Abstract: OBJECTIVE: To investigate the efficacy of Bushen Kangshuai(BS-KS) tablet on autophagy and polarization in mouse macrophage RAW 264.7.MEYHODS: Macrophage autophagy was induced by oxidized low-density lipoprotein(100 μg/m L). To detect the levels of autophagy, macrophage were transfected with double fluorescence LC3 autophagy adenovirus, then the numbers of autophagosomes and autophagic lysosomes were asessed by confocal microscopy. The autophagy related proteins expression of PI3 K, Akt, phospho-m Akt(p-Akt) and m TOR, phospho-m TOR([p-TOR), p62,microtubule-associated protein 1(LC3-Ⅱ)were determined by western blotting. The macrophage polarization model was induced by lipopolysaccharide(1 μg/m L). The m RNA levels of i NOS, CD86(M1 macrophages marker molecules), and CD206, Arg-1(M2 macrophages marker molecules) were detected by real-time quantitative PCR. The concentration of cytokines TNF-α and IL-10 was determined by enzyme-linked immunosorbent assay. The protein expression of nuclear proteins PPAR-γ, NF-κB,and cytoplasmic protein IKB α was determined by western blotting.RESULTS: The expression of the autophagy-related protein LC3-Ⅱ was increased and the expression of p62 was decreased in the BS-KS intervention group. The protein expression of PI3 K, p-Akt, and p-m TOR was also reduced. BS-KS also inhibited the m RNA expression of i NOS and CD86 on M2 macrophage, but promoted the expression of CD206 and Arg-1 on M2 macrophage. With respect to the regulation of inflammatory factors, BS-KS could inhibit the secretion of pro-inflammatory TNF-α and promote the secretion of anti-inflammatory IL-10. It also inhibited the protein expression of IKB-α and NF-κB, and promoted the expression of nuclear protein PPAR-γ.CONCLUSION: We believe that BS-KS promotes macrophage autophagy by increasing the level of autophagy protein and inhibiting the PI3 K/Akt/m TOR signaling pathway. Furthermore, BS-KS seems to inhibit macrophage M1 polarization and promote M2 polarization via the PPAR gamma/NF-κB signaling pathway, thus playing an inhibitory role in atherosclerosis.

Key words: Atherosclerosis, Macrophages, Autophagy, Lipoproteins,LDL, Lipopolysaccharides, Signal transduction, Bushen Kangshuai tablets