Preventing And Treating Effects Of Traditional Chinese Medicine Compound GST On Fatty Liver

Fatty liver is a common disease,which is seriously harmful to human health in recent years.To explore the valid drugs of curing hepatic steatosis,new compound GST of traditional Chinese medicine is composed under the direction of Chinese medicine theries.Then we are decided to screen GST and investigate its effects of preventing and treating fatty liver and underlying mechanisms.The studies were divided into five parts:Part 1:To establish the model of hyperlipidemic fatty liver,the mice were fed with high-fat diet(HF group).The serum TC,TG,HDL-C,LDL-C and hepatic TC, TG levels were detectd from two to four weeks.At the same time,liver index was observed and hepatic steatosis was analyzed by HE and SudanⅢstaining.The results showed that serum TC,TG,HDL-C,LDL-C,TC/HDL-C and hepatic TC,TG contents were elevatd significantly in HF group two weeks after experiment.In the following three-four weeks,the serum lipid,hepatic lipid and liver index kept rising. Hepatic steatosis was induced in HF group two weeks later and the degree of steatosis was aggravated with time going on.The results suggest that feeding with high -fat diet,different phases of fatty liver in mice have been developed from two weeks to four weeks,which establishes favourable animal model for further study on drugs of anti-fatty liver.Part 2:The compound GST of traditional Chinese medicine was sieved by liver index and hepatic TG level in orthogonal design,and pharmacodynamics was verified in mice with fatty liver induced by high-fat diet for two weeks.The results indicated that the confirmed compound of improving fatty liver was composed of ginseng, panax notoginseng and gastrodia elata,which just was GST.The optimized dosage was also determined by the experiments.(Chinese patent applied No. 200910038735.2). Part 3:Hepatocyte steatosis of human liver cell line L02 was established induced by 2%alcohol and 50%calf serum,and was demonstrated by oil red staining.Then orthogonal design was used to screen main components of GST.The results showed that the optimized compound of main components was made up of ginsenosides of stem and leaf,panax notoginseng saponins and gastrodin,which obviously improved hepatocyte steatosis.Meanwhile,its optimal concentration was also confirmed. (Chinese patent applied No.200910038736.7 ).Patr 4.On the basis of part 3,the underlying mechanisms of main components of GST for ameliorating hepatocyte steatosis were investigated.The level of ROS and mitochondrial membrane potential in cells were tested by flow cytometry.Reversedphase high-performance liquid chromatography(RP-HPLC) was applied to assay cellular ATP content.Expressions of PPARαand CYP2E1 mRNA in liver were examined by RT-PCR.The results demonstrated that the effects of main components of GST on hepatocyte steatosis were associated with increasing PPARαmRNA expression in liver cells,then decreasing cellular TG level,and inhibiting CYP2EI mRNA,thus suppressing ROS level and resisting lipid peroxidation,further improving mitochondrial function and raising ATP level.(Chinese patent applied No.200910038736.7).Part 5:Mice was fed with high-fat diet for twelve weeks.When the experimental model of fatty liver was confirmed by HE staining,mice were treated intragastrically with GST,main components of GST and simvastatin respectively for two weeks. Then serum TC,TG,HDL-C,LDL-C,ALT contents,hepatic TC,TG,MDA levels and SOD activity were detected.Moreover,liver index and pathology were also observed.Expressions of PPARαand CYP2E1 mRNA in liver were examined by RT-PCR.The results indicated that GST and its main components were effective in the treatment of hepatic adipose infiltration of mice,which may be related to up-regulating PPARαmRNA,then reducing serum and hepatic TG levels,and down-regulating CYP2E1 mRNA expression in the liver,thus inhibiting lipid peroxidation.We also found that simvastatin significantly decreasd serum lipid contents,but increased hepatic lipid levels and lipid peroxidation,then aggravating fatty liver(Chinese patent applied No.200910038736.7,200910038735.2).