The Effect And Mechanism Of Wnt5a On Cholesterol Accumulation In Raw264.7/VMSC

Objective: To investigate the effect and mechanism of Wnt5a on lipid-loadedcells.Methods: Ox-LDL was used to induce Raw264.7and VMSCs into lipid-loadedcells, The expression of Wnt5a in lipid-loaded cells was detected by Western bloting.Wnt5was instantaneously high-expressed when added recombined Wnt5a. The oil redO staining was used to observe the intracellular lipid, also the content of cellular totalcholesterol and free cholesterol were determined by HPLC. The expression of proteinWnt5a、Caveolin-1、ABCAI were measured by Western bloting. Then adenovirusrecombinantion method was used to build Raw264.7stable cell lines.Results: With the increase of the concentration of ox-LDL, the expression ofWnt5a was increased in concentration dependent manner, also with the extend ofox-LDL processing time, the expression of Wnt5a was increased in time-lineardependent manner. After treated cells with Wnt5a siRNA and ox-LDL, theintracellular lipid droplet increased, the content of TC and FC increased significantly(P<0.5) compare with the group only treated with ox-LDL; but after treated cells withrWnt5a and ox-LDL, the intracellular lipid droplet increased, but the content of TCand FC decreased significantly (P<0.5) compare with the group only treated withox-LDL. In the group of cells treated with Wnt5a siRNA and ox-LDL, the expressionof Caveolin-1and ABCAI protein obviously reduced; but in the group of treated withrWnt5a and ox-LDL, the expression of Caveolin-1and ABCAI protein increased.Conclusion:1. The expression of Wnt5a showed the dependence with the concentration and timeof ox-LDL in Raw264.7and VMSCs cells. 2. Restructured Wnt5a genes could reduce the accumulation of lipid drops andcholesterol intracellular, but knocked-out Wnt5a gene was the opposite.3. Wnt5a can reduce the accumulation of cholesterol in lipid-loaded cells throughcontrol the proteins such as Caveolin-1and ABCAI which was related to cholesteroltransport.