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

Effect of berberine on lipopolysaccharide-induced inflammatory response in human umbilical vein endothelia cells

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(250,125,62.5,31.25,15.625,7.8125,3.90625,1.953125 and 0.9765μmol/L) of berberine,the proliferation of HUVECs was detected by methyl thiazolyl tetrazolium(MTT)assay.HUVECs were divided into blank group,lipopolysaccharide group,lipopolysaccharide+1.25μmol/L berberine group,lipopolysaccharide+2.50μmol/L berberine group and lipopolysaccharide+5.00μmol/L berberine group.The blank group received the equivalent volume of phosphate buffered saline,other4groups were treated with corresponding drugs in which the concentration of lipopolysaccharide for intervention was defined as0.05μg/ml.The levels of interleukin(IL)-6and the expression levels of Toll-like receptor4(TLR4),cysteinyl aspartate specific proteinase(Caspase)-11,Caspase-1,NOD-like receptor3(NLRP3),apoptosis-associated speck-like protein containing CARD(ASC),high-mobility group box-1(HMGB1)and IL-1βproteins 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)of5μmol/L approximately.The levels of IL-6of the lipopolysaccharide group,the lipopolysaccharide+5.00μmol/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,HMGB1and IL-1β(all P<0.05),excluding NLRP3-ASC in the lipopolysaccharide+1.25μmol/L berberine group.Conclusion Berberine alleviates lipopolysaccharide-induced inflammation response of HUVECs probably by inhibiting Caspase-11-mediated atypical NLRP3inflammasome pathway.