OBJECTIVE To express recombinant protein PBP-ApoA1 by fusion of P-selectin banding peptide(PBP)and apoli-poprotein(ApoA1)by Escherichia coli,and PBP-ApoA1 was applied to further prepare a recombinant high-density lipoprotein(HDL)loading with curcumin(Cur),named PA-rHDL-Cur,for the effective treatment of atherosclerosis(AS)by targeting to activated plate-lets.METHODS The soluble expression of PBP-ApoA1 was achieved using a co-expression strategy with glutathione S-transferase(GST)tag.The purified PBP-ApoA1,phospholipid and cholesterol were encapsulated with Cur to prepare PA-rHDL-Cur by thin-film hydration method.The physicochemical properties of PA-rHDL-Cur were characterized by particle size analyzer and UV spectrophotom-eter,while the release stability was evaluated using dialysis method.Cell viability and cellular uptake efficiency of PA-rHDL-Cur were assessed in vitro.Platelet adhesion experiments were conducted to confirm the targeting ability of PA-rHDL towards activated platelets.Furthermore,the antioxidant activity,cholesterol efflux effect,and reduction in oxidized high-density lipoprotein uptake capacity of RAW264.7 macrophages treated with PA-rHDL-Cur were investigated.RESULTS The yield of PBP-ApoA1 obtained by shake flask fermentation and purification was 1.3 g·L-1.The resulting PA-rHDL-Cur exhibited uniform particle size with an average diameter of(165.3±29.6)nm and the Zeta potential of(-2.19±1.28)mV.The biocompatibility of this drug delivey system was satisfactory.In vitro cell experiments demonstrated that PA-rHDL-Cur effectively targeted atherosclerotic lesions,releasing curcumin to reduce oxidative stress within foam cells at the lesion site,significantly enhancing the bioavailability of Cur.Additionally,the presence of ApoA1 in PA-rHDL facilitated cholesterol efflux,thereby delaying the progression of atherosclerosis.CONCLUSION This design of biomimetic recombinant high-density lipoprotein nano-drug delivery system provides a new approach and theoretical basis for the development of novel nanocarriers against atherosclerosis.
apolipoprotein A1recombinant high-density lipoproteinfusion proteinatherosclerosistargeted drug delivery
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