黄精水提物干预脂多糖诱导的巨噬细胞极化与自噬研究  

Effect of Polygonatum sibiricum aqueous extract on lipopolysaccharides induced macrophage polarization and autophagy

作　　者：李九九 汪光军 刘政祥 余锦川 梁娣 汪秋冶 陈文军[1] LI Jiu-Jiu;WANG Guang-Jun;LIU Zheng-Xiang;YU Jin-Chuan;LIANG Di;WANG Qiu-Ye;CHEN Wen-Jun(School of Public Health,Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽医科大学公共卫生学院,合肥230032

出　　处：《食品安全质量检测学报》2021年第19期7772-7777,共6页Journal of Food Safety and Quality

基　　金：青阳县九华黄精康养产业研究院安徽省院士工作站研究项目(JHHJYSGZZ19001)。

摘　　要：目的探讨黄精水提物(Polygonatum sibiricum aqueous extract,PSAE)对脂多糖(lipopolysaccharides,LPS)诱导的RAW264.7巨噬细胞M1极化与自噬的调节作用,探究其抗炎机制。方法实验分为对照组(无任何处理)、LPS组(100 ng/mLLPS刺激12 h)、PSAE低、中、高剂量组(50、100、200μg/mLPSAE预处理细胞4 h后,再加入100 ng/mL LPS刺激12 h)。细胞计数(cell counting kit-8,CCK-8)法检测细胞活性;Griess法检测细胞NO分泌量;实时荧光定量PCR检测细胞诱导型NO合酶(inducible nitric oxide synthase,iNOS)、白细胞介素-1β (interleukin-1β,IL-1β)、肿瘤坏死因子-α (tumor necrosis factor-α,TNF-α)及单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)的mRNA水平;蛋白质免疫印迹和免疫荧光法检测M1型极化标志物iNOS表达水平;蛋白质免疫印迹检测微管相关蛋白1轻链3 (microtubule-associated protein 1 light chain 3,LC3)和螯合体1 (sequestosome 1,p62)表达水平。结果与LPS组相比,PSAE可使RAW264.7细胞NO的分泌减少(P<0.05,P<0.01),PSAE也可降低M1极化相关炎症因子(iNOS、IL-1β、TNF-α、MCP-1) mRNA表达水平(P<0.05,P<0.01,P<0.001),降低M1极化标志物iNOS蛋白表达水平(P<0.05,P<0.001),同时,PSAE可降低LC3Ⅱ/Ⅰ值(P<0.01,P<0.001),促进p62蛋白表达(P<0.01),从而降低自噬水平。结论 PSAE能够抑制LPS诱导的RAW264.7细胞M1极化和自噬水平,减轻细胞炎症反应。Objective To investigate the regulatory effect of Polygonatum sibiricum aqueous extract(PSAE) on the M1 polarization and autophagy of RAW264.7 cells induced by lipopolysaccharides(LPS),and explore its anti-inflammatory effects.Methods The experiment was divided into control group(without any treatment),LPS group(treated with 100 ng/mL LPS for 12 h),PSAE low-dose,medium-dose and high-dose groups(50,100,200 μg/mL PSAE pre-treated for 4 h,respectively,and then treated with 100 ng/mL LPS for 12 h).Cell counting kit-8(CCK-8)assay was used to detect the viability of RAW264.7 cells;Griess assay was used to detect the products of NO;real-time fluorescent quantitative polymerase chain reaction method was used to detect the mRNA expression levels of inducible nitric oxide synthase(iNOS),interleukin-1β(IL-1β)tumor necrosis factor-a(TNF-α),and monocyte chemotactic protein(MCP-1);western blot and immunocellular fluorescence assay were used to detect the expression levels of M1 polarization maker,iNOS;the protein expression levels of microtubule-associated protein light chain 3(LC3) and sequestosome 1(p62) were detected by western blot.Results Compared with LPS group,PSAE could reduce the secretions of NO(P<0.05,P<0.01),decrease the mRNA expression level of iNOS,IL-1β,TNF-α,and MCP-1(P<0.05,P<0.01,P<0.001),and decrease the protein expression levels of iNOS(P<0.05,P<0.001),besides,PSAE could reduce the values of LC3Ⅱ/Ⅰ(P<0.01,P<0.001),and promote the protein expression of p62(P<0.01),thus reducing the autophagy levels.Conclusion PSAE could inhibit macrophage M1 polarization and autophagy levels of LPS-stimulated RAW264.7 cells,and alleviate cellular inflammation.

关 键 词：黄精水提物 巨噬细胞 极化 炎症 自噬 

分 类 号：R285[医药卫生—中药学]