枸杞多糖对髓样分化因子88基因敲除小鼠2型糖尿病模型炎症因子的影响
|
摘要
目的·研究枸杞多糖(Lycium barbarum polysaccharide,LBP)对髓样分化因子88(myeloid differentiation factor 88,MyD88基因敲除(MyD88-/-)小鼠2型糖尿病(type 2 diabetes mellitus,T2DM)模型炎症因子的影响。方法·采用不同浓度LBP(20、40、80 mg/kg)干预MyD88-/-T2DM小鼠,ELISA法检测小鼠血清中白细胞介素1β(IL-1β)、IL-6、IL-8、转化生长因子β1(TGF-β1)及IL-10的水平。采用不同浓度(25、50、100μg/mL)LBP预处理小鼠巨噬细胞系Raw264.7细胞,然后用脂多糖诱导炎症状态,Western blotting检测各组细胞核因子κB(NF-κB)的核转位变化,以及NF-κB抑制蛋白(IκB)和磷酸化IκB的蛋白水平。结果·LBP能够降低MyD88-/-T2DM小鼠血清IL-1β和TGF-β1的水平(均P<0.05)。体外实验表明,LBP可以剂量依赖性地抑制巨噬细胞中由脂多糖诱发的NF-κB核转位,同时高剂量的LBP可以抑制IκB磷酸化。结论·LBP可以抑制MyD88-/-T2DM小鼠部分促炎症因子,这种调节作用可能与其弱化巨噬细胞IκB磷酸化,抑制NF-κB核转位有关。
Objective · To investigate the effect of Lycium barbarum polysaccharides on inflammatory cytokines in type 2 diabetes mellitus(T2 DM) mice without myeloid differentiation factor 88 gene(My D88-/-). Methods · Levels of interleukin 1β(IL-1β), IL-6, IL-8, transforming growth factor β1(TGF-β1),and IL-10 in serum were assessed by ELISA in the MyD88-/-T2 DM mice which had been administered with different doses of LBP(20, 40, and 80 mg/kg).Mouse macrophages Raw264.7 were stimulated by lipopolysaccharide(LPS) after treatment with different concentrations of LBP(25, 50, and 100 μg/mL).Then Western blotting was used to detect nuclear translocation level of nuclear factor κB(NF-κB) and protein expressions of inhibitor of NF-κB(IκB) and p-IκB. Results · Serum levels of IL-1β and TGF-β1 in MyD88-/-T2 DM mice were down-regulated by LBP(P<0.05). Cell experiment proved that nuclear migration of NF-κB was dose-dependently inhibited by LBP, and the level of p-IκB was reduced by high dose of LBP. Conclusion · LBP can reduce some proinflammatory cytokines in the MyD88-/-T2 DM mice, which may be related with its inhibitive effect on the phosphorylation of IκB and nuclear migration of NF-κB in the macrophages.
Objective · To investigate the effect of Lycium barbarum polysaccharides on inflammatory cytokines in type 2 diabetes mellitus(T2 DM) mice without myeloid differentiation factor 88 gene(My D88-/-). Methods · Levels of interleukin 1β(IL-1β), IL-6, IL-8, transforming growth factor β1(TGF-β1),and IL-10 in serum were assessed by ELISA in the MyD88-/-T2 DM mice which had been administered with different doses of LBP(20, 40, and 80 mg/kg).Mouse macrophages Raw264.7 were stimulated by lipopolysaccharide(LPS) after treatment with different concentrations of LBP(25, 50, and 100 μg/mL).Then Western blotting was used to detect nuclear translocation level of nuclear factor κB(NF-κB) and protein expressions of inhibitor of NF-κB(IκB) and p-IκB. Results · Serum levels of IL-1β and TGF-β1 in MyD88-/-T2 DM mice were down-regulated by LBP(P<0.05). Cell experiment proved that nuclear migration of NF-κB was dose-dependently inhibited by LBP, and the level of p-IκB was reduced by high dose of LBP. Conclusion · LBP can reduce some proinflammatory cytokines in the MyD88-/-T2 DM mice, which may be related with its inhibitive effect on the phosphorylation of IκB and nuclear migration of NF-κB in the macrophages.
引文
[1]Kontoangelos K,Papageorgiou CC,Paptis AE,et al.Cytokines,diabetes mellitus and psychopathology:a challenging combination[J].Neuro Endocrinol Lett,2014,35(2):159-169.
[2]Park YJ,Warnock GL,Ziliang AO,et al.Dual role of interleukin-1βin islet amyloid formation and itsβ-cell toxicity:implications for type 2 diabetes and islet transplantation[J].Diabetes Obes Metab,2017,19(5):682-694.
[3]Esser N,Paquot N,Scheen AJ.Anti-inflammatory agents to treat or prevent type 2 diabetes,metabolic syndrome and cardiovascular disease[J].Expert Opin Investig Drugs,2015,24(3):283-307.
[4]Giacomellia R,Ruscitti P,Alvaro S,et al.IL-1βat the crossroad between rheumatoid arthritis and type 2 diabetes:may we kill two birds with one stone?[J].Expert Rev Clin Immunol,2016,12(8):849-855.
[5]崔思远,李芳.IL-6与2型糖尿病关系的研究进展[J].中国微生态学杂志,2012,24(11):1051-1054.
[6]Kim D,Kang JM,Park SH,et al.Diabetes aggravates post-ischaemic renal fibrosis through persistent activation of TGF-β1 and Shh Signalling[J].Sci Rep,2017,7(1):16782.
[7]Wang Z,Shen X,Feng W,et al.Analysis of inflammatory mediators in prediabetes and newly diagnosed type 2 diabetes patients[J].J Diabetes Res,2016,2016:7965317.
[8]蔡慧珍,刘福康,芦慧霞,等.枸杞多糖对2型糖尿病病人的短期干预[J].食品科学,2012,33(13):259-262.
[9]张娜,李琳,高向晖,等.宁夏枸杞对代谢综合征患者TNF-α、NF-κB和i NOS水平的影响[J].中华疾病控制杂志,2014,18(4):286-289.
[10]Cai H,Liu F,Zuo P,et al.Practical application of antidiabetic efficacy of Lycium barbarum polysaccharide in patients with type 2 diabetes[J].Med Chem,2015,11(4):383-390.
[11]蔡慧珍,刘福康,孙桂菊.枸杞多糖对β-TC6细胞胰岛素分泌及相关基因的影响[J].江苏医药,2013,39(4):391-393.
[12]刘磊,李青旺,赵蕊,等.枸杞多糖对糖尿病小鼠胰岛细胞形态与功能的影响[J].黑龙江畜牧兽医,2008(3):93-94.
[13]余益本,王建平.TLR信号通路研究进展[J].细胞与分子免疫学杂志,2012,28(12):1331-1334.
[14]朱亚军.Toll样受体家族与高脂血症和胰岛素抵抗的相关性研究[D].石家庄:河北医科大学,2014.
[15]刘婷婷,王凌霄,杨晓辉,等.MyD88非依赖性信号通路在枸杞多糖抑制糖尿病小鼠肿瘤坏死因子α中的作用[J].浙江大学学报(医学版),2018,47(1):35-40.
[16]高春燕.枸杞多糖的提取分离技术及其特性研究[D].西安:陕西师范大学,2006.
[17]许金霞.枸杞多糖的制备及其活化巨噬细胞机理的研究[D].杭州:浙江大学,2007.
[18]刘源才,孙细珍,许银,等.枸杞多糖组成及含量测定方法的改进[J].食品科学,2013,34(12):292-295.
[19]向宇飞.糖尿病人群及高脂饮食肥胖小鼠的免疫代谢学研究[D].长沙:中南大学,2011.
[20]虞敏佳.MyD88在特定细胞型中对肥胖相关炎症疾病进展的作用[D].杭州:浙江大学,2013.
[21]Shen H,Wu N,Wang Y,et al.MyD88 gene knockout attenuates paraquatinduced acute lung injury[J].Toxicol Lett,2017,269:41-46.
[22]Bollyky PL,Bice JB,Sweet IR,et al.The Toll-like receptor signaling molecule Myd88 contributes to pancreaticβ-cell homeostasis in response to injury[J].PLoS One,2009,4(4):e5063.
[23]Borowska M,Dworacka M,Weso?owska A,et al.The impact of pharmacotherapy of type 2 diabetes mellitus on IL-1β,IL-6 and IL-10 secretion[J].Pharmacology,2016,97(3-4):189-194.
[24]秦凤,李春亮,张惠莉.二甲双胍对2型糖尿病模型大鼠胰岛素抵抗的影响[J].中国组织工程研究,2017(12):1909-1914.
[25]Rena G,Hardie DG,Pearson ER.The mechanisms of action of metformin[J].Diabetologia,2017,60(9):1577-1585.
[26]Sharma AK,Raikwar SK,Kurmi MK,et al.Gemfibrozil and its combination with metformin on pleiotropic effect on IL-10 and adiponectin and antiatherogenic treatment in insulin resistant type 2 diabetes mellitus rats[J].Inflammopharmacology,2013,21(2):137-145.
[27]Albert S,Baldwin J.The NF-κB and IκB proteins:new discoveries and insights[J].Annu Rev Immunol,1996,14:649-683.
[28]Oeckinghaus A,Hayden MS,Ghosh S.Crosstalk in NF-κB signaling pathways[J].Nat Immunol,2011,12(8):695-708.
[2]Park YJ,Warnock GL,Ziliang AO,et al.Dual role of interleukin-1βin islet amyloid formation and itsβ-cell toxicity:implications for type 2 diabetes and islet transplantation[J].Diabetes Obes Metab,2017,19(5):682-694.
[3]Esser N,Paquot N,Scheen AJ.Anti-inflammatory agents to treat or prevent type 2 diabetes,metabolic syndrome and cardiovascular disease[J].Expert Opin Investig Drugs,2015,24(3):283-307.
[4]Giacomellia R,Ruscitti P,Alvaro S,et al.IL-1βat the crossroad between rheumatoid arthritis and type 2 diabetes:may we kill two birds with one stone?[J].Expert Rev Clin Immunol,2016,12(8):849-855.
[5]崔思远,李芳.IL-6与2型糖尿病关系的研究进展[J].中国微生态学杂志,2012,24(11):1051-1054.
[6]Kim D,Kang JM,Park SH,et al.Diabetes aggravates post-ischaemic renal fibrosis through persistent activation of TGF-β1 and Shh Signalling[J].Sci Rep,2017,7(1):16782.
[7]Wang Z,Shen X,Feng W,et al.Analysis of inflammatory mediators in prediabetes and newly diagnosed type 2 diabetes patients[J].J Diabetes Res,2016,2016:7965317.
[8]蔡慧珍,刘福康,芦慧霞,等.枸杞多糖对2型糖尿病病人的短期干预[J].食品科学,2012,33(13):259-262.
[9]张娜,李琳,高向晖,等.宁夏枸杞对代谢综合征患者TNF-α、NF-κB和i NOS水平的影响[J].中华疾病控制杂志,2014,18(4):286-289.
[10]Cai H,Liu F,Zuo P,et al.Practical application of antidiabetic efficacy of Lycium barbarum polysaccharide in patients with type 2 diabetes[J].Med Chem,2015,11(4):383-390.
[11]蔡慧珍,刘福康,孙桂菊.枸杞多糖对β-TC6细胞胰岛素分泌及相关基因的影响[J].江苏医药,2013,39(4):391-393.
[12]刘磊,李青旺,赵蕊,等.枸杞多糖对糖尿病小鼠胰岛细胞形态与功能的影响[J].黑龙江畜牧兽医,2008(3):93-94.
[13]余益本,王建平.TLR信号通路研究进展[J].细胞与分子免疫学杂志,2012,28(12):1331-1334.
[14]朱亚军.Toll样受体家族与高脂血症和胰岛素抵抗的相关性研究[D].石家庄:河北医科大学,2014.
[15]刘婷婷,王凌霄,杨晓辉,等.MyD88非依赖性信号通路在枸杞多糖抑制糖尿病小鼠肿瘤坏死因子α中的作用[J].浙江大学学报(医学版),2018,47(1):35-40.
[16]高春燕.枸杞多糖的提取分离技术及其特性研究[D].西安:陕西师范大学,2006.
[17]许金霞.枸杞多糖的制备及其活化巨噬细胞机理的研究[D].杭州:浙江大学,2007.
[18]刘源才,孙细珍,许银,等.枸杞多糖组成及含量测定方法的改进[J].食品科学,2013,34(12):292-295.
[19]向宇飞.糖尿病人群及高脂饮食肥胖小鼠的免疫代谢学研究[D].长沙:中南大学,2011.
[20]虞敏佳.MyD88在特定细胞型中对肥胖相关炎症疾病进展的作用[D].杭州:浙江大学,2013.
[21]Shen H,Wu N,Wang Y,et al.MyD88 gene knockout attenuates paraquatinduced acute lung injury[J].Toxicol Lett,2017,269:41-46.
[22]Bollyky PL,Bice JB,Sweet IR,et al.The Toll-like receptor signaling molecule Myd88 contributes to pancreaticβ-cell homeostasis in response to injury[J].PLoS One,2009,4(4):e5063.
[23]Borowska M,Dworacka M,Weso?owska A,et al.The impact of pharmacotherapy of type 2 diabetes mellitus on IL-1β,IL-6 and IL-10 secretion[J].Pharmacology,2016,97(3-4):189-194.
[24]秦凤,李春亮,张惠莉.二甲双胍对2型糖尿病模型大鼠胰岛素抵抗的影响[J].中国组织工程研究,2017(12):1909-1914.
[25]Rena G,Hardie DG,Pearson ER.The mechanisms of action of metformin[J].Diabetologia,2017,60(9):1577-1585.
[26]Sharma AK,Raikwar SK,Kurmi MK,et al.Gemfibrozil and its combination with metformin on pleiotropic effect on IL-10 and adiponectin and antiatherogenic treatment in insulin resistant type 2 diabetes mellitus rats[J].Inflammopharmacology,2013,21(2):137-145.
[27]Albert S,Baldwin J.The NF-κB and IκB proteins:new discoveries and insights[J].Annu Rev Immunol,1996,14:649-683.
[28]Oeckinghaus A,Hayden MS,Ghosh S.Crosstalk in NF-κB signaling pathways[J].Nat Immunol,2011,12(8):695-708.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |