Effects Of Berberine On Macrophage Autophagy During Arteriosclerosis Process

Objective: After treatment with oxLDL,macrophages containing large amounts of lipids are foam cells,detect the changes of autophagy function of macrophages under the condition of atherosclerosis,and the regulatory effect of berberine and its molecular mechanism.Methods: The mouse macrophage J774 A.1 was cultured to construct the foam cell model.The cells were randomly divided into 5 groups.normal control group,25 ng / ?l oxLDL group,50 ng / ?l oxLDL group,75 ng / ?l oxLDL group and Oil red O was used to detect the lipid content of the cells.CD36 was used as a marker protein for lipid metabolism in macrophages.The protein was collected and the changes of CD36 were detected by Westren-blot.To investigate the effect of berberine on the mRNA expression of autophagy-related proteins Beclin-1,LAMP,P53 and Bcl-2 in macrophages,the cells were randomly divided into five groups.normal control group,6.25?M BBR group,12.5?M BBR group,25?M BBR group and 50?M BBR group.Total RNA was extracted and reversely transcribed into cDNA Real-time PCR was used to detect the mRNA expression of autophagy-related protein Beclin-1,LAMP,P53 and Bcl-2.To further investigate the effect of BBR on lipid accumulation and autophagic function of foam cells,the cells were randomly divided into 4 groups: normal control group,75 ng / ?loxLDL group,75 ng / ?loxLDL + 25 ?MBBR,75 ng / ?loxLDL + 25?M BBR + chloroquine 30?M(CQ)group.Western Blot was used to detect the changes of autophagy markers P62,LC3 and CD36.Results: 1.Oil red O staining results show thatl ipid content exceeds 50% of the cell volume at 50 ng / ?l,75 ng / ?l,100 ng / ?l oxLD concentration.Westren-blot results showed that CD36 was significantly elevated at oxLDL concentrations of 25 ng / ?l,50 ng / ?l and 75 ng / ?l,but not at 100 ng / ?l.Foam cells showed successful modeling.2.The mRNA level of Beclin-1 increased most significantly at the concentration of BBR25?M.The LAMP increased obviously at12.5?M,while the mRNA expression of P53 was significantly increased at25?M BBR.The mRNA expression level of Bcl-2 was significantly increased at 6.25?M,12.5?M,25?M and 50?M BBR.3.LC3? / LC3?had a slight increase after adding BBR,P62 slightly decreased,after adding BBR,CD36 decreased.Conclusions: BBR can enhance the autophagy of macrophages and promote the lipid metabolism of macrophages and reduce the formation of foam cells by enhancing autophagy.