1-甲基色氨酸对脂多糖诱导的人脐静脉内皮细胞通透性及凋亡的影响  

Effect of 1-methyltryptophan on lipopolysaccharide-induced permeability and apoptosis in human umbilical vein endothelial cells

作　　者：王元元 许铎 沈继龙 周情太 赵慧静 陈亚利 汪明灯[1] Wang Yuanyuan;Xu Duo;Shen Jilong;Zhou Qingtai;Zhao Huijing;Chen Yali;Wang Mingdeng(Department of Intensive Care Unit,Suzhou Science&Technology Town Hospital Affiliated to NanjingMedical University,Suzhou 215153,China)

机构地区：[1]南京医科大学附属苏州科技城医院重症医学科,215153

出　　处：《中华急诊医学杂志》2021年第1期59-63,共5页Chinese Journal of Emergency Medicine

基　　金：苏州市产业技术创新专项(SYSD2017064)。

摘　　要：目的探讨1-甲基色氨酸(1-methyltryptophan,1-MT)对脂多糖(lipopolysaccharide,LPS)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)通透性及细胞凋亡的影响。方法将HUVECs分为三组,即磷酸缓冲盐溶液(phosphate buffered saline,PBS对照组)、1μg/mL LPS(LPS组)、含有LPS及1 mmol/L 1-MT(1-MT组)。采用蛋白免疫印迹试验(Western blot)检测孵育8 h后的p120连环(p120 concatemer,p120ctn)、血管内皮钙粘蛋白(vascular endothelial cadherin,VE-cadherin)、caspase-3及DNA修复酶多聚二磷酸腺苷核糖聚合酶-1(DNA repair enzyme polyadenylate ribose polymerase-1,PARP-1)的表达。采用高效液相色谱法(high performance liquid chromatography,HPLC)分别检测孵育2、4、6和8 h的犬尿氨酸(kynurenine,Kyn)浓度,并计算吲哚胺2,3-双加氧酶(indoleamine 2,3-dioxygenase,IDO)活性。组间两两均数比较用LSD-t检验。结果与PBS对照组相比较,LPS组caspase-3[(74.01±7.91)%vs(157.14±7.63)%,P<0.01]、Kyn表达显著增高,而p120ctn[(49.12±2.15)%vs(37.61±1.80)%,P<0.01]、VE-cadherin[(107.70±7.01)%vs(90.66±2.58)%,P=0.027]及抗凋亡蛋白PARP-1[(67.95±3.08)%vs(57.93±5.26)%,P=0.038]水平显著降低,IDO活性升高(P<0.05)。与LPS组比较,1-MT组caspase-3[(157.14±7.63)%vs(110.74±7.89)%,P<0.01]表达显著降低,然而p120ctn[(37.61±1.80)%vs(47.19±0.82)%,P<0.01]、VE-cadherin[(90.66±2.58)%vs(107.27±9.89)%,P=0.029]及PARP-1[(57.93±5.26)%vs(74.12±4.90)%,P=0.005]表达显著升高,不同时间点IDO活性均降低(P<0.05)。PBS组和1-MT组p120ctn、VE-cadherin、PARP-1蛋白水平、Kyn浓度及IDO活性差异无统计学意义(P>0.05),1-MT组caspase-3蛋白水平较PBS组升高(P=0.001)。结论LPS刺激HUVECs通透性增加,而1-MT可通过抑制IDO活性和降低Kyn水平拮抗LPS的刺激作用,此外,1-MT还可降低细胞凋亡,其机制可能是通过增加PARP-1表达,降低caspase-3水平,从而发挥内皮细胞的保护作用。Objective To explore the effect of 1-methyltryptophan(1-MT)on lipopolysaccharide(LPS)-induced permeability and apoptosis of human umbilical vein endothelial cells(HUVECs).Methods HUVECs were treated with phosphate buffer saline(PBS,control group),1μg/mL LPS(LPS group),and LPS combined with 1 mmol/L 1-MT(1-MT group).The expression levels of the p120 concatemer(p120ctn),vascular endothelial(VE)cadherin,caspase-3,and DNA repair enzyme polyadenylate ribose polymerase-1(PARP)after incubation at 8 h were detected using Western blot.The concentrations of kynurenine(Kyn)after incubation at 2,4,6,and 8 h were measured by highperformance liquid chromatography,and indoleamine2,3-dioxygenase(IDO)activity was calculated.Comparisons among groups were performed using the LSD-t test.Results Compared with the control group,the expression of caspase-3[(74.01±7.91)%vs(157.14±7.63)%,P<0.01]and the concentration of Kyn were significantly up-regulated,while the expression of p120ctn[(49.12±2.15)%vs(37.61±1.80)%,P<0.01],VE-cadherin[(107.70±7.01)%vs(90.66±2.58)%,P=0.027],and PARP-1[(67.95±3.08)%vs(57.93±5.26)%,P=0.038]were significantly down-regulated,and IDO activity was significantly increased in the LPS group(P<0.05).Compared with the LPS group,the expression of caspase-3[(157.14±7.63)%vs(110.74±7.89)%,P<0.01]was significantly down-regulated,while the expression of p120ctn[(37.61±1.80)%vs(47.19±0.82)%,P<0.01],VE-cadherin[(90.66±2.58)%vs(107.27±9.89)%,P=0.029],and PARP-1[(57.93±5.26)%vs(74.12±4.90)%,P=0.005]were significantly up-regulated,and the activity of IDO was significantly decreased over time in the 1-MT group(P<0.05).No significant differences were observed between the PBS and 1-MT groups in the protein levels of p120ctn,VEcadherin,and PARP-1 protein as well as Kyn concentration and IDO activity(P>0.05),while the expression of caspase-3 was increased in 1-MT group(P=0.001).Conclusions LPS aggravates the permeability of HUVECs,which can be reversed by 1-MT via inhibiting IDO activity and reducing Kyn concentrat

关 键 词：1-甲基色氨酸 脂多糖 脓毒症 内皮细胞 通透性 凋亡 

分 类 号：R459.7[医药卫生—急诊医学]