中国临床药理学与治疗学
中國臨床藥理學與治療學
중국림상약이학여치료학
CHINESE JOURNAL OF CLINICAL PHARMACOLOGY
2008年
6期
663-670
,共8页
西红花酸%晚期糖基化终产物(AGE)%晚期糖基化终产物受体(RAGE)%活性氧(ROS)%内皮细胞%细胞间黏附分子-1(ICAM-1)
西紅花痠%晚期糖基化終產物(AGE)%晚期糖基化終產物受體(RAGE)%活性氧(ROS)%內皮細胞%細胞間黏附分子-1(ICAM-1)
서홍화산%만기당기화종산물(AGE)%만기당기화종산물수체(RAGE)%활성양(ROS)%내피세포%세포간점부분자-1(ICAM-1)
crocetin%advanced glycation end products(AGE)%receptor for AGE(RAGE)%reactive oxygen species(ROS)%endothelial cells%ICAM-1 protein
目的:研究西红花酸对晚期糖基化终产物(AGE)诱导牛内皮细胞(BEC)中晚期糖基化终产物受体(RAGE)mRNA表达的抑制作用,并探讨其可能机制.方法:不同浓度的西红花酸(1、0.1、0.01 μmol/L)预孵BEC细胞 12 h 后,用AGE(100 mg/L)刺激细胞 12 h, RT-PCR法测定RAGE mRNA的表达水平;ELISA法测定细胞间黏附分子-1(ICAM-1)的表达;试剂盒分别检测胞外超氧阴离子和硫代巴比妥酸反应产物(TBARS)浓度;同时,还用2,7-二氯荧光素(DCFH)测定了胞内H2O2的浓度,并用罗丹明123(Rh123)荧光法及MTT法分别检测细胞线粒体膜电位(MMP)水平和其琥珀酸脱氢酶(MSDH)的活性.结果:与AGE模型组相比,西红花酸能显著抑制RAGE mRNA的表达(P<0.05),降低胞外超氧阴离子和TBARS(P<0.01 或P<0.05)及胞内H2O2水平;结果还显示,西红花酸能提高细胞MMP水平和MSDH活性.对ICAM-1蛋白表达也有抑制作用,且呈时间和剂量依赖性.结论:西红花酸可能通过清除AGE与RAGE结合产生的活性氧(ROS)来抑制RAGE mRNA的高表达.提示西红花酸对糖尿病血管病变有潜在的治疗价值.
目的:研究西紅花痠對晚期糖基化終產物(AGE)誘導牛內皮細胞(BEC)中晚期糖基化終產物受體(RAGE)mRNA錶達的抑製作用,併探討其可能機製.方法:不同濃度的西紅花痠(1、0.1、0.01 μmol/L)預孵BEC細胞 12 h 後,用AGE(100 mg/L)刺激細胞 12 h, RT-PCR法測定RAGE mRNA的錶達水平;ELISA法測定細胞間黏附分子-1(ICAM-1)的錶達;試劑盒分彆檢測胞外超氧陰離子和硫代巴比妥痠反應產物(TBARS)濃度;同時,還用2,7-二氯熒光素(DCFH)測定瞭胞內H2O2的濃度,併用囉丹明123(Rh123)熒光法及MTT法分彆檢測細胞線粒體膜電位(MMP)水平和其琥珀痠脫氫酶(MSDH)的活性.結果:與AGE模型組相比,西紅花痠能顯著抑製RAGE mRNA的錶達(P<0.05),降低胞外超氧陰離子和TBARS(P<0.01 或P<0.05)及胞內H2O2水平;結果還顯示,西紅花痠能提高細胞MMP水平和MSDH活性.對ICAM-1蛋白錶達也有抑製作用,且呈時間和劑量依賴性.結論:西紅花痠可能通過清除AGE與RAGE結閤產生的活性氧(ROS)來抑製RAGE mRNA的高錶達.提示西紅花痠對糖尿病血管病變有潛在的治療價值.
목적:연구서홍화산대만기당기화종산물(AGE)유도우내피세포(BEC)중만기당기화종산물수체(RAGE)mRNA표체적억제작용,병탐토기가능궤제.방법:불동농도적서홍화산(1、0.1、0.01 μmol/L)예부BEC세포 12 h 후,용AGE(100 mg/L)자격세포 12 h, RT-PCR법측정RAGE mRNA적표체수평;ELISA법측정세포간점부분자-1(ICAM-1)적표체;시제합분별검측포외초양음리자화류대파비타산반응산물(TBARS)농도;동시,환용2,7-이록형광소(DCFH)측정료포내H2O2적농도,병용라단명123(Rh123)형광법급MTT법분별검측세포선립체막전위(MMP)수평화기호박산탈경매(MSDH)적활성.결과:여AGE모형조상비,서홍화산능현저억제RAGE mRNA적표체(P<0.05),강저포외초양음리자화TBARS(P<0.01 혹P<0.05)급포내H2O2수평;결과환현시,서홍화산능제고세포MMP수평화MSDH활성.대ICAM-1단백표체야유억제작용,차정시간화제량의뢰성.결론:서홍화산가능통과청제AGE여RAGE결합산생적활성양(ROS)래억제RAGE mRNA적고표체.제시서홍화산대당뇨병혈관병변유잠재적치료개치.
AIM:To investigate the effects of crocetin on receptor for advanced glycation end products(RAGE) mRNA expression in bovine endothelial cells(BEC) induced by advanced glycation end products(AGE) and the possible mechanisms involved. METHODS:The BEC were preincubated with crocetin(1,0.1 and 0.01 μmol/L) for 12 h,then exposed to AGE (100 mg/L). The RAGE mRNA expression was detected by RT-PCR analysis, the intercellular adhesion molecule-1(ICAM-1) was measured by ELISA. The extracellular superoxide ion and thiobarbituric acid reactive substances (TBARS) were assessed with superoxide ion kit and colorimetric assay, respectively. The intracellular H2O2 was also detected using the probe 2,7-dichlorofluorescein (DCFH). The mitochondrial membrane potential(MMP) and mitochodrial Succinate dehydrogenase(MSDH) were analyzed by the retention of rhodamine123 (Rh123) and MTT. RESULTS:Compared with AGE Group, crocetin was able to significantly reduce RAGE mRNA expression (P<0.05), decrease the content of super anion and TBARS in supernatant media (P<0.01 or P<0.05) and H2O2 in cells (P<0.05). Spontaneously, the MMP and the activity of MSDH were improved. Furthermore, the expression of ICAM-1 protein was suppressed by crocetin in a time-and dose-dependant manner. CONCLUSION:These results demonstrated that crocetin could inhibit RAGE over-expression in AGE-exposed BEC by suppressing the generation of reactive oxygen species active oxygen(ROS). This study suggested that crocetin might have beneficial effects in the treatment of diabetic vascular complications.
