Regulation Of LKB1-AMPK-ACC Signal Transduction Pathway Of Dangua Prescription Based On Insulin Resistance In HepG2 Cells

Objective:The discussion has the effect of removing blood stasis and phlegm Dangua prescription containing serum to improve the mechanism of insulin resistance and lipid deposition by regulating LKB1-AMPK-ACC signal channel,to provide the experimental basis for the basic research of traditional Chinese medicine and the treatment of diabetes and insulin resistance.Method:1.The use of cell toxicity,MTT assay and oil red O staining to observe the accumulation of lipid and glucose oxidase method to detect the sugar consumption experiment,screened the best palmitic acid and glucose induced HepG2 cell concentration and time,to establish insulin resistance and lipid accumulation model.2.The experiment was divided into 6 groups:control group,model group,solute group,metformin group,5%Dangua prescription of Dangua prescription group,10%Dangua prescription of Dangua prescription group,at the same time to intervene according to grouping 24 fluid and serum of rats induced by H.Detection of residual glucose level in the supernatant with glucose oxidase method,calculation of sugar consumption and insulin sensitive index,triglyceride content of triglyceride reagent to detect the intracellular lipid droplets,the cells were observed by oil red O staining,the expression of LKB1 was detected by Western blotting,AMPK and P-AMPK alpha,ACC and P-ACC etc.protein.Results:1.Model:the consumption of the 0.25 mmol/L 4 g/L palmitic acid and sugar concentration of 24 h HepG2 intervention by MTT experiment,oil red O staining and detection of glucose,cell activity and normal group decreased significantly(P>0.05);oil red O staining showed a large number of red cells in lipid droplets was beaded arrangement,part fusing into larger fat drops;compared with the normal group,the glucose consumption decreased significantly(P<0.01),decreased insulin sensitivity(P<0.01),said the successful establishment of the model of insulin resistance.2.Glucose consumption:compared with normal group,model group,the glucose consumption was significantly reduced(P<0.01);compared with the model group,the glucose consumption of Dangua prescription group and metformin group increased significantly(P<0.01),insulin sensitivity was increased(P<0.01),of which two metformin group,5%group of Dangua prescription 10%,Dangua prescription group the glucose consumption was not statistically significant,the insulin sensitivity index of metformin group and 10%group were higher than 5%Dangua prescription group(P>0.05).3.TG content:compared with the normal group,model group,TG content increased significantly(P<0.01);compared with the model group,Dangua prescription group and metformin group TG were significantly lower(P<0.01),the metformin group,5%group,10%Dangua prescription of Dangua prescription group the glucose consumption was not statistically significant in 10%,Dangua prescription group has the lowest TG content,5%Dangua prescription group was the highest.4.Oil red O staining:compared with the normal group,model group,red O staining of intracellular lipid droplets increased significantly and dense;compared with the model group,Dangua prescription group and metformin group after oil red O staining observation of intracellular lipid droplets decreased significantly,of which 10%of Dangua prescription group decreased most obviously.No significant difference between metformin group and 5%group of Dangua prescription.5.Western blot:The normal group and solvent control group of LKB1、P-AMPKα、AMPKα、P-ACC and ACC protein expression showed no significant difference(P>0.05);Compared with normal group,the model group of LKB1、P-AMPKα、AMPKα and P-ACC protein expression decreased(P<0.01),the expression of ACC protein increased(P<0.01);Compared with the model group,the expression of LKB1,P-AMPK,P-ACC protein in the metformin group and Dan melon group increased(P<0.01),the expression of ACC protein decreased(P<0.01),AMPK expression had no significant difference(P>0.05),by which metformin group,5%of Dangua prescription group,10%of Dangua prescription group of the protein expression showed no significant difference(P>0.05).Conclusion:1.Dangua prescription could significantly improve HepG2 cell insulin resistance glucose consumption,improve insulin sensitivity.2.The TG content of Dangua prescription could decrease the insulin resistance and lipid accumulation in HepG2 cells,improve the intracellular lipid accumulation.3.Dangua prescription could improve insulin resistance and lipid degeneration may be associated with the activation of LKB1-AMPK-ACC signaling pathway.