Effect Of CLIC1 On Macrophage Lipid Accumulation And Inflammation And Intervention Of Gypenosides

Objective To investigate the role of chloride ion channel protein 1（CLIC1）in lipid accumulation and inflammation of macrophages and elucidate the molecular mechanism of anti-atherosclerosis of gypenosides from the perspective of CLIC1regulation.Methods Twenty four ApoE^-/-mice were randomly divided into four groups,namely normal group,model group,low dose group of gypenosides,high dose group of gypenosides,6 rats in each group.After 8 weeks of high-fat diet,the blood lipid levels of the animals were analyzed by enzymatic method,the area of atherosclerotic plaque was analyzed by H&E staining,the expression of CLIC1 in plaques was observed by immunohistochemistry,the expression of TLR4,My D88,NF-κB and CLIC1 by western blot.At the same time,we cultured RAW264.7 cells in vitro and were treated with 50μg/m L ox-LDL for 24 h and treated with IAA94/CLIC1 si RNA or gypenosides,the contents of intracellular TC and FC were detected by chemical method,the intracellular lipid accumulation was detected by oil red O,the expression of CLIC1,CD36,IL-6,TNF-αwas detected by RT-PCR and western blotting,the intracellular chloride concentration was detected by MQAE,the translocation of CLIC1 membrane was detected by immunofluorescence.Analyzing the role of CLIC1 in macrophage lipid accumulation and the effect of gypenosides on CLIC1 expression and membrane translocation.Results The level of blood lipid and the area of aortic plaques were significantly increased in AS mice,the expression of CLIC1 in plaques was significantly increased,and the expression of TLR4,My D88,NF-κB were significantly increased.After intervention with 200 and 400 mg/kg of gypenosides,the level of blood lipid in mice was significantly reduced,and the area of aortic plaque in mice was significantly smaller,the expression of CLIC1 in plaque was significantly lower,the expression of TLR4,My D88,NF-κB were significantly decreased and showed a dose-dependent.After RAW264.7 cells treated with 50μg/m L ox-LDL for 24 h,the intracellular contents of TC and FC were significantly increased,lipid accumulation was significantly increased,the expression of CLIC1,CD36,IL-6 and TNF-αwas significantly increased,and the intracellular chloride concentration was significantly increased.After intervention with IAA94,CLIC1 si RNA and different concentrations of gypenosides,intracellular TC and FC contents was significantly reduced,lipid accumulation and CLIC1,CD36,IL-6,TNF-αexpression were significantly reduced,intracellular chloride concentration was significantly reduced,the CLIC1 expression on the cell membrane was decreased.Conclusion CLIC1 is highly expressed in mouse atherosclerotic plaque and gypenosides can reduce the expression of CLIC1.CLIC1 is associated with lipid accumulation and inflammation of macrophages.After inhibition of CLIC1,the accumulation of lipid in cells is significantly reduced.The gypenosides can inhibit the lipid accumulation and inflammation of macrophages by down regulating the expression of CLIC1,thus playing an anti-atherosclerotic effect.