铁皮石斛糖蛋白对皮肤炎症期的调控作用及机制  

Regulatory Effect of Dendrobium officinale Kinura et Migo Glycoprotein on Skin Inflammation and Its Mechanism

作　　者：赵倩 戴天浥 洪文龙 周丽免 苏海冉 田洋[1,2,4] 白忠彬 ZHAO Qian;DAI Tian-yi;HONG Wen-long;ZHOU Li-mian;SU Hai-ran;TIAN Yang;BAI Zhong-bin(College of Food Science and Technology,Yunnan Agricultural University,Kunming 650201,China;National R&D Center for Moringa Processing Technology,Kunming 650201,China;Yunnan Huge Pharmaceutical Co.,Ltd.,Puer 665000,China;Food and Drug Resources Development and Utilization Engineering Center of the Ministry of Education,Kunming 650201,China)

机构地区：[1]云南农业大学食品科学技术学院,云南昆明650201 [2]云南农业大学国家辣木加工技术研发专业中心,云南昆明650201 [3]云南斛哥药业有限公司,云南普洱665000 [4]食药资源开发与利用教育部工程中心,云南昆明650201

出　　处：《食品工业科技》2020年第21期304-310,316,共8页Science and Technology of Food Industry

基　　金：云南农业大学人才引进科研启动项目(A2002320);云南省引进人才科研启动项目(A3007330);云南省中青年学术和技术带头人后备人才项目(2018HB040)。

摘　　要：研究铁皮石斛糖蛋白(DOKMG)对皮肤炎症期的调控作用及机制。本文以巨噬细胞株RAW264.7为研究对象,用铁皮石斛糖蛋白刺激巨噬细胞并做时间和浓度依赖性分析,应用实时荧光定量聚合酶链式反应(Real-time quantitative polymerase chain reaction,RT-PCR)检测炎症因子白介素-1β(Interleukin-1β,IL-1β)、白介素-6(Interleukin-6,IL-6)、肿瘤坏死因子(Tumor necrosis factor-α,TNF-α)以及炎症相关的环氧化酶-2(Cyclooxygenase-2,COX-2)和一氧化氮合酶(Nitric oxide synthase,iNOS)的mRNA表达水平。应用酶联免疫吸附反应(Enzyme-linked immunosorbent assay,ELISA)检测巨噬细胞培养上清中炎症因子的表达。采用蛋白印迹(Western blot,WB)分析铁皮石斛糖蛋白对炎症因子蛋白水平表达及核转录因子-κB(Nuclear transcription factor-κB,NF-κB)信号通路的影响。结果显示,铁皮石斛糖蛋白能在24 h后极显著促进IL-1,IL-6,TNF-α的基因表达(P<0.001),当浓度达到100μg/mL时促进最为明显,IL-1β相比对照组增加了10.09倍,IL-6增加了20.45倍,TNF-α增加了0.95倍。DOKMG对细胞上清中炎症因子TNF-α、IL-1β有促进作用,且成剂量依赖性,当浓度为100μg/mL时,两者分泌量分别由0.52μg/mL、118.52 pg/mL上升至1.99μg/mL、272.81 pg/mL。DOKMG能显著促进炎症因子蛋白的表达及NF-κB信号通路相关蛋白的磷酸化(P<0.05)。上述结果表明,铁皮石斛糖蛋白通过调节细胞炎症反应来促进伤口愈合,其部分机制与NF-κB信号通路激活有关。The aim of this study was to investigate the regulation and mechanism of Dendrobium officinale Kinura et Migo glycoprotein(DOKMG)on skin inflammation.In this paper,macrophage cell line RAW264.7 was targeted and stimulated by DOKMG in different time and concentration,the mRNA expression levels of inflammatory factor IL-1β,IL-6,TNF-αand inflammation-related COX-2 and iNOS were detected by RT-PCR.The expression of inflammatory factors in supernatant of macrophages was detected by ELISA.Western blot was used to analyze the effect of DOKMG on the expression of inflammatory factor protein and NF-κB signaling pathway.The results showed that DOKMG significantly promoted the expression of IL-1β,IL-6,TNF-αgenes after 24 h(P<0.001).When the concentration reached 100μg/mL,the improvement was the most obvious.Compared with the control group,IL-1βincreased by 10.09 times,IL-6 increased by 20.45 times,and TNF-αincreased by 0.95 times.When the concentration was 100μg/mL,the secretion of TNF-α、IL-1βincreased from 0.52μg/mL and 118.52 pg/mL to 1.99μg/mL and 272.81 pg/mL,the expression of inflammatory cytokines and phosphorylation of NF-κB were significantly promoted(P<0.05).These results indicate that DOKM can significantly promote macrophage inflammatory response to promote wound healing,and its mechanism may be related to the activation of NF-κB signaling pathway.

关 键 词：铁皮石斛 糖蛋白 炎症因子 核转录因子(NF-κB) 伤口愈合 

分 类 号：TS201.4[轻工技术与工程—食品科学]