| Objective: 1.To observe the effect of Danshen Injection on blood lipid in hyperlipidemia mice induced by hyperlipidemia emulsion.2.Screen the target and mechanism of Danshen injection in the treatment of hyperlipidemia by network pharmacology.3.Based on PPARγ expression of key proteins in signal pathway,the prediction results of network pharmacology were preliminarily verified to explore the mechanism of Salvia miltiorrhiza injection in improving hyperlipidemia.Methods: 1.The hyperlipidemia mouse model was established by intragastric administration of high-fat emulsion,and the successful mice were randomly divided into model group,fenofibrate group(26 mg/kg),high-dose Salvia miltiorrhiza injection group(5 m L/kg)and low-dose Salvia miltiorrhiza injection group(2.5 m L/kg).The model group and each administration group continued to make the model by intragastric administration of high-fat emulsion.Mice in each group were intragastric administration of normal saline or high-fat emulsion in the morning and intraperitoneal injection of normal saline or corresponding drugs in the afternoon for 21 days.Mice in each group were intragastric administration of water or fat emulsion in the morning and intraperitoneal injection of water or corresponding drugs in the afternoon for21 days.The blood lipid indexes of mice in each group were measured at the end of each week,and the blood lipid indexes were measured at the end of each week;The liver was separated,weighed and the liver coefficient was calculated;He staining was used to observe the pathomorphological changes of liver tissue.2.Network pharmacological analysis based on the above experimental research.The effective chemical components of Salvia miltiorrhiza injection were obtained through literature research,and the action targets of the effective components of Salvia miltiorrhiza injection were obtained by using Swiss target prediction and pharmmapper database.The disease targets of hyperlipidemia were collected by searching five disease target databases: drug bank,OMIM,disgenet,gene cards and therapeutic target database.The intersection of disease target and component target is obtained by using Venny platform to obtain the potential therapeutic target of Danshen injection in the treatment of hyperlipidemia.The obtained intersection targets were introduced into the string network platform to construct PPI,and the obtained results were introduced into Cytoscape 3.8.2.The cytohubba plug-in was used to screen the core gene,and MCC topology algorithm was used for visualization analysis to determine the core lipidlowering target of Danshen injection.David platform is used for go function and KEGG pathway enrichment analysis,and visual analysis is carried out through omicshare.3.Carry out experimental verification according to the results of network pharmacology,and take the serum and liver of the above experimental animals as the research object.ELISA was used to detect serum HMG-Co A,LCAT,LPL and HL,CYP7A1 and PPARγ in liver tissue Level.Western blot was used to detect PPARγ,LXRα、 ABCA1 and CYP7A1 protein expression in mouse liver tissue.Results: 1.Study on the lipid-lowetring effect of Danshen Injection on hyperlipidemia model mice: compared wih the blank group,the contents of TG,CHO,LDL-C in serum and MDA in liver tissue in the model group increased significantly(P < 0.05 or P < 0.01)and the contents of SOD and GSH-Px in liver tissue decreased significantly(P < 0.05);Compared with the model group,the contents of CHO and LDL-C in each treatment group decreased significantly,and the content of TG in fenofibrate group decreased significantly(P < 0.05 or P < 0.01).The contents of SOD and MDA in Salvia miltiorrhiza injection group(P < 0.05)and low-dose Salvia miltiorrhiza injection group were significantly decreased(P < 0.05);The results of liver histopathology showed that each administration group could improve the lipid accumulation in liver tissue.2.Network pharmacology study of Salvia miltiorrhiza injection in the treatment of hyperlipidemia: there are 108 potential targets of Salvia miltiorrhiza injection in the treatment of hyperlipidemia,including RXRα、PPARγ、RXRγ、RARα、TNF and other 10 target proteins may be the key targets for the treatment of diseases.According to the analysis of go and KEGG,the lipid-lowering mechanism may be related to signal pathways such as glucose and lipid metabolism and inflammatory response.3.Study on the mechanism of Salvia miltiorrhiza injection on lipidlowering in hyperlipidemia model mice: compared with the blank group,in the model group,the content of HMG-Co A in serum of model mice increased significantly(P < 0.05),LCAT,LPL in serum and PPARγ,HL,CYP7A1 in liver tissue were significantly reduced,the protein expression levels of liver PPARγ,LXRα,ABCA1 and CYP7A1 decreased significantly(P < 0.05 or P < 0.01);Compared with the model group,the levels of LCAT,LPL,PPARγ,HL and CYP7A1 increased significantly(P < 0.05 or P < 0.01).Salvia miltiorrhiza injection group could significantly increase CYP7A1,LCAT and PPAR γ,and the content of HMG-Co A in high dose group could be significantly reduced(P< 0.05).PPARγ and CYP7A1 protein expression of each administration group increased significantly,and LXRα,ABCA1 in high dose group increased significantly(P < 0.05).WB results suggest that Danshen injection can up regulate the protein expression levels of ABCA1,CYP7A1,PPARγ and LXRα(P < 0.05).Conclusion: 1.Salvia miltiorrhiza injection can effectively improve the blood lipid level of hyperlipidemia model mice induced by hyperlipidemia emulsion,reduce the level of oxidative stress and improve hepatocyte injury.2.Network pharmacology research shows that Danshen injection plays a lipidlowering role through multi-component,multi-target and multi-channel,and its mechanism may be related to signal pathways such as glucose and lipid metabolism and inflammatory response.3.Danshen injection may activate PPARγ-LXRα-CYP7A1/ABCA1 signaling pathway to reduce cholesterol synthesis,promote cholesterol transport and metabolism to play a role in lipid-lowering. |
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