CAJ | 학술논문

目的研究黄化倒水莲皂苷C对非埘称_二甲基精氨酸(ADMA)诱导的细胞间黏附因子(sICAM-1)表达的抑制作用及其机制. 方法应用酶联免疫吸附法测定内皮细胞培养液中的slCAM-1;应用凝胶迁移率实验法检测细胞内核因子кB(NF-кB)活性;采用荧光法检测细胞内活性氧水平. 结果 ADMA(30 μmol/L)孵育内皮细胞(12、24、48 h)呈时间依赖性地增加细胞上清液中sICAM-1[(138.02±16.40)、(194.52±11.14)、(274.28±13.11)ng/L]和活性氧含量[(75.64±5.22)、(100.18±11.15)、(107.23±13.45)U];ADMA(3～30μmol/L)孵育内皮细胞24 h呈浓度依赖性地增加细胞上清液中sICAM-1含量[(136.14±10.77)、(189.21±21.25)、(221.24μ23.45)ng/L];预先给予L-精氨酸0.5 mmol/L和吡咯烷二硫基甲酸盐10μmol/L可显著抑制上述效心;皂苷C(1、3、10 μmol/L)可呈浓度依赖性地抑制ADMA诱导的细胞内活性氧含量增加[(85.33±8.68)、(70.69±7.65)、(59.12±4.15)U]、NF-κB激活及sICAM-1[(336.58±23.32)、(203.27±25.18)、(174.13±14.53)ng/L]表达,预先给予L-精氨酸(0.5 mmol/L)和PDTC(10μmol/L)可抑制ADMA诱导的细胞内活性氧含量增加[(58.44±9.52)U和(54.12±4.47)U]、NF-κB激活及sICAM-1表达((132.56±12.96)ng/L和(136.17±11.18)ng/L]. 结论黄花倒水莲皂苷C可抑制ADMA所诱导的slCAM-1水平升高,该作用与抑制内皮细胞内活性氧/NF-κB途径有关.
목적 연구황화도수련조감C대비시칭_이갑기정안산(ADMA)유도적세포간점부인자(sICAM-1)표체적억제작용급기궤제. 방법응용매련면역흡부법측정내피세포배양액중적slCAM-1;응용응효천이솔실험법검측세포내핵인자кB(NF-кB)활성;채용형광법검측세포내활성양수평. 결과 ADMA(30 μmol/L)부육내피세포(12、24、48 h)정시간의뢰성지증가세포상청액중sICAM-1[(138.02±16.40)、(194.52±11.14)、(274.28±13.11)ng/L]화활성양함량[(75.64±5.22)、(100.18±11.15)、(107.23±13.45)U];ADMA(3～30μmol/L)부육내피세포24 h정농도의뢰성지증가세포상청액중sICAM-1함량[(136.14±10.77)、(189.21±21.25)、(221.24μ23.45)ng/L];예선급여L-정안산0.5 mmol/L화필각완이류기갑산염10μmol/L가현저억제상술효심;조감C(1、3、10 μmol/L)가정농도의뢰성지억제ADMA유도적세포내활성양함량증가[(85.33±8.68)、(70.69±7.65)、(59.12±4.15)U]、NF-κB격활급sICAM-1[(336.58±23.32)、(203.27±25.18)、(174.13±14.53)ng/L]표체,예선급여L-정안산(0.5 mmol/L)화PDTC(10μmol/L)가억제ADMA유도적세포내활성양함량증가[(58.44±9.52)U화(54.12±4.47)U]、NF-κB격활급sICAM-1표체((132.56±12.96)ng/L화(136.17±11.18)ng/L]. 결론황화도수련조감C가억제ADMA소유도적slCAM-1수평승고,해작용여억제내피세포내활성양/NF-κB도경유관.
Objective To investigate the inhibitory effect of reiniosidc C (RC) on asymmetric dimethylarginine (ADMA)-induced soluble interacellular adhesion molecule-1 (slCAM-1) expression and its mechanisms. Methods Human umbical vein endothelial cells (HUVEC 12) were cultured.The level of slCAM-1 in the conditioned medium was determined by ELISA. Changes in intracellular reactive oxygen species (ROS) levels were determined by measuring the oxidative conversion of cell permeable 2', 7'-dichlorofluorescein diacetate (DCFH-DA) to fluorescent dichlorofluorescein (DCF) in fluorospectro- photometer, and the nuclear factor-κB (NF-κB) DNA-binding activity was determined by electrophoretic mobility shift assays (EMSA). Results sICAM 1 expressions [(138.02±16.40), (194.52±11.14), (274.28±13.11)ng/L]and the generation of ROS[(75.64±5.22),(100.18±11.15),(107.23±13.45)units] in HUVEC-12 were time dependently increased by ADMA (30 μmol/L). Furthermore, thc generation of ROS [(85.33±8.68), (70.69±7.65),(59.12±4.15)units], activation of NF-κB activity and expression of sICAM-1 [(336.58±23.32),(203.27±25.18) ,(174.13±14.53)ng/L] induced by ADMA were inhibited by reinioside C (1,3,10μmol/L) in a dose-dependent manner. This effect was found to be the same by L-arginine (0.5 mmol/L) as NOS substrate and by pyrrolidine dithiocarbamate (PDTC) (10 μmol/L)as inhibitor of NF-κB.Conclusions Reinioside C attenuates the increase of sICAM-1 induced by exogenous ADMA