CAJ | 학술논문

目的：探讨植物血凝素样氧化型低密度脂蛋白（ox-LDL）受体1（LOX-1）在肝窦内皮细胞（HSECs）中的表达和ox-LDL对其表达的调控作用。方法使用实时定量聚合酶链反应（RT-PCR）和Western blotting法从基因和蛋白水平检测未经处理HSECs 中LOX-1的表达。应用不同浓度ox-LDL（0、20、40、60、80和100 mg/L）对HSECs作用24 h并应用80 mg/L ox-LDL对HSECs作用不同时间（0、12、24和48 h），作用后实时定量PCR检测HSECs内LOX-1 mRNA的表达水平，Western blotting法检测细胞内LOX-1蛋白表达。给予80 mg/L ox-LDL干预组多聚肌苷酸250 mg/L作用24 h后，测定LOX-1 mRNA和蛋白的表达。采用单因素方差分析及t检验进行数据分析。结果 LOX-1 mRNA和蛋白在HSECs中均有表达。20～80 mg/L ox-LDL组HSECs中LOX-1 mRNA、蛋白表达水平随ox-LDL剂量增加而升高，与剂量有明显相关性（F＝38.7、3.43，均P＜0.05）。与80 mg/L ox-LDL组相比，100 mg/L ox-LDL 组 LOX-1 mRNA、蛋白表达下降，差异有统计学意义（ t ＝23.75、18.26， P ＜0.05）。80 mg/L ox-LDL对HSECs作用时间在0～24 h时，随着时间延长，LOX-1 mRNA、蛋白表达递增，与ox-LDL作用时间有明显相关性（F＝2.36、0.33，均P＜0.05）。与作用24 h相比，作用48 h组HSECs中LOX-1 mRNA、蛋白表达下降（t＝69.21、36.27，均P＜0.05）。与80 mg/L ox-LDL组相比，多聚肌苷酸组中LOX-1 mRNA和蛋白表达降低，两组差异均有统计学意义（ t＝54.93、28.19，均P＜0.05）。结论LOX-1存在于HSECs。在一定浓度和时间范围内，ox-LDL对HSECs LOX-1的调控作用具有时间和浓度依赖性，而多聚肌苷酸可部分抑制这种效应。
목적：탐토식물혈응소양양화형저밀도지단백（ox-LDL）수체1（LOX-1）재간두내피세포（HSECs）중적표체화ox-LDL대기표체적조공작용。방법사용실시정량취합매련반응（RT-PCR）화Western blotting법종기인화단백수평검측미경처리HSECs 중LOX-1적표체。응용불동농도ox-LDL（0、20、40、60、80화100 mg/L）대HSECs작용24 h병응용80 mg/L ox-LDL대HSECs작용불동시간（0、12、24화48 h），작용후실시정량PCR검측HSECs내LOX-1 mRNA적표체수평，Western blotting법검측세포내LOX-1단백표체。급여80 mg/L ox-LDL간예조다취기감산250 mg/L작용24 h후，측정LOX-1 mRNA화단백적표체。채용단인소방차분석급t검험진행수거분석。결과 LOX-1 mRNA화단백재HSECs중균유표체。20～80 mg/L ox-LDL조HSECs중LOX-1 mRNA、단백표체수평수ox-LDL제량증가이승고，여제량유명현상관성（F＝38.7、3.43，균P＜0.05）。여80 mg/L ox-LDL조상비，100 mg/L ox-LDL 조 LOX-1 mRNA、단백표체하강，차이유통계학의의（ t ＝23.75、18.26， P ＜0.05）。80 mg/L ox-LDL대HSECs작용시간재0～24 h시，수착시간연장，LOX-1 mRNA、단백표체체증，여ox-LDL작용시간유명현상관성（F＝2.36、0.33，균P＜0.05）。여작용24 h상비，작용48 h조HSECs중LOX-1 mRNA、단백표체하강（t＝69.21、36.27，균P＜0.05）。여80 mg/L ox-LDL조상비，다취기감산조중LOX-1 mRNA화단백표체강저，량조차이균유통계학의의（ t＝54.93、28.19，균P＜0.05）。결론LOX-1존재우HSECs。재일정농도화시간범위내，ox-LDL대HSECs LOX-1적조공작용구유시간화농도의뢰성，이다취기감산가부분억제저충효응。
Objective To investigate the presence of lectin-like oxidized low-density lipoprotein (ox-LDL) receptor-1 (LOX-1) in cultured hepatic sinusoidal endothelial cells (HSECs), and meanwhile discuss the regulation of its mechanisms under ox-LDL injury conditions.Methods The mRNA and protein expression of LOX-1 was testified by real-time quantitative polymerase chain reaction ( RT-PCR) and Western blotting methods.Meanwhile HSECs were stimulated with different concentration of ox-LDL (0, 20, 40, 60, 80 and 100 mg/L) for 24 h, and HSECs were stimulated with 80 mg/L for different times (0, 12, 24 and 48 h).Then expression of LOX-1 mRNA was measured by RT-PCR, and the protein expression of LOX-1 was testified by using Western blotting.Data were analyzed with one-way ANOVA and t test.Results LOX-1 exists in HSECs.After intervened with 20-80 mg/L ox-LDL for 24 h, LOX-1 mRNA and protein levels increased significantly in a dose-dependent manner (F=38.7, 3.48, both P <0.05).However, when concentration of ox-LDL reached 100 mg/L, the LOX-1 mRNA and protein expression decreased significantly compared with those intervened with 80 mg/L ox-LDL ( t =23.75,18.26, both P<0.05 ).Meanwhile, after intervened with 80 mg/L ox-LDL at different time (0-24 h), the expression of LOX-1 mRNA and protein significantly increased in a time-manner ( F =2.36, 0.328, both P <0.05 ).And compared with those in cells intervened for 24 h, the mRNA and protein expression of LOX-1 decreased significantly when intervened for 48 h(t=69.21, 36.27, both P<0.05).However, compared with the cells incubated with 80 mg/L ox-LDL, the mRNA and protein expression of LOX-1 significantly decreased in HSECs treated with polyinosinic cytidylic acid ( t=54.93, 28.19, both P<0.05 ).Conclusion LOX-1 exists in HSECs and ox-LDL could regulate it in a certain reach of concentration and time.However , polyinosinic cytidylic acid can partly restrained ox-LDL-induced LOX-1 expression.