Preliminary Study On The Intervention Of Xiong Shiwei Wendan Decoction On Reverse Cholesterol Transport In Foam Cells

Objective: to explore whether Xiong Shiwei Wendan decoction can promote cholesterol outflow from foam cells through reverse cholesterol transport pathway,so as to play the role of anti-AS for the prevention and treatment of cardiovascular related diseases.Verify and enrich the scientific connotation of "phlegm-heat" pathogenesis of chest arthralgia and heartache in traditional Chinese medicine from the perspective of RCT,and provide new ideas for the prevention and treatment of As with traditional Chinese medicine.Methods: 1.Xiong Shiwei Wendan decoction was prepared by centrifugation,high temperature and high pressure sterilization,aseptic membrane filtration and other steps to prepare Xiong Shiwei Wendan decoction.2.Mouse RAW264.7-derived macrophages were induced by foaming with a final concentration of 25 μ g / mg of OX-LDL.3.The concentrated preparation of Xiong Shiwei Wendan decoction was diluted into different concentration gradients,and the cell survival rate was determined by CCK-8 cell proliferation test.Then the foam cells were intervened with selected traditional Chinese medicine reagents with different concentration gradients.The accumulation of intracellular lipid droplets was observed by oil red O staining,and the levels of mi R-33 m RNA and ABCA1 m RNA were detected by Real-time PCR,so as to explore the effect of Xiong Shiwei Wendan decoction on promoting RCT anti-AS.Results: 1.RAW264.7 macrophages were induced by different concentrations of OX-LDL.By detecting the contents of mi R-33 and ABCA1 m RNA in the cells,we finally selected the concentration of 25.0 μ g / mg to construct the macrophage foam cell model.2.The suitable concentration of Xiong Shiwei Wendan decoction was screened out by CCK-8 experiment.We found that when the drug concentration was higher than 5mg/ml,the cell survival rate was less than 50%,so three concentration gradients of 1.25mg/ml,2.5mg/ml and 5mg/ml were selected for further experiment.3.Oil red O staining was used to observe the accumulation of lipid droplets in foam cells under the intervention of different concentrations of traditional Chinese medicine.The results showed that Xiong Shiwei Wendan decoction could reduce the accumulation of lipid droplets in foam cells in a certain concentration range,but there was no significant difference between 2.5mg/ml and 5mg/ml groups,but it was significantly better than that in 1.25mg/ml groups.4.Through Real-time PCR detection of ABCA1 and mi R-33 m RNA expression levels under the intervention of different concentrations of traditional Chinese medicine,we found that Xiong Shiwei Wendan decoction could reduce the level of mi R-33 m RNA and increase the level of ABCA1 m RNA in the concentration range of 2.5mg/ml and 5.0mg/ml,thus exerting the effect of regulating lipid and promoting reverse cholesterol transport.Conclusion: Xiong Shiwei Wendan decoction can promote cholesterol outflow from foam cells in a certain concentration range,thus reducing the accumulation of intracellular lipid droplets,which may be achieved by down-regulating mi R-33 and up-regulating the level of ABCA1.The purpose of this study is to provide a new idea for the prevention and treatment of atherosclerosis by reverse cholesterol transport of traditional Chinese medicine.