Regulatory Effect Of Hawthorn Leaves Flavonoids On Hyperlipidemia

Objective: To investigate the effect of Hawthorn Leaves Flavonoids(HLF)on blood lipids in hyperlipidemia rats and oleic acid on lipid accumulation in HepG2 cells.Methods:1.Forty-eight male SD rats were randomly divided into normal group,model group,simvastatin control group(4 mg/kg),HLF low,middle and high dose groups(100 mg / kg,200 mg/kg,400 mg/kg)with 8 rats in each group.In addition to the normal group,the other groups were given high fat diet to establish hyperlipidemia model.Drug intervention was given at the same time,the positive control group was given simvastatin at a dose of 4 mg/kg,and the HLF low,medium and high dose groups were given Hawthorn Leaves Flavonoids at a dose of 100 mg/kg,200 mg/kg and 400 mg/kg,respectively.The normal group and the model group were given the same amount of saline once a day for 8 weeks,weighing the rats weekly.After 8 weeks,the rats were killed,the content of TG,TC,HDL-C,LDL-C in plasma was detected by ELISA kit,the pathological changes of liver tissue were observed by HE staining,the lipid droplets of liver were measured by oil red O staining,and the tissue-related protein expression in liver was determined by Western blot method.2.The fat accumulation model of HepG2 cells was established by oleic acid(OA)induction in vitro for 24 h,to observe the effect of different concentrations of HLF on oleic acid-induced lipid accumulation in Hepg2 cells.The experiment was divided into normal group,model group,low,middle and high dose groups(25mg/L,50mg/L,100mg/L).After 24 h drug treatment,intracellular triglyceride(TG)content was measured by ELISA kit,lipid droplet deposition was observed by oil red O staining,and intracellular related protein expression was detected by Western blot method.Results:1.The body weight of all rats increased,the weight gain of model group was the largest,and drug groups was slower than model group,but faster than normal group.2.Compared with the normal group,the liver index,plasma levels of TG,TC and LDL-C in the model group increased significantly(P < 0.01),the lipid droplets in hepatocytes were increased,the lipid deposition was more obvious,the degree of hepatic steatosis was more serious,the expression of LKB1,p-LKB1,AMPK,p-AMPK in liver tissue decreased(p < 0.05 or p < 0.01),and the expression of ACC,p-ACC and HMGCR increased(p < 0.05 or p < 0.01)3.Compared with the model group,the plasma levels of TG,TC and LDL-c,the liver index were significantly lower,the lipid droplets inhepatocytes were significantly fewer(p < 0.05 or p < 0.01),the content of TG and TC in liver homogenate decreased to some extent,and the change of TG content in HLF middle dose group had statistical difference(p < 0.05).There was no significant difference in the expression of LKB1,AMPK,ACC protein and p-ACC,HMGCR protein expression among different groups(p < 0.05 or p < 0.01).The expression of p-LKB1,p-AMPK in HLF groups and simvastatin control group was significantly higher than that in model group(p < 0.05 or p < 0.01).4.Compared with the normal group,a large number of red lipid droplets were observed in the model group,and the intracellular TG content was significantly increased(p < 0.01),the expressions of AMPK,p-AMPK,LKB1,p-LKB1 were significantly down-regulated(p < 0.01),while the expressions of ACC,p-ACC and HMGCR were significantly up-regulated(p < 0.01),suggesting that the model of steatosis of HepG2 cells was successfully established.5.Compared with the model group,the red lipid droplets in cells of each group of HLF were significantly reduced,and the HLF could reduce the content of TG in HepG2 cells in a concentration-dependent manner,there was a significant difference between the middle and high dose groups of HLF(p < 0.05 or p < 0.01).There was no significant difference in the expression of AMPK,ACC and LKB1 between HLF groups.The expression of p-AMPK was significantly up-regulated(p < 0.05 or p < 0.01),the expression of p-ACC was significantly down-regulated(p < 0.05 or p < 0.01),the expression of p-LKB1 was significantly up-regulated in HLF meddle and high dose groups(p < 0.01),and the expression of HMGCR was significantly down-regulated in HLF high dose group(p < 0.05).Conclusions:Hawthorn Leaves Flavonoids can reduce the metabolism of blood lipids in hyperlipidemia rats induced by high fat diet,and can also affect lipid accumulation in HepG2 cells induced by oleic acid.Its mechanism may be related to inducing the protein expression of p-LKB1,p-AMPK and inhibiting the protein of p-ACC and HMGCR.