brown fat,and promote the white fat brown(especially the subcutaneous fat),increase the heat production energy consumption to improve lipid metabolism and treatment of hyperlipidemia.3.In vitro experiment,Huangqisan on 3t3-L1 the lowest effective concentration is 80 μg/ml,Huangqisan to the concentration of the drug dose is 160 μg/ml,not only can significantly reduce the lipid accumulation in the mature fat cells,can significantly enhance the sign of brown fat UCP1 gene and protein expression,strengthen the sample of brown fat cells.In addition,Huangqisan also significantly increased the expression of CD137,the marker gene of PGC1,PGC1,and beige fat.The expression of CPT1 a,PPAR alpha and mitochondrial biosynthesis key enzyme gene NRF1,NRF2 and Tfam were increased.The results showed that huangqisan could brownify 3t3-l1 fat cells by increasing the expression of UCP1,and increase the mitochondrial synthesis to promote the brown-like activity,while also enhancing the oxidative energy consumption of fatty acids.4.In vivo experiment,the expression of PRKAA1,PRKAA2,ACACA,ACACB and CPT1 A m RNA of hyperlipidemia gene was increased.The expression of SREBF2 and HMGCR m RNA was decreased.The protein expressions of p-ampk,p-acc and CPT1 A were significantly increased.It had no effect on AMPK and ACC protein expression.Huangqisan decreased the protein expression of HMGCR and SREBP2.The results show that Huangqisan phosphorylates AMPK,increases the oxidation of fatty acids,inhibits cholesterol synthesis,and improves hyperlipidemia induced by high-fat diet.Conclusion1.Huangqisan significantly reduce hyperlipidemia rat body weight,blood sugar,blood fat,liver enzymes,and visceral fat,improve the pathological morphology of various organs,have the effect that improve hyperlipidemia,protect the liver.2.Huangqisan significantly increased the hyperlipidemia rats UCP1 gene and protein expression of adipose tissue and by increasing the number of mitochondria and active to activate brown fat,and promote white fat brown(especially the subcutaneous fat),increase the heat production energy consumption to improve lipid metabolism and treatment of hyperlipidemia.3.Huangqisan can increase the Browning of 3t3-l1 fat cells by increasing the expression of UCP1,and increase the mitochondrial synthesis to promote the brown-like activity,while also enhancing the oxidative energy consumption of fatty acids.4.Huangqisan activated the AMPK of hyperlipidemia rats and regulated ACC/CPT1 pathway to increase the oxidation of fatty acids and reduce the accumulation of fat.Inhibiting the expression of SREBP2 and HMGCR,inhibiting the synthesis of cholesterol,and improving hyperlipidemia induced by high-fat diet.