小檗碱对脂多糖诱导的人脐静脉血管内皮细胞炎症反应的影响
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摘要
目的探讨小檗碱对脂多糖诱导的人脐静脉血管内皮细胞(HUVECs)炎症反应的影响。方法应用不同浓度(250、125、62. 5、31. 25、15. 625、7. 812 5、3. 906 25、1. 953 125、0. 976 5μmol/L)的小檗碱干预HUVECs后,采用四甲基偶氮唑盐法检测HUVECs的增殖情况。将HUVECs分为空白组、脂多糖组、脂多糖+1. 25μmol/L小檗碱组、脂多糖+2. 50μmol/L小檗碱组、脂多糖+5. 00μmol/L小檗碱组。空白组加入等体积磷酸缓冲液,其余4组加入相应药物干预,其中脂多糖的干预浓度为0. 05μg/ml。比较5组白细胞介素(IL)-6水平及Toll样受体4(TLR4)、半胱氨酸天冬氨酸蛋白酶(Caspase)-11、Caspase-1、NOD样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、高迁移率族蛋白1 (HMGB1)、IL-1β蛋白表达水平。结果不同浓度小檗碱对HUVECs增殖均具有抑制作用,半抑制浓度接近5. 00μmol/L。脂多糖组、脂多糖+5. 00μmol/L小檗碱组、空白组IL-6水平依次降低(P <0. 01)。除脂多糖+1. 25μmol/L小檗碱组中NLRP3-ASC外,其余脂多糖+不同浓度小檗碱组TLR4、Caspase-11、Caspase-1、NLRP3、ASC、HMGB1和IL-1β的表达水平均较脂多糖组降低(均P <0. 05)。结论小檗碱可能通过抑制Caspase-11介导的非经典的NLRP3炎症小体通路来减轻脂多糖诱导的HUVECs炎症反应。
Objective To explore the effect of berberine on lipopolysaccharide-induced inflammatory response in human umbilical vein endothelia cells( HUVECs). Methods After HUVECs intervention by different concentrations( 250,125,62. 5,31. 25,15. 625,7. 812 5,3. 906 25,1. 953 125 and 0. 976 5 μmol/L) of berberine,the proliferation of HUVECs was detected by methyl thiazolyl tetrazolium( MTT) assay. HUVECs were divided into blank group,lipopolysaccharide group,lipopolysaccharide + 1. 25 μmol/L berberine group,lipopolysaccharide + 2. 50 μmol/L berberine group and lipopolysaccharide + 5. 00 μmol/L berberine group. The blank group received the equivalent volume of phosphate buffered saline,other 4 groups were treated with corresponding drugs in which the concentration of lipopolysaccharide for intervention was defined as 0. 05 μg/ml. The levels of interleukin( IL)-6 and the expression levels of Toll-like receptor4( TLR4),cysteinyl aspartate specific proteinase( Caspase)-11,Caspase-1,NOD-like receptor 3( NLRP3),apoptosis-associated speck-like protein containing CARD( ASC),high-mobility group box-1( HMGB1) and IL-1β proteins were compared among the five groups.Results Different concentrations of berberine had an inhibitory effect on the proliferation of HUVECs,with the half maximal inhibitory concentration( IC50) of 5 μmol/L approximately. The levels of IL-6 of the lipopolysaccharide group,the lipopolysaccharide + 5. 00 μmol/L berberine group and the blank group deceased in turn( P < 0. 05). Compared with the lipopolysaccharide group,the lipopolysaccharide +berberine groups of different concentrations had lower expression levels of TLR4,Caspase-11,Caspase-1,NLRP3,ASC,HMGB1 and IL-1β( all P < 0. 05),excluding NLRP3-ASC in the lipopolysaccharide + 1. 25 μmol/L berberine group. Conclusion Berberine alleviates lipopolysaccharide-induced inflammation response of HUVECs probably by inhibiting Caspase-11-mediated atypical NLRP3 inflammasome pathway.
Objective To explore the effect of berberine on lipopolysaccharide-induced inflammatory response in human umbilical vein endothelia cells( HUVECs). Methods After HUVECs intervention by different concentrations( 250,125,62. 5,31. 25,15. 625,7. 812 5,3. 906 25,1. 953 125 and 0. 976 5 μmol/L) of berberine,the proliferation of HUVECs was detected by methyl thiazolyl tetrazolium( MTT) assay. HUVECs were divided into blank group,lipopolysaccharide group,lipopolysaccharide + 1. 25 μmol/L berberine group,lipopolysaccharide + 2. 50 μmol/L berberine group and lipopolysaccharide + 5. 00 μmol/L berberine group. The blank group received the equivalent volume of phosphate buffered saline,other 4 groups were treated with corresponding drugs in which the concentration of lipopolysaccharide for intervention was defined as 0. 05 μg/ml. The levels of interleukin( IL)-6 and the expression levels of Toll-like receptor4( TLR4),cysteinyl aspartate specific proteinase( Caspase)-11,Caspase-1,NOD-like receptor 3( NLRP3),apoptosis-associated speck-like protein containing CARD( ASC),high-mobility group box-1( HMGB1) and IL-1β proteins were compared among the five groups.Results Different concentrations of berberine had an inhibitory effect on the proliferation of HUVECs,with the half maximal inhibitory concentration( IC50) of 5 μmol/L approximately. The levels of IL-6 of the lipopolysaccharide group,the lipopolysaccharide + 5. 00 μmol/L berberine group and the blank group deceased in turn( P < 0. 05). Compared with the lipopolysaccharide group,the lipopolysaccharide +berberine groups of different concentrations had lower expression levels of TLR4,Caspase-11,Caspase-1,NLRP3,ASC,HMGB1 and IL-1β( all P < 0. 05),excluding NLRP3-ASC in the lipopolysaccharide + 1. 25 μmol/L berberine group. Conclusion Berberine alleviates lipopolysaccharide-induced inflammation response of HUVECs probably by inhibiting Caspase-11-mediated atypical NLRP3 inflammasome pathway.
引文
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[21] Zhang H,Shan Y,Wu Y,et al. Berberine suppresses LPSinduced inflammation through modulating Sirt1/NF-κB signaling pathway in RAW264. 7 cells[J]. Int Immunopharmacol,2017,52:93-100.
[22]张晨光,任峰,王辉. HMGB1与其受体分子信号转导通路的研究进展[J].中国免疫学杂志,2014,30(2):275-279.
[2]戴谆,宋金春,杨小青.甲基莲心碱对脂多糖诱导的人脐静脉内皮细胞的保护作用[J].中国药师,2016,19(10):1 875-1 878.
[3]侯宏,孙胜亮,黄静,等.黄连生物碱抗高脂血症及动脉粥样硬化实验研究[J].时珍国医国药,2011,22(10):2 462-2 464.
[4]万强,周凤华,贾钰华,等.小檗碱抑制p38 MAPK通路降低visfatin诱导HUVEC损伤的研究[J].中药新药与临床药理,2015,26(5):586-590.
[5]牟娜娜,娄晓盈,王仁庆,等. LPS激活巨噬细胞NLRP3炎性小体的作用研究[J].热带医学杂志,2015,15(4):441-443,466.
[6] Strowig T,Henao-Mejia J,Elinav E,et al. Inflammasomes in health and disease[J]. Nature,2012,481(7 381):278-286.
[7] Próchnicki T,Latz E. Inflammasomes on the crossroads of innate immune recognition and metabolic control[J]. Cell Metab,2017,26(1):71-93.
[8] Broz P. Inflammasomes in host defense and autoimmunity[J]. Chimia(Aarau),2016,70(12):853-855.
[9] Zhu Y,Zhu H,Wang Z,et al. Wogonoside alleviates inflammation induced by traumatic spinal cord injury by suppressing NF-κB and NLRP3 inflammasome activation[J]. Exp Ther Med,2017,14(4):3 304-3 308.
[10] Song N,Liu ZS,Xue W,et al. NLRP3 phosphorylation is an essential priming event for inflammasome activation[J]. Mol Cell,2017,68(1):185-197.
[11] Subramanian N,Natarajan K,Clatworthy MR,et al. The adaptor MAVS promotes NLRP3 mitochondrial localization and inflammasome activation[J]. Cell,2013,153(2):348-361.
[12] Pellegrini C,Antonioli L,Lopez-Castejon G,et al. Canonical and non-canonical activation of NLRP3 inflammasome at the crossroad between immune tolerance and intestinal inflammation[J]. Front Immunol,2017,8:36.
[13] Wang H,Xu YS,Wang ML,et al. Protective effect of naringin against the LPS-induced apoptosis of PC12 cells:implications for the treatment of neurodegenerative disorders[J].Int J Mol Med,2017,39(4):819-830.
[14] Jo EK,Kim JK,Shin DM,et al. Molecular mechanisms regulating NLRP3 inflammasome activation[J]. Cell Mol Immunol,2016,13(2):148-159.
[15] Hughes MM,O'neill LAJ. Metabolic regulation of NLRP3[J]. Immunol Rev,2018,281(1):88-98.
[16] Man SM,Karki R,Sasai M,et al. IRGB10 liberates bacterial ligands for sensing by the AIM2 and caspase-11-NLRP3 inflammasomes[J]. Cell,2016,167(2):382-396.
[17] Song E,Jahng JW,Chong LP,et al. Lipocalin-2 induces NLRP3 inflammasome activation via HMGB1 induced TLR4 signaling in heart tissue of mice under pressure overload challenge[J]. Am J Transl Res,2017,9(6):2 723-2 735.
[18] Rühl S,Broz P. Caspase-11 activates a canonical NLRP3 inflammasome by promoting K+efflux[J]. Eur J Immunol,2015,45(10):2 927-2 936.
[19]张红霞,赵娟,于希忠,等.小檗碱和人参皂苷Rb_1合用对脂肪细胞炎症因子表达和炎症信号通路的影响[J].食品与生物技术学报,2014,33(6):597-603.
[20]刘司漩.小檗碱对脂多糖诱导的THP-I细胞相关炎症因子的影响[D].太原:山西医科大学,2014.
[21] Zhang H,Shan Y,Wu Y,et al. Berberine suppresses LPSinduced inflammation through modulating Sirt1/NF-κB signaling pathway in RAW264. 7 cells[J]. Int Immunopharmacol,2017,52:93-100.
[22]张晨光,任峰,王辉. HMGB1与其受体分子信号转导通路的研究进展[J].中国免疫学杂志,2014,30(2):275-279.