Explore The Buyang Huanwu Decoction For Reverse Cholesterol Transport Of Atherosclerosis

Objective:To investigate the mechanisms through which Buyang Huanwu decoction prot-ects and treats for atherosclerosis in high-fat fed ApoE-/-mice by regulating the expression of cholesterol reverse transport related proteinsMethod:(1)With the big data of traditional Chinese medicine as a means of information technology,based on the traditional Chinese medicine system pharmacology platform,the human phenotypic ontology database,explore the possible mechanism of Erchen decoction in prevention and treatment Chinese medicine。Establishing a visual multi-level network of "Buyang Huanwu decoction-Chinese Herbs-Ingredients-Critical Targets-Key Pathways" and perform cluster analysis on vital targets.(2)The reference group was given feeds,and the remaining mice were given high-fat feed for 9 weeks.After 9 weeks,the serumTC,TG,LDL-C,HDL-C and were detected by a fully automatic biochemical analyzer.Observe with oil red O staining method,and measure the ratio of red lipid droplet’s to the lumen area.Western blot was used to determine the protein expressions of LXRα,LXRβ in aorta and liver and PPARy,ABCA1,p38,ERK1/2 and ApoA-I in liver;Realtime PCR was used to determine liver LXRα,LXRβ mRNA expression.Male ApoE-/-mice were randomly divided into a control group,a model group,a Buyang Huanwu decoction group,and an atorvastatin group,with 10 mice in each group.The control group was given feeds,and the remaining mice were given high-fat feed for 9 weeks.Biochemical analysis was performed to measure the serum levels of lipids including TC,TG,HDL-C and LDLC.Oil Red O staining was performed on aortic sections to evaluate the lipid accumulation.Western blot analysis was performed to analyze the protein expression levels of PPARy,ABCA1,p38,ERK1/2 and ApoA-I in liver,LXRa and LXRβ in both the aorta and liver tissue.Real-time PCR analysis was performed to analyze the mRNA expression levels of LXRa and LXRβ in liver.Result:(1)The 140 compounds of Buyang Huanwu decoction were analyzed through network pharmacology,and 174 core targets were screened closely related to atherosclerosis.Buyang Huanwu decoction may be involved in regulating inflammation and immune-related pathways including cancer,hepatitis B,PI3K-Akt pathway and T cell receptor signaling pathway.Cluster analysis of core targeted network interactions revealed that core targets are involved in biological processes such as cholesterol efflux,cholesterol homeostasis,cholesterol reverse transport and apoptosis.Among them,the core targets p3 8 and ApoA-I were confirmed participate in the treatment of atherosclerosis by Buyang Huanwu decoction though subsequent experiments,while Buyang Huanwu decoction had no significant effect on ERK1/2.(1)Oil Red O staining revealed that the in aortic sinus of the model group had fewer redstained lipids compared with that of the control group(p<0.001).Compared with the model group,the atorvastatin group and the Buyang Huanwu decoction group can significantly reduce the proportion of lipid droplets in the aorta(p<0.01).Moreover,the mice in the model group had higher serum TC,TG,LDL-C and HDL levels(p<0.001).Compared with the model group,the atorvastatin group and the Buyang Huanwu decoction group can reduce the levels of TC,TG,and LDL-C in ApoE-/-mice(p<0.001).Compared with the control group,the proteins expressions of LXRa in aortic sinuses and LXRα,LXRβ,ABCA1,phosphorylated p38,ApoA-I in liver in the model group were significantly reduced(p<0.05,p<0.01,p<0.001).However,the protein expression of PPARy,phosphorylated ERK1/2 and mRNA expression levels of LXRα,LXRβ in the liver were not statistically significant(p>0.05).Compared with the model group,the atorvastatin and the Buyang Huanwu decoction were able to effectively up-regulates the protein expression of LXRα and ABCA1 in the liver(p<0.05,p<0.01).In addition,the Buyang Huanwu decoction can also increase the protein expression levels of LXRα,LXRβ in the aorta,and the protein expression levels of LXRβ,ApoA-I,phosphorylated p38 in the liver(p<0.05,p<0.01,p<0.001).At the same time,blocking of phosphorylated p38 and LXRβ eliminated the upregulation of ApoA-I induced by BHD in the liver of C57BL/6J mice(p<0.01),but did not ABCA1(p>0.05).Conclusion:This study demonstrated that in ApoE-/-mice,Buyang Huanwu decoction treatment significantly reduced the lipid droplets in the blood and lipid accumulation in the aortic sinus.Compared with atorvastatin,Buyang Huanwu decoction affect proteins of regulate cholesterol reverse transport easily.Besides it with the characteristics of multi-target for prevention and treatment atherosclerosis.The study blocked p38 and LXR for the first time,and further proved that BHD through the LXRα/ABCA1 pathway and activation of phosphorylated p38,LXRβincreases the expression of ApoA-I to promote liver cholesterol reverse transport and thus regulate the occurrence and development of AS.