| Objective:To explore the mechanism of Modified Yinshao Powder for nonalcoholic fatty liver disease(NAFLD)in rats by observing its effect on NAFLD,including the lipid metabolism,serum liver enzymes,hepatic oxidative stress and lipid peroxidation,hepatic histopathological changes and PERK/eIF2a/ATF4 singaling pathway in endoplasmic reticulum stress.Methods:66 SD rats were adapted for I week and the rats were randomly divided into normal group(13)and high fat diet group(53).The rats in normal group were treated with basic feed,and those in the high fat diet group were fed with high fat diet.After 8 weeks,3 rats were randomly selected from the normal group and the high fat diet group,and the liver tissues were taken for HE staining after sacrifice.The pathological changes of the liver were observed under a light microscope to observe the modeling.After confirming successful model building,the remaining 10 rats in the normal group,50 rats in the high fat diet group were randomly divided into model group,Essentiale group(Polyene Phosphatidylcholine Capsules group),high dose Chinese traditional medicine group(high dose Modified Yinshao Powder intervention group),middle dose Chinese traditional medicine group(middle dose Modified Yinshao Powder intervention group)and low dose Chinese traditional medicine group(low dose Modified Yinshao Powder intervention group),10 in each group.From the 9th week on,the rats in the Essentiale group and the Chinese traditional medicine groups were individually treated with the daily intragastric administration of Polyene Phosphatidylcholine Capsules and Modified Yinshao Powder,and the normal group and model group were given equal distilled water.After the rats were sacrificed at the end of 15th week,weighed the body weight and the liver wet weight of each group;detected the serum levels of ALT,AST,TG,TC,ALP,FFA,and MDA,SOD,TG,TC in liver of the rats;calcultated the liver index and observed pathomorphological changes in the liver tissues by HE staining.Detected each rat liver tissue ATF4 and p-eIF2a/t-eIF2a protein expression by Western-blot,and the mRNA expression level of ATF4 and eIF2a by QPCR.Results:1.The liver index,serum biochemistry,liver homogenate test results showed that compared with the model group,the serum ALT,AST,TG,TC,ALP,FFA levels,the liver index and the liver levels of MDA,TG,TC in Chinese traditional medicine groups were significantly decreased(P<0.05),the liver levels of SOD in Chinese traditional medicine groups were significantly increased(P<0.05).2.HE staining light microscope observation results showed that in the model group,the hepatic cell structure was completely destroyed and blurred,moderate-to-severe macrovesicular steatosis and the vesicular steatosis were mixed,with more common hepatocyte ballooning,inflammatory cell infiltration,and partial necrosis.In the high dose and middle dose of traditional Chinese medicine,the diffuse area,macrovesicular steatosis,and inflammatory infiltrate of hepatic tissue were significantly reduced.The hepatic cell structure of the high dose group was relatively complete,and the degree of hepatocyte lipidosis was significantly reduced,which was the closest to the normal group.3.The Western-blot results showed that the expression of ATF4 and p-eIF2a/t-eIF2a protein in hepatic tissue of NAFLD rats in each group was significantly higher than that in normal group(P<0.05).The expression of ATF4 and p-eIF2?/t-eIF2? protein in hepatic tissues of rats in Essentiale group and each dose group of traditional Chinese medicine was much down-regulated compared with the model group(P<0.05).The down-regulation of ATF4 protein expression in the high dose and middle dose groups of traditional Chinese medicine was significantly better than that in the Essentiale group(P<0.05).The down-regulatation of ATF4 protein expression in low dose Chinese medicine group was better than that in the Essentiale group(P>0.05).The down-regulation of p-eIF2?/t-eIF2? protein expression in each dose group was better than Essentiale group,but the difference was not statistically significant(P>0.05).4.The QPCR results showed that the expression of ATF4 and eIF2? mRNA in hepatic tissue of NAFLD rats in each group was significantly higher than that in normal group(P<0.05).The expression of ATF4 and eIF2? mRNA in hepatic tissues of rats in Essentiale group and each dose group of traditional Chinese medicine was much down-regulated compared with the model group(P<0.05).The down-regulation of ATF4 and eIF2a mRNA expression in each dose group of traditional Chinese medicine was significantly better than that in the Essentiale group(P<0.05).Conclusions:1.Modified Yinshao Powder can improve the general state of NAFLD rats,can reduce the serum liver enzyme and blood fat,and the excessive deposition of lipids in the liver,decrease hepatic steatosis,inflammatory infiltration,necrosis,have a certain degree of protection liver and improve the disturbance of lipid metabolism role.2.Modified Yinshao Powder can improve the liver oxidative stress status and lipid peroxidation which can decrease the degree of steatosis.3.Modified Yinshao Powder can significantly decrease the protein expression of of ATF4,p-eIF2?/t-eIF2? and mRNA expression of ATF4,eIF2a in hepatic tissue,4.Modified Yinshao Powder can decrease the degree of hepatic steatosis by inducing the excessive deposition of lipids in the liver of rats,protect the liver,reduce the liver enzyme and lipids,reflecting its good preventive and therapeutic effect on NAFLD.5.Modified Yinshao Powder can improve the liver oxidative stress and lipid peroxidation,regulate lipid metabolism,decrease hepatic steatosis,inhibit or down-regulate ATF4,p-eIF2a/t-eIF2a protein expression and ATF4,eIF2?.The mRNA expression,thereby blocking the activation of the PERK/eIF2a/ATF4 signaling pathway,exerts a therapeutic effect on NAFLD,which may be one of the mechanisms for the controlling of NAFLD by Modified Yinshao Powder. |
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