CAJ | 학술논문

目的证实马兜铃酸(从)能使血管内皮细胞(VEC)胞内钙离子([Ca2+]i)超载,致VEC损伤,而羟苯磺酸钙有拮抗作用.方法体外培养人脐静脉内皮细胞系(HUVEC),用马兜铃酸钠(AA-Na)和羟苯磺酸钙处理,设对照组、AA组、干预组三组.倒置显微镜及透射电镜观察细胞形态及超微结构改变,ELISA法测定细胞培养上清液中血栓调节蛋白(TM),FLuo-3钙离子探针检测HUVEC([Ca2+]i)浓度变化.结果与对照组相比,AA组TM值和平均([Ca2+]i)浓度明显升高(P<0.05).与从组相比,干预组羟苯磺酸钙浓度为25.50μM时,TM值和平均([Ca2+]i)浓度明显降低(P<0.05);与对照组相比,AA组细胞内质网池状扩张,线粒体嵴缺损.而干预组细胞内质网、线粒体形态均有改善.结论 AA-Na能使VEC钙超载,致内皮细胞破坏,TM增加,内质网和线粒体破坏;羟苯磺酸钙可以通过保护内质网、线粒体,减少VEC钙超载,保护VEC.
목적 증실마두령산(종)능사혈관내피세포(VEC)포내개리자([Ca2+]i)초재,치VEC손상,이간분광산개유길항작용.방법 체외배양인제정맥내피세포계(HUVEC),용마두령산납(AA-Na)화간분광산개처리,설대조조、AA조、간예조삼조.도치현미경급투사전경관찰세포형태급초미결구개변,ELISA법측정세포배양상청액중혈전조절단백(TM),FLuo-3개리자탐침검측HUVEC([Ca2+]i)농도변화.결과 여대조조상비,AA조TM치화평균([Ca2+]i)농도명현승고(P<0.05).여종조상비,간예조간분광산개농도위25.50μM시,TM치화평균([Ca2+]i)농도명현강저(P<0.05);여대조조상비,AA조세포내질망지상확장,선립체척결손.이간예조세포내질망、선립체형태균유개선.결론 AA-Na능사VEC개초재,치내피세포파배,TM증가,내질망화선립체파배;간분광산개가이통과보호내질망、선립체,감소VEC개초재,보호VEC.
Objective .To prove aristolochic (AA) caused vascular endothelial cells (VEC) injury via intracellular calcium overloa-ding and investigate the mechanism of calcium dobesilate antagonism. Methods Human umbilical vein endothelial cells (HUVEC) were cultured in vitro, and randomly divided into three groups: Control group, AA group, intervention group. Microscope and transmission elec-tron microscopy were used to observe changes of cell morphology and ultrastructure. ELISA method were applied to determine thrombomedu-lin (TM) in cell culture supernatant, fluorescent indicator FLuo-3/AM and intracellular calcium concentration ([Ca2+]. Results TM val-ue and average [Ca2+] i of AA group were significantly higher than that of control group (P < 0.05). Compared with the AA group, when the concentration of calcium dobesilate was 25 μM or 50 μM, TM value and average [Caz +] significantly decreased in intervention group (P < 0.05). Compared with control group, endoplasmic reticulum was pool expansion shaped, and mitochondrial cristae was absent in AA group cells. Endoplasmic reticulum and mitochondria patterns in the intervention group cells showed some improvement, compared with AA group. Conclusion AA induced VEC calcium overloading, 'I'M secretion and injury of endothelial ceils, endoplasmic reticulum and mito-chondria destruction. Dabesilate calcium could protect endoplasmic reticulum and mitochondria and reduce AA induced VEC calcium over-loading, and these could protect VEC.