降钙素原对脂多糖诱导的人脐静脉内皮细胞NLRP3和caspase-1表达的影响

Effect of procalcitonin on lipopolysaccharide-induced expression of nucleotidebinding oligomerization domain-like receptor protein 3 and caspase-1 in human umbilical vein endothelial cells

目的 探讨降钙素原(procalcitonin,PCT)对脂多糖(lipopolysaccharide,LPS)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)焦亡相关蛋白核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)、半胱氨酸天冬氨酸蛋白酶-1 (caspase-1)表达的影响。方法 以LPS诱导HUVECs建立脓毒症内皮细胞炎症损伤模型。实验分为两部分:(1)将HUVECs随机分成正常对照组、LPS组(浓度1μg/mL)、PCT组(浓度10 ng/mL)及LPS+PCT组(各组n=3);(2)正常对照组、LPS组、LPS+PCT不同浓度(0.1、1、10、100 ng/mL)组(各组n=3)。采用实时荧光定量聚合酶链反应法和Western blot法检测各组细胞NLRP3、caspase-1 mRNA及其蛋白的表达。结果 (1)与正常对照组比较,LPS组、PCT组及LPS+PCT组NLRP3、caspase-1 mRNA和蛋白表达均上调(P<0.05);与LPS组比较,LPS+PCT组NLRP3、caspase-1 mRNA和蛋白表达均下调(P<0.05)。(2)与LPS组比较,LPS+PCT不同浓度组NLRP3、caspase-1 mRNA和蛋白表达下调;随PCT浓度增加,NLRP3、caspase-1 mRNA及蛋白表达逐渐下调(P<0.05)。结论 LPS可促进HUVECs焦亡相关因子NLRP3、caspase-1表达,PCT干预可抑制LPS诱导的HUVECs中焦亡相关蛋白NLRP3、caspase-1的表达,并呈浓度依赖性。

Objective To study the effect of procalcitonin(PCT)on lipopolysaccharide(LPS)-induced expression of the pyroptosis-related proteins nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)and caspase-1 in human umbilical vein endothelial cells(HUVECs).Methods HUVECs were induced by LPS to establish a model of sepsis-induced inflammatory endothelial cell injury.The experiment was divided into two parts.In the first part,HUVECs were randomly divided into four groups:normal control,LPS(1μg/mL),PCT(10 ng/mL),and LPS+PCT(n=3 each).In the second part,HUVECs were randomly grouped:normal control,LPS,and LPS+PCT of different concentrations(0.1,1,10,and 100 ng/mL)(n=3 each).Quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of NLRP3 and caspase-1 in each group.Results In the first experiment:compared with the normal control group,the PCT,LPS,and LPS+PCT groups had significantly upregulated mRNA and protein expression levels of NLRP3 and caspase-1(P<0.05);compared with the LPS group,the LPS+PCT group had significantly downregulated mRNA and protein expression levels of NLRP3 and caspase-1(P<0.05).In the second experiment:compared with those in the LPS group,the mRNA and protein expression levels of NLRP3 and caspase-1 in the LPS+PCT of different concentrations groups were significantly downregulated in a concentrationdependent manner(P<0.05).Conclusions LPS can promote the expression of the pyroptosis-related proteins NLRP3 and caspase-1 in HUVECs,while PCT can inhibit the LPS-induced expression of the pyroptosis-related proteins NLRP3 and caspase-1 in HUVECs in a concentration-dependent manner.