脂肪因子WNT1诱导信号通路蛋白2对棕榈酸及脂多糖诱导的巨噬细胞极化的调节作用  

Regulation of palmitic acid and lipopolysaccharide-induced macrophage polarization by adipokine WNT1-inducible signaling pathway protein 2

作　　者：邓涯兰 毛敏 齐若梅[1] 赵卫 付紫晴 黎健[1] 陈北冬[1] Deng Yalan;Mao Min;Qi Ruomei;Zhao Wei;Fu Ziqing;Li Jian;Chen Beidong(The Key Laboratory of Geriatrics,Beijing Institute of Geriatrics,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing Hospital/National Center of Gerontology,National Health Commission,Beijing 100730,China;Beijing Union University,Beijing 100101,China)

机构地区：[1]北京医院、国家老年医学中心,国家卫生健康委北京老年医学研究所,国家卫生健康委老年医学重点实验室,中国医学科学院老年医学研究所,北京100730 [2]北京联合大学旅游学院,北京100101

出　　处：《中华老年医学杂志》2023年第5期563-569,共7页Chinese Journal of Geriatrics

基　　金：国家自然科学基金(82170890)。

摘　　要：目的探讨WNT1诱导信号通路蛋白2对棕榈酸以及脂多糖诱导的炎症中巨噬细胞极化的调节作用。方法用不同浓度的WISP2蛋白处理巨噬细胞系RAW264.7,采用Cell-Titer发光法测定细胞活力,确定WISP的作用浓度。将细胞分为对照组、棕榈酸处理组、棕榈酸联合不同浓度WISP2处理组(10μg/L及100μg/L)以及脂多糖处理组、脂多糖联合不同浓度WISP2处理组(10μg/L及100μg/L)。应用实时定量聚合酶链式反应检测各组巨噬细胞中M1及M2表型分子的mRNA水平;应用酶联免疫吸附实验检测巨噬细胞分泌的重要炎症因子肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6的蛋白水平。结果和对照组相比,10μg/L、100μg/L WISP2处理组均未对RAW264.7细胞的活性产生影响,却显著上调Tnfα(1.877±0.039、2.202±0.034,F=309.7,P<0.001)、Il6(1.418±0.056、1.506±0.059,F=81.39,P<0.001)、单核细胞趋化蛋白1(Mcp1)(1.620±0.014、1.982±0.125,F=71.45,P<0.001)、趋化因子c-c配体(Ccl3)(1.892±0.118、1.942±0.132,F=32.93,P<0.001)和诱导型-氧化氮合酶(iNos)(1.691±0.201、1.548±0.090,F=13.60,P<0.05)mRNA表达,显著下调抗炎因子Tgfβ(1.376±0.025、2.152±0.107,F=1.846,P<0.05)、CD206(2.123±0.031、3.139±1.663,F=8.037,P<0.05)、Il4(2.098±0.464、2.494±0.141,F=48.68,P<0.01)、Il10(1.303±0.216、1.574±0.274,F=5.774,P<0.05)mRNA表达,使其向M1型巨噬细胞极化;与对照组相比,100μmol/L棕榈酸能在转录及蛋白水平温和而显著地增加TNF-α、IL-6等炎症因子的表达;与棕榈酸单独刺激相比,给予WISP2联合处理进一步促进巨噬细胞炎症因子TNF-α[(589.4±17.0)ng/L、(692.6±83.4)ng/L,F=56.38,P<0.05]、IL-6[(15.13±1.14)ng/L、(13.33±1.22)ng/L,F=23.32,P<0.001]的蛋白表达及趋化因子Mcp1(160.0±9.8、140.0±18.9,F=141.1,P<0.0001)和Ccl3(17.76±1.92、14.41±1.27,F=125.2,P<0.0001)的mRNA的表达。与对照组相比,100μg/L脂多糖在蛋白水平强烈刺激巨噬细胞TNF-α[(3444±423)ng/L,F=71.20,P<0.0001]、IL-6[(49Objective To investigate the regulatory effect of WNT1-inducible signaling pathway protein 2(WISP2)on macrophage polarization in palmitic acid(PA)and lipopolysaccharide(LPS)-induced inflammation.Methods The macrophage cell line RAW264.7 was treated with different concentrations of WISP2 protein,and cell viability was determined by means of luminescence assay using Cell-Titer Glo to determine the concentration of WISP2.The cells were divided into control group,palmitic acid group,palmitic acid combined with different concentrations of WISP2 group(10μg/L and 100μg/L)and lipopolysaccharide group,lipopolysaccharide combined with different concentrations of WISP2 group(10μg/L and 100μg/L).mRNA expression of M1 and M2 macrophages phenotype of each group were detected by real-time quantitative polymerase chain reaction.The protein expression of important inflammatory factors,TNF-αand IL-6,were evaluated by ELISA.Results Compared with the control group,both 10μg/L and 100μg/L WISP2 groups had no effect on the activity of RAW264.7 cells,but significantly up-regulated the expression of various inflammatory factors,including Tnfα(1.877±0.039,2.202±0.034,F=309.7,P<0.001),Il6(1.418±0.056,1.506±0.059,F=81.39,P<0.001),Mcp1(1.620±0.014,1.982±0.125,F=71.45,P<0.001),Ccl3(1.892±0.118,1.942±0.132,F=32.93,P<0.001),and iNos(1.691±0.201,1.548±0.090,F=13.60,P<0.05).mRNA in macrophages,and significantly down-regulated the expression of anti-inflammatory factors,including Tgfβ(1.376±0.025,2.152±0.107,F=1.846,P<0.05),CD206(2.123±0.031,3.139±1.663,F=8.037,P<0.05),Il4(2.098±0.464,2.494±0.141,F=48.68,P<0.01),and Il10(1.303±0.216,1.574±0.274,F=5.774,P<0.05)mRNA,causing M1 type macrophage polarization.Compared with the control group,100μmol/L palmitic acid could mildly but significantly increase the expression of inflammatory factors such as TNF-αand IL-6 at the transcriptional and protein levels.Compared with palmitic acid stimulation alone,the combination of palmitic acid and WISP2 further promoted the protein exp

关 键 词：巨噬细胞极化 WNT1诱导信号通路蛋白2 棕榈酸 脂多糖 炎症反应 

分 类 号：R364.5[医药卫生—病理学]