The Molecular Mechanism Of Huayu Qutan Decoction Regulating Mitochondrial Energy Metabolism And Interfering With Liver Lipid Deposition In AS Rabbits

Purpose:Exploring the mechanism of Huayu Qutan Recipe which improves the liver lipid deposition by affecting the mitochondrial energy metabolism in AS rabbits to enrich the scientific connotation of “phlegm and blood stasis” theory in AS.Material and method:90 healthy SPF male white New Zealand rabbits were randomly divided into the normal group(15 rabbits),the Huayu Qutan Recipe high-dose group(15 rabbits),the Huayu Qutan Recipe medium-dose group(15 rabbits),the Huayu Qutan Recipe low-dose group(15rabbits)and the simvastatin group(15 rabbits)after fed 2 weeks of acclimation.Then,the normal group was continued fed with common diet,and normal saline was injected intravenously into the ear margin simultaneously.The other groups were fed with high-fat-diet,and bovine serum albumin were injected intravenously into the era margin simultaneously.All rabbits in each group were free to eat and drink water.Eight weeks later,the experimental group was randomly divided into and were given to the corresponding drug solution according to conversion coefficient of surface area between human and animal.The normal group and the model group were given to the same amount of normal saline.All groups were administered intragastrically once a day for 4 weeks.The content of TC,TG,LDL-C and HDL-C in serum were detected by the automatic biochemistry analyzer.HE and oil red O staining were uesed to observe the changes of liver tissue morphology.The activity of liver mitochondrial respiratory chain enzyme complexs was detected by colorimetric method.The content of mitochondrial respiratory chain enzyme complexes was detected by Blue-Native-Page.The content of ATP in liver was detected by Elisa.The levels of mitochondrial energy metabolism-related genes in liver were detected by PCR array.The mitochondrial energy metabolism-related proteins were detected by Western blot.Results:1.Effect of Huayu Qutan recipe on blood lipid level in AS Rabbits Compared with the normal group,the levels of TC,TG and LDL-C were significantly increased in the model group(P<0.01),while the HDL-C level was significantly decreased in the model group(P<0.01);Compared with the model group,the levels of TC,TG and LDL-C in the high,medium and low dose of Huayu Qutan Recipe and the simvastatin group were lower significantly(P<0.01),and HDL-C level was higher significantly(P<0.01).The treatment effect of the Huayu Qutan Recipe high-dose group and the simvastatin group was better than other treatment groups(P<0.01 or P<0.05).2.Effect of Huayu Qutan recipe on liver histomorphology and lipid deposition in AS Rabbits Compared with the normal group,the liver tissue cells in the model group were obviously swollen,a large number of fat vacuoles were clearly visible,and there was obvious accumulation of orange lipid droplets in the cytoplasm of the rabbits in the model group.Compared with the model group,the hepatic fat vacuole-like degeneration and lipid deposition in each group were better to some extent.While,the Huayu Qutan recipe high dose group and the simvastatin group were better than other groups.3.Effect of Huayu Quthan recipe on Energy Metabolism of liver mitochondria in AS Rabbits Results of mitochondrial respiratory chain enzyme complexs activity: Compared with the normal group,the activity of hepatic mitochondrial respiratory chain enzyme complex I,II,III,IV decreased in the model group(P<0.05),and compared with the model group,the activity of hepatic mitochondrial respiratory chain enzyme complex I,II,III,IV in the Huayu Qutan recipe high dose group and the simvastatin group were increased(P<0.05).Blue-native-page results: compared with the normal group,the content of mitochondrial respiratory chain enzyme complex I,II,III,IV,V in the model group was decreased(P<0.05),and compared with the model group,the content of hepatic mitochondrial respiratory chain enzyme complex I,II,III,IV,V in the Huayu Quphan recipe high dose group and the simvastatin group were increased(P<0.05).Elisa results: compared with the normal group,the ATP content in the liver of the model group was decreased(P<0.01),and the ATP content in the liver tissue of the Huayu Quphan recipe high dose group and the simvastatin group was increased compared with the model group(P<0.05).4.Effect of Huayu Qutan recipe on Mitochondrial Energy Metabolism related genes and protein.PCR array results: compared with the normal group,73 up-regulated genes and 6down-regulated genes were obtained in the model group.Compared with the model group,10up-regulated genes and 70 down-regulated genes were obtained in the Huayu Qutan recipe high-dose group.Of the 67 genes up-regulated in the model group,46 genes were reversed after treatment by Huayu Qutan recipe.Western Blot results: compared with the normal group,the expression of NDUFS1 ?NDUFS2?UQCRC1?UQCRC2?ATP5C1?COX4I1?COX6B1 in the model group decreased(P<0.05 or P<0.01).Compared with the model group,the expression of NDUFS1?NDUFS2?UQCRC1?UQCRC2?ATP5C1?COX4I1?COX6B1 in the Huayu Qutan recipe high-dose group was higher than that in the model group(P<0.05 or P <0.01).Conclusion:1.Huayu Qutan recipe can reduce the level of blood lipid in AS rabbits and improve liver lipid deposition.2.Huayu Qutan recipe can promote the production of ATP by increasing the activity and content of respiratory chain enzyme complex in liver mitochondria.3.Huayu Qutan recipe may prevent AS by regulating genes and proteins related to mitochondrial energy metabolism and improving the lipid deposition in the liver of as rabbits.