Prevention Of Atherosclerosis In High Fat Fed Apoe^-/- Mice With LRRC8A Chloride Channel As Target

Objective : To observe the effect of LRRC8 A on atherosclerosis in high fat fed ApoE^-/-mice and the effect of volume regulation of chloride channel inhibitors 9-AC and NPPB on atherosclerosis.To observe the effects of RAW264.7 cell foaming process In the LRRC8 A changes and interference with LRRC8 A RAW264.7 cells after the bubble situation.The mechanism of LRRC8 A on the foaming of RAW264.7 cells was discussed.Methods : ApoE-/-mice were fed with high-fat diet for 12 weeks.During the injection of solvent-regulated chloride channel inhibitors?9-AC and NPPB?,the size of the thoracic aortic plaques was observed by oil red O and HE staining.The macrophages of the mice were examined by Western blot,and the expression of LRRC8 A protein was detected by Western blot.The cells treated with ox-LDL were treated with isotonic,hypotonic culture medium and volume-regulated chloride channel inhibitor?9-AC and NPPB?.The low permeability of RAW264.7 cells was detected by Western blot.,9-AC,NPPB on LRRC8 A and CD36 protein expression.Oil red O staining was used to observe the degree of cell foaming.Transient transfection of si LRRC8 A,interference LRRC8 A protein expression,plus the above treatment factors oil red O staining to observe cell foam.To confirm whether LRRC8 A affected foaming of RAW64.7 macrophages by influencing CD36 and Arf-6 pathways,the interaction between CD36 and Arf-6 was observed by immunoprecipitation.Results:The accumulation of placental aorta in ApoE^-/-mice was increased after 12 weeks of high fat diet,while NPPB and 9-AC inhibited the plaque formation.The expression of LRRC8 A in the peritoneal macrophages of the model group was increased?4.251 ± 0.507 vs WT1.000 ± 0.0?.The volume regulation of the chloride channel inhibitor NPPB and 9-AC inhibited the expression of LRRC8A?1.179 ± 0.251,1.091 ± 0.138?.In the RAW264.7 cell foaming process,the hypotonic effect increased LRRC8 A protein expression?1.388 ± 0.171?,and the solvent-regulated chloride channel inhibitor 9-AC and NPPB inhibited the expression of low-permeability LRRC8A?0.514 ± 0.106,0.765 ±0.079?.Oil red O results suggest that low permeability can induce RAW264.7 cell foaming,solvent-mediated chloride channel inhibitor9-AC inhibits cell foam,and si LRRC8 A inhibits ox-LDL-induced RAW264.7 cell foam.Hypoglycemia increased CD36 protein expression?1.177 ± 0.058?and si LRRC8 A inhibited CD36 expression?0.5586 ±0.112?.The results of immunoprecipitation of CD36 and Arf-6 showed that the low expression of Arf-6 induced by ox-LDL-induced RAW264.7cell was up-regulated and si LRRC8 A was up-regulated.Conclusion:.1.LRRC8 A can mediate high fat fed ApoE^-/-mouse atherosclerosis progression.Volume-regulated channel inhibitors9-AC and NPPB inhibit mouse atherosclerosis.The expression of LRRC8 A was increased in the process of bubbling induced by ox-LDL,and the interference of LRRC8 A inhibited the oxidation of RAW264.7cells induced by ox-LDL.2.LRRC8 A may accelerate the foaming of RAW264.7 cells by promoting binding of CD36 and Arf-6.