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 nucleotide-binding oligomerization domain-like receptor protein 3 and caspase-1 in human umbilical vein endothelial cells

摘要:

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1 μg/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。

Abstract:

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 (1 μg/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 concentration-dependent 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.