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目的研究黄芪多糖(astragalus polysaccharide,APS)对人单核细胞THP-1前蛋白转化酶枯草溶菌素9(proprotein convertase subtilisin/kexin type 9,PCSK9)表达的影响,并初步探讨其机制。方法利用佛波酯诱导THP-1细胞分化为THP-1源性巨噬细胞,采用CCK-8法检测不同浓度APS对THP-1源性巨噬细胞增殖活性的影响,确定APS作用于细胞的最适浓度。将THP-1源性巨噬细胞分为5组:低密度脂蛋白(low density lipoprotein,LDL)组(30μg/ml)、LDL(30μg/ml)+APS(400μg/ml)组、LDL(30μg/ml)+过氧化物酶体增殖物激活受体β(peroxisome proliferatoractivated receptorβ/δ,PPAR-β/δ)激动剂GW0742(1μmol)组、LDL(30μg/ml)+PPAR-β/δ抑制剂GSK0660(1μmol)+APS(400μg/ml)组和对照组(不加药物),加药处理24 h后,分别采用RT-PCR及Western blot法检测各组细胞PCSK9、低密度脂蛋白受体(low density lipoprotein receptor,LDLR)、PPAR-β/δ基因及蛋白的表达量。结果APS作用于THP-1源性巨噬细胞的最适浓度为400μg/ml。LDL组细胞PCSK9基因及蛋白的表达量均较对照组明显升高(P<0.01),LDL+APS和LDL+GW0742组较LDL组明显降低(P<0.01),LDL+GSK0660+APS组较LDL+APS和LDL+GW0742组明显升高(P<0.01)。LDL、LDL+APS、LDL+GW0742和LDL+GSK0660+APS组细胞LDLR基因的表达量较对照组均明显降低(P<0.01);LDL组细胞LDLR蛋白的表达量较对照组降低(P<0.01),LDL+APS和LDL+GW0742组较LDL组升高(P<0.01),LDL+GSK0660+APS组较LDL+APS和LDL+GW0742组明显降低(P<0.01)。LDL+APS和LDL+GW0742组细胞PPAR-β/δ基因及蛋白的表达量较对照组均显著升高(P<0.01),LDL+GSK0660+APS组较LDL+APS和LDL+GW0742组明显降低(P<0.01)。结论 APS可降低THP-1源性巨噬细胞PCSK9的表达,且PPAR-β/δ可能是其作用通路之一。
Objective To investigate the effect of astragalus polysaccharide (APS) on the expression of proprotein convertase subtilisin / kexin type 9 (THP-1) in human monocytes. Methods THP-1 cells were induced to differentiate into THP-1-derived macrophages by phorbol ester. The effects of different concentrations of APS on the proliferation of THP-1-derived macrophages were determined by CCK-8 assay. Optimum concentration. THP-1-derived macrophages were divided into 5 groups: low density lipoprotein (LDL) group (30μg / ml), LDL (30μg / ml) / ml) + peroxisome proliferator-activated receptorβ / δ (PPAR-β / δ) agonist GW0742 (1μmol) After treatment with GSK0660 (1μmol) + APS (400μg / ml) and control group (without drug) for 24 h, the expression of PCSK9 and low density lipoprotein receptor low density lipoprotein receptor (LDLR), PPAR-β / δ gene and protein expression. Results The optimal concentration of APS for THP-1-derived macrophages was 400 μg / ml. LDL + APS and LDL + GW0742 groups were significantly lower than LDL group (P <0.01), LDL + GSK0660 + APS group than LDL group were significantly higher than the control group (P <0.01) + APS and LDL + GW0742 group was significantly higher (P <0.01). Compared with control group, the expression of LDLR gene in LDL, LDL + APS, LDL + GW0742 and LDL + GSK0660 + APS group was significantly lower than that in control group (P <0.01); the LDLR protein expression in LDL group was lower than that in control group (P <0.01). LDL + GSK0660 + APS group was significantly lower than LDL + APS and LDL + GW0742 group (P <0.01). Compared with control group, the expression of PPAR-β / δ gene and protein in LDL + APS and LDL + GW0742 groups were significantly increased (P <0.01), LDL + GSK0660 + APS group was significantly lower than LDL + APS and LDL + GW0742 group (P <0.01). Conclusion APS can reduce the expression of PCSK9 in THP-1-derived macrophages, and PPAR-β / δ may be one of its pathways.