| 朱云峰,覃艳,曹萌,等.黄芪多糖对脂肪组织巨噬细胞活化的影响及作用机制[J].安徽医药,2021,25(12):2350-2354. |
| 黄芪多糖对脂肪组织巨噬细胞活化的影响及作用机制 |
| Effect of Astragalus polysaccharides on adipose tissue macrophage activation and its mechanism |
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| DOI：10.3969/j.issn.1009-6469.2021.12.005 |
| 中文关键词: 黄芪 脂肪组织 胰岛素抵抗 黄芪多糖 肿瘤坏死因子 -α 巨噬细胞 号通， |
| 英文关键词: Astragalus membranaceus Adipose tissue Insulin resistance Astragalus polysaccharide TNF-α Macrophage |
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| 中文摘要: |
| 目的探讨黄芪多糖( APS)对脂肪组织巨噬细胞活化的影响及作用机制。方法 2018年 11月至 2019年 5月，通过 0.4 μm孔径大小的 Transwell小室共培养方式，将巨噬细胞种于上室，将脂肪细胞种于下室。实验分为三组，空白对照组( NC组)、肿瘤坏死因子 -α(TNF-α)诱导干预组( TNF-α组)和 APS干预组( APS+TNF-α组)酶联免疫吸附测定( ELISA)检测细胞培养上清液中白细胞介素 -6(IL-6)、单核细胞趋化蛋白 -1(MCP-1)水平，实时荧光定量录聚合酶链反应( qRT-PCR)检测巨噬细胞诱生型一氧化氮合酶( iNOS)和 TNF-α mRNA表达水平，蛋白质印迹法( Western blotting)检测 iNOS和磷酸化 AMP活化蛋白激酶 α1(p-AMPKα1)和沉默信息调节蛋白 1(SIRT1)水平。同时，采用 8 μm孔径大小的 Transwell小室共培养方式，结晶紫染色观察巨噬细胞的趋化情况。结果与 NC组比较， TNF-α组细胞培养上清液中 IL-6和 MCP-1水平显著升高( P<0.05)，巨噬细胞中 TNF-α mRNA、iNOS mRNA和蛋白水平显著升高( P<0.05)p-AMPKα1和 SIRT1蛋白水平显著降低( P<0.05)趋化巨噬细逆转，胞数显著升高( P<0.05)其中 NC组 iNOS mRNA和蛋白水平分别为，(1.00 ± 0.17)、(0.43 ± 0.05)TNF-α组分别为(3，.57±0.31)、(1.18 ± 0.16)。与 TNF-α比较， APS+TNF-α组细胞培养上清液中 IL-6和 MCP-1水平显著降P<0.05)，巨噬细胞中 TNF-α组，低(mRNA、iNOS mRNA和蛋白水平显著降低( P<0.05)p-AMPKα1和 SIRT1蛋白水平显著升高( P<0.05)趋化巨噬细胞数显著降低( P<0.05)，其中 TNF-α组 iNOS mRNA和蛋白水平(，3.57±0.31)、(1.18 ± 0.16)APS+TNF-α组分别为(，1.87 ± 0.22)、(0.62 ± 0.08)。结论 APS可抑制巨噬细胞的活化，其作用机制与激活 AMPK/SIRT1信路有关。 |
| 英文摘要: |
| Objective To investigate the effect of Astragalus polysaccharide (APS) on the activation of adipose tissue macrophages and its mechanism.Methods From November 2018 to May 2019, macrophages were seeded in the upper chamber and fat cells werecultured in the lower chamber by a 0.4 μm pore size Transwell chamber co-culture method . The experiment was assigned into three groups: blank control group (NC group), tumor necrosis factor-α (TNF-α) induction intervention group (TNF-α group) and APS interven. tion group (APS+TNF-α group). The levels of interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in the cell culture su. pernatant were detected by enzyme-linked immunosorbent assay (ELISA). Real-time quantitative PCR (qRT-PCR) was used to detect the mRNA expression level of inducible nitric oxide synthase (iNOS) and TNF-α in macrophages. Western blotting was used to detect iNOS and phospho-adenosine monophosphate activated protein kinase alpha 1 (p-AMPKα1) and silent information regulator protein 1(SIRT1) levels. At the same time, the chemotaxis of macrophages was observed by crystal violet staining using a Transwell chamber co-culture method with a pore size of 8 μm.Results Compared with NC group, the levels of IL-6 and MCP-1 in the culture supernatant of TNF-α group were significantly increased (P<0.05), and the mRNA levels of TNF-α and iNOS and the protein level of iNOS were signif. icantly increased (P<0.05), and the levels of p-AMPKα1 and SIRT1 protein were significantly reduced (P<0.05), and the number of chemotactic macrophages was increased (P<0.05). The level of iNOS mRNA and protein in NC group was (1.00±0.17) and (0.43±0.05), respectively, and those in TNF-α group was (3.57±0.31) and (1.18±0.16), respectively. Compared with TNF-α group, the levels of IL-6 and MCP-1 in APS+TNF-α group were significantly decreased (P<0.05), and the mRNA levels of TNF-α and iNOS and the protein lev. el of iNOS were significantly decreased (P<0.05), and the levels of p-AMPKα1 and SIRT1 protein were significantly increased (P< 0.05), and the number of chemotactic macrophages was reduced (P<0.05). The level of iNOS mRNA and protein in TNF-α group were (3.57±0.31) and (1.18±0.16), respectively, those in APS+TNF-α group were (1.87±0.22) and (0.62±0.08), respectively.Conclusion APS can inhibit the activation of macrophages, and its mechanism is related to the activation of AMPK/SIRT1 signaling pathway. |
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