Effect Of PPARδ On Lipid Accumulation In Macrophages Induced By OA And Ox-LDL

Objective:The foam cell model was constructed by incubating mouse-derived RAW264.7macrophages with OA and ox-LDL to clarify the effect of OA and ox-LDL on PAT family proteins and the expression of PPARδ,so as to clarify the effect of PPARδ on PAT family proteins and provide a new therapeutic basis for anti-atherosclerosis.At the same time,by studying the effect of PPARδ on lipid accumulation,it provides a possible new strategy for the treatment of atherosclerosis.Methods:1.RAW264.7 cells were incubated with different lipid components ox-LDL or OA for 24 h,and the effects of ox-LDL and OA on the expression of ADRP and TIP47 protein were detected by Western Blot.2.RAW264.7 cells were incubated with different concentrations of ox-LDL or OA for 24 h,and the expression of PPARδ m RNA was detected by real-time PCR.3.RAW264.7 cells were incubated with ox-LDL and OA for 24 h,and the expression level of PPARδ m RNA was detected by real-time PCR.4.RAW264.7 cells were treated with different concentrations of PPARδ agonist GW501516(10n M,100 n M)and PPARδ inhibitor GSK0660 for 24 h,and the expression level of CD36 m RNA was detected by real-time PCR.5.RAW264.7 cells were pretreated with different concentrations of PPARδ agonist GW501516(10n M,100 n M)for 1h,and then incubated with ox-LDL or OA for 24 h.The expression levels of ADRP and TIP47 m RNA were detected by real-time PCR.6.RAW264.7 cells were pretreated with 100 n M PPARδ agonist GW501516 and PPARδ inhibitor GSK0660 for 1 h,and then incubated with ox-LDL or OA for 24 h.The expression of ADRP was detected by real-time PCR and Western Blot.Results:1.Compared with the control group,the protein expression levels of ADRP in OA group and ox-LDL group were increased(P < 0.05).There was no significant difference in the protein expression level of TIP47.2.With the increase of OA concentration,the expression level of PPARδ m RNA in macrophages increased gradually,and the expression of PPARδ m RNA increased most significantly when OA concentration was 200μM(P < 0.05).3.With the increase of ox-LDL concentration,the expression level of PPARδm RNA in macrophages increased gradually,and the expression of PPARδ m RNA increased most significantly when the concentration of ox-LDL was 50μg / m L(P <0.05).4.The expression level of PPARδ m RNA was increased in 200μM OA group,50μg/ m L ox-LDL group and 200μM OA + 50μg / m L ox-LDL group(P < 0.05),and the expression of PPARδ m RNA was increased most significantly after RAW264.7 cells were incubated with OA combined with ox-LDL(P < 0.05).There was no significant difference in the expression of PPARδ m RNA between OA group and ox-LDL group.5.PPARδ agonist GW501516 up-regulated the expression of CD36 m RNA in RAW264.7 macrophages in a concentration-dependent manner,and the expression of CD36 m RNA increased most significantly when treated with 100 n M GW501516(P <0.05),while the addition of PPARδ inhibitor GSK0660 down-regulated the expression of CD36 m RNA(P < 0.05).6.PPARδ agonist GW501516 could up-regulate the expression of ADRP m RNA in RAW264.7 macrophage cells induced by OA or ox-LDL in a concentration-dependent manner,and the expression of ADRP m RNA increased most significantly when 100 n M GW501516 intervened(P < 0.05),but the expression level of TIP47 did not change significantly before and after treatment.7.PPARδ agonist GW501516 could up-regulate the expression of ADRP m RNA in RAW264.7 cells induced by OA or ox-LDL,and the addition of PPARδ inhibitor GSK0660 could reverse this result(P < 0.05),which was also verified in the detection of ADRP protein expression level.Conclusion:1.OA and ox-LDL up-regulated the expression of ADRP in RAW264.7macrophages,but had no significant effect on the expression of TIP47.2.OA and ox-LDL up-regulated the expression of PPARδ in RAW264.7 cells and had a synergistic effect.3.PPARδ agonist GW501516 up-regulated the expression of CD36 in RAW264.7cells,while PPARδ inhibitor GSK0660 down-regulated the expression of CD36.4.PPARδ agonist GW501516 up-regulated the expression of ADRP in RAW264.7cells induced by OA and ox-LDL,but had no significant effect on the expression of TIP47,while PPARδ inhibitor GSK0660 down-regulated the expression of ADRP,suggesting that PPARδ may be involved in lipid accumulation.