沙葱总黄酮对脂多糖诱导的小鼠腹腔巨噬细胞炎症介质的影响

doi:10.3969/j.issn.1006-267x.2018.09.041

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摘要本试验以脂多糖（LPS）诱导的小鼠腹腔巨噬细胞为炎症模型，旨在研究沙葱总黄酮的抗炎作用。应用CCK-8法筛选沙葱总黄酮对小鼠腹腔巨噬细胞活力具有促进作用的浓度，在此基础上设置对照组、LPS组（应激模型，1 μg/mL LPS）、沙葱总黄酮低剂量组（25 μg/mL沙葱总黄酮+1 μg/mL LPS）、沙葱总黄酮中剂量组（50 μg/mL沙葱总黄酮+1 μg/mL LPS）、沙葱总黄酮高剂量组（100 μg/mL沙葱总黄酮+1 μg/mL LPS）。用Griess法测定细胞上清液中一氧化氮（NO）含量；用酶联免疫吸附试验（ELISA）法测定细胞上清液中肿瘤坏死因子-α（TNF-α）、白介素（IL）-6、IL-1β、IL-10含量；用逆转录聚合酶链式反应（RT-PCR）法测定细胞中TNF-α、IL-6、IL-1β、IL-10、一氧化氮合酶（iNOS）mRNA表达量。结果表明：1）与对照组相比，沙葱总黄酮的添加浓度在25.0~100.0 μg/mL时均能显著提高细胞增殖率（P<0.05）；2）与对照组相比，LPS组能显著提高细胞上清液NO的含量（P<0.05）；与LPS组相比，不同浓度沙葱总黄酮均能显著抑制NO的产生（P<0.05）；3）与对照组相比，LPS组的细胞上清液中TNF-α、IL-6、IL-1β、IL-10含量以及细胞中TNF-α、IL-6、IL-1β、IL-10、iNOS mRNA表达量均显著提高（P<0.05）；与LPS组相比，除了低剂量组IL-1β外，25、50、100 μg/mL沙葱总黄酮浓度显著降低细胞上清液中TNF-α、IL-6、IL-1β含量（P<0.05），除了低剂量组IL-10外，显著增加IL-10含量（P<0.05）；除了低剂量组iNOS外，25、50、100 μg/mL沙葱总黄酮浓度显著抑制细胞中TNF-α、IL-6、IL-1β、iNOS mRNA的表达（P<0.05），有促进IL-10 mRNA表达的趋势，但差异不显著（P>0.05）。综上，沙葱总黄酮对LPS诱导的小鼠腹腔巨噬细胞具有显著的抗炎作用。

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关键词 ：沙葱, 总黄酮, 脂多糖, 小鼠, 腹腔巨噬细胞, 应激模型

Abstract：In the present study, mouse peritoneal macrophages (MPM) induced by lipopolysaccharide (LPS) were used as an inflammation model to investigate the anti-inflammatory effect of total flavonoids from Allium mongolicum Regel. The CCK-8 method was used to select the concentrations of total flavonoids that could stimulate cell viability of MPM. Based on the concentrations, the cells were treated in control group, LPS group (stress model, 1 μg/mL LPS), total flavonoids low dosage group (25 μg/mL total flavonoids+1 μg/mL LPS), medium dosage group (50 μg/mL total flavonoids+1 μg/mL LPS) and high dosage group (100 μg/mL total flavonoids+1 μg/mL LPS). The Griess method was applied to examine the nitric oxide (NO) content in cell supernatant; the contents of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1β and IL-10 in cell supernatant were examined by the method of ELISA; the mRNA expression levels of TNF-α, IL-6, IL-1β, IL-10 and nitric oxide synthase (iNOS) of cells were tested by reverse transcription and polymerase chain reaction (RT-PCR). The results showed as follows:1) compared with the control, the cell proliferation ratio was significantly increased when the concentration range of total flavonoids was 25.0 to 200.0 μg/mL (P<0.05). 2) Compared with the control, the content of NO in cell supernatant was significantly increased in LPS group (P<0.05); compared with LPS group, different concentrations of total flavonoids could inhibit NO production (P<0.05). 3) Compared with the control, the contents of TNF-α,IL-6,IL-1β,IL-10 in cell supernatant and mRNA expression levels of TNF-α, IL-6, IL-1β, IL-10, iNOS in cells were significantly increased in LPS group (P<0.05); compared with LPS group, when cells were treated with total flavonoids at concoctions of 25, 50 and 100 μg/mL, except for IL-1β in low dosage group, the contents of TNF-α, IL-6 and IL-1β in cell supernatant were significantly decreased (P<0.05), and IL-10 content was significantly increased (P<0.05); except for iNOS in high dosage group, total flavonoids at concoctions of 25, 50 and 100 μg/mL significantly inhibited the mRNA expression of TNF-α,IL-6,IL-1β,iNOS in cells (P<0.05), and trend to improve IL-10 mRNA expression, but the difference was not significant (P>0.05). In conclusion, total flavonoids from Allium mongolicum Regel have a significant anti-inflammatory effect on MPM induced by LPS.

Key words： Allium mongolicum Regel total flavonoids LPS mouse peritoneal macrophages stress model

收稿日期: 2018-03-12

PACS:

S816

基金资助:

国家自然科学基金项目（31260558）

通讯作者: 敖长金,教授,博士生导师,E-mail:changjinao@aliyun.com E-mail: changjinao@aliyun.com

作者简介: 王特日格乐(1994-),男,内蒙古通辽人,硕士研究生,从事动物营养与饲料研究。E-mail:843401225@qq.com

引用本文:

王特日格乐, 王翠芳, 丹妮, 萨茹丽, 杜红喜, 郭春利, 哈斯额尔敦, 曹琪娜, 敖长金. 沙葱总黄酮对脂多糖诱导的小鼠腹腔巨噬细胞炎症介质的影响[J]. 动物营养学报, 2018, 30(9): 3702-3709.
WANG Terigele, WANG Cuifang, DAN Ni, SA Ruli, DU Hongxi, GUO Chunli, Khas-Erdene, CAO Qina, AO Changjin. Effects of Total Flavonoids from Allium mongolicum Regel on Inflammatory Mediators induced by Lipopolysaccharide of Mouse Peritoneal Macrophages. Chinese Journal of Animal Nutrition, 2018, 30(9): 3702-3709.

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http://www.chinajan.com/CN/10.3969/j.issn.1006-267x.2018.09.041 或 http://www.chinajan.com/CN/Y2018/V30/I9/3702

[1]	林曦.家畜病理学[M].3版.北京:中国农业出版社,1997:74-99.
[2]	代艳文,袁丁,万静枝,等.竹节参总皂苷通过NF-κB通路对LPS致RAW264.7细胞炎症的保护作用研究[J].中国中药杂志,2014,39(11):2076-2080.
[5]	中国科学院中国植物志编辑委员会.中国植物志(14卷)[M].北京:科学出版社,1980:170-172.
[6]	巴俊杰,张春丽,高建萍,等.沙葱营养成分分析[J].内蒙古农业大学学报(自然科学版),2002,23(4):114-115.
[7]	木其尔.沙葱黄酮对肉羊机体抗氧化和免疫功能的影响及其机理研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2016.
[8]	鄢春旻.黄酮类化合物抗病毒及抗炎活性研究[D].硕士学位论文.南京:南京大学,2012.
[10]	萨茹丽.沙葱黄酮提取工艺优化、结构鉴定及其相关生物活性研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2014.
[11]	刘博.小鼠巨噬细胞TLR2、TLR4及RP105在金黄色葡萄球菌感染中的天然免疫应答机制[D].博士学位论文.长春:吉林大学,2013.
[25]	ADEREM A,UNDERHILL D M.Mechanisms of phagocytosis in macrophages[J].Annual Review of Immunology,17(1):593-623.
[28]	杨晓露,刘朵,卞卡,等.甘草总黄酮及其成分体外抗炎活性及机制研究[J].中国中药杂志,2013,38(1):99-104.
[29]	房蓓蓓.柑桔皮渣中黄酮类化合物的提取及其抗炎活性研究[D].硕士学位论文.重庆:重庆工商大学,2008.
[32]	杨彩霞.茼麻叶总黄酮体外抗炎活性的研究[D].硕士学位论文.沈阳:沈阳农业大学,2016.
[15]	徐艺荣.九种黄酮类化合物对LPS诱导的RAW264.7细胞PGE2、COX-2表达的影响[D].硕士学位论文.天津:天津科技大学,2012.
[16]	DONG D,ZHOU N N,PAN H X,et al.Sarsasapogenin-AA13 inhibits LPS-induced inflammatory responses in macrophage cells in vitro and relieves dimethylbenzene-induced ear edema in mice[J].Acta Pharmacologica Sinica,2017,38(5):699-709.
[24]	MARTIN P,LEIBOVICH S J.Inflammatory cells during wound repair:the good,the bad and the ugly[J].Trendsin in Cell Biology,2005,15(11):599-607.
[27]	MARSH C B,WEWERS M D.The pathogenesis of sepsis:factors that modulate the response to gram-negative bacterial infection[J].Clinics in Chest Medicine,1996,17(2):183-197.
[33]	林泽华.侧柏叶黄酮和多糖分离纯化、结构表征及活性评价[D].硕士学位论文.广州:华南理工大学,2016.
[34]	杨杰,沙金丹,高翔,等.黄酮类化合物的免疫调节作用及机制[J].动物营养学报,2017,29(12):4295-4300.
[4]	程亮.褪黑素诱导肝癌细胞释放的外泌体对巨噬细胞免疫功能的影响及机制研究[D].硕士学位论文.合肥:安徽医科大学,2017.
[9]	王月乔.新型黄酮衍生物的合成研究[D].硕士学位论文.郑州:河南大学,2015.
[13]	FAN G W,JIANG X R,WU X Y,et al.Anti-inflammatory activity of tanshinone ⅡA in LPS-stimulated RAW264.7 macrophages via miRNAs and TLR4-NF-κB pathway[J].Inflammation,2016,39(1):375-384.
[14]	武剑.柴胡多糖对巨噬细胞免疫功能的调节及对TLR4信号通路的影响[D].硕士学位论文.上海:复旦大学,2012.
[19]	原俊.丹酚酸A通过抑制NF-κB的活化在小鼠腹腔巨噬细胞中的抗炎作用[D].硕士学位论文.大连:大连医科大学,2008.
[20]	BABCOCK A A,KUZIEL W A,RIVEST S O,et al.Chemokine expression by glial cells directs leukocytes to sites of axonal injury in the CNS[J].Journal of Neuroscience,2003,23(21):7922-7930.
[21]	SIMS J E,SMITH D E.The IL-1 family:regulators of immunity[J].Nature Reviews Immunology,2010,10(2):89-102.
[31]	郭轶.血栓闭塞性脉管炎发病机制的研究进展[J].重庆医科大学学报,2007,32:176-178.
[3]	VODOVOTZ Y,CONSTANTINE G,FAEDER J,et al.Translational systems approaches to the biology of inflammation and healing[J].Immunopharmacology and Immunotoxicology,2010,32(2):181-195.
[12]	RAY A,DITTEL B N.Isolation of mouse peritoneal cavity cells[J].Journal of Visualized Experiments,2010(35):1488.
[17]	KIM J S,JOBIN C.The flavonoid luteolin prevents lipopolysaccharide-induced NF-κB signaling and gene expression by blocking IκB kinase activity in intestinal epithelial cells and bone-marrow derived dendritic cell[J].Immunology,2005,115(3):357-387.
[18]	CHEN X J,YANG X F,LIU T J,et al.Kaempferol regulates MAPKs and NF-κB signaling pathways to attenuate LPS-induced acute lung injury in mice[J].International Immunopharmacology,2012,14(2):209-216.
[22]	YEE L J,TANG J M,GIBSON A W,et al.Interleukin 10 polymorphisms as predictors of sustained response in antiviral therapy for chronic hepatitis C infection[J].Hepatology,2001,33(3):708-712.
[23]	NORMAN J.The role of cytokines in the pathogenesis of acute pancreatitis[J].The American Journal of Surgery,1998,175(1):76-83.
[26]	SHAPIRO H,LUTATY A,ARIEL A.Macrophages,meta-inflammation,and immuno-metabolism[J].The Scientific World Journal,2011,11:2509-2529.
[30]	MURRAY P J,WYNN T A.Protective and pathogenic functions of macrophage subsets[J].Nature Reviews Immunology,2011,11(11):723-737.