CAJ | 학술논문

目的：研究在持续性高浓度胰岛素与间断性高浓度胰岛素不同浓度胰岛素的条件培养液的作用下，体外培养的人脐静脉内皮细胞（HUECV304）分泌一氧化氮（NO）的相应变化，从而进一步探究间断性高浓度胰岛素血症导致动脉粥样硬化的原因及致病机制。方法本研究用体外培养的 HUECV304细胞作为试验对象，研究间断性高浓度胰岛素对HUECV304细胞NO分泌的影响。试验共分为4组，即含100nmol／L胰岛素条件培养液与1nmol／L胰岛素细胞培养液轮换培养的间断性高浓度胰岛素组，含1nmol／L胰岛素细胞培养液的正常浓度胰岛素组，含100nmol／L胰岛素细胞培养液的持续性高浓度胰岛素组、不含胰岛素细胞培养液的阴性对照组，各组每间隔8小时均更换新鲜条件培养液1次，总共作用72小时。内皮细胞培养液上清中NO的含量用硝酸还原酶法检测。结果含不同浓度的胰岛素条件培养液作用内皮细胞72小时后，正常浓度组、间断性高浓度胰岛素组与持续性胰岛素组细胞分泌NO均出现不同程度的增加，分别为对照组的2．21倍（P＜0．05），7．13倍（P＜0．001）和4．50倍（P＜0．001）；其中间断性高浓度胰岛素组内皮细胞分泌NO增加尤为明显，明显高于持续性高浓度胰岛素组内皮细胞分泌NO（P＜0．05）。结论间断性高浓度胰岛素相对于持续性高浓度胰岛素能增强人脐静脉内皮细胞NO的分泌。提示血管内皮细胞NO的过度合成可能是间断性高浓度胰岛素损伤血管内皮的功能、促进糖尿病的慢性血管并发症的产生及其发展的重要原因。
목적：연구재지속성고농도이도소여간단성고농도이도소불동농도이도소적조건배양액적작용하，체외배양적인제정맥내피세포（HUECV304）분비일양화담（NO）적상응변화，종이진일보탐구간단성고농도이도소혈증도치동맥죽양경화적원인급치병궤제。방법본연구용체외배양적 HUECV304세포작위시험대상，연구간단성고농도이도소대HUECV304세포NO분비적영향。시험공분위4조，즉함100nmol／L이도소조건배양액여1nmol／L이도소세포배양액륜환배양적간단성고농도이도소조，함1nmol／L이도소세포배양액적정상농도이도소조，함100nmol／L이도소세포배양액적지속성고농도이도소조、불함이도소세포배양액적음성대조조，각조매간격8소시균경환신선조건배양액1차，총공작용72소시。내피세포배양액상청중NO적함량용초산환원매법검측。결과함불동농도적이도소조건배양액작용내피세포72소시후，정상농도조、간단성고농도이도소조여지속성이도소조세포분비NO균출현불동정도적증가，분별위대조조적2．21배（P＜0．05），7．13배（P＜0．001）화4．50배（P＜0．001）；기중간단성고농도이도소조내피세포분비NO증가우위명현，명현고우지속성고농도이도소조내피세포분비NO（P＜0．05）。결론간단성고농도이도소상대우지속성고농도이도소능증강인제정맥내피세포NO적분비。제시혈관내피세포NO적과도합성가능시간단성고농도이도소손상혈관내피적공능、촉진당뇨병적만성혈관병발증적산생급기발전적중요원인。
Objectives To compare the effects of fluctuate and persistent high insulin media on the production of NO in cultured human umbilical endothelial cells line ( HUECV304 ) so as to explore the mechanism of intermittent hyperinsulinemia on diabetic vascular complication.Methods Human umbilical vein endothelial cells ( HUECV304) were cultivated for 72 hours to 0 nmol/L insulin(control group),1 nmol/L insulin (nomal insulin group), 100nmol/L alternating with 1nmol/L insulin per 8 hours (fluctu-ate high insulin group) and 100 nmol/L insulin ( persistent high insulin group) respectively.The product of NO was measured with nitrate reduction test.Results It increased the production of NO significantly,after HUECV304s were exposed to normal insulin, fluctuate high insulin and persistent high insulin for 72 hours.Compared with cells exposed to control group media.the production of NO were increased to 2.21-folds(P<0.05), 7.13-folds(P<0.001)and 4.50-folds(P<0.001)respectively.Furthermore, com-pared with it in persistent high insulin group there was a significant increase of NO production in fluctuate high insulin group ( P<0.05).Conclusions Compared with persistent high insulin fluctuate high insulin further enhanced the production of NO in HUECVs in vitro.Our data suggest that by affecting the production of excessive NO of HUECVs high insulin fluctuation may be in-volved in promoting the development of chronic vascular complication of diabetes.