中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2012年
6期
365-369
,共5页
刘静%闫立萍%魏素虹%许衍甲
劉靜%閆立萍%魏素虹%許衍甲
류정%염립평%위소홍%허연갑
糖基化终末产物,高级%内吗啡肽%人脐静脉内皮细胞%一氧化氮%内皮索
糖基化終末產物,高級%內嗎啡肽%人臍靜脈內皮細胞%一氧化氮%內皮索
당기화종말산물,고급%내마배태%인제정맥내피세포%일양화담%내피색
目的 探讨内吗啡肽( EMs)对糖基化终末产物(AGEs)培养条件下人脐静脉内皮细胞(HUVEC)合成分泌一氧化氮(NO)、内皮型一氧化氮合酶(eNOS)、内皮素1(ET-1)的影响.方法 体外制备AGEs修饰的牛血清白蛋白(AGEs-BSA)；分别用AGEs-BSA、EMs(1×10-5、1×10-6、1×10-7、1×10-8mol/L EM1或EM2)±AGEs-BSA及EMs(1×10-5 mol/L EM1或EM2)±纳洛酮±AGEs-BSA共同干预HUVEC,同时以只加BSA的HUVEC作为阴性对照.噻唑蓝(MTT)法检测各组HUVEC存活率；放射免疫法检测各组NO、eNOS水平；酶联免疫吸附实验(ELISA)法测定各组ET-1表达量；实时荧光定量聚合酶链反应(RT-PCR)法检测各组eNOS、ET-1 mRNA的表达.采用单因素方差分析及t检验进行数据统计.结果 与AGEs-BSA组相比,1×10-5mol/L EM1干预组HUVEC存活率增加,NO分泌量减少[分别为0.89 ±0.05 vs 0.67±0.02和(14.8±1.4)vs(23.0±0.7) μmol/L,t值分别为9.86、13.18,均p＜0.05],ET-1产生及其mRNA表达降低[分别为(0.85±0.01)vs(0.99±0.01)μg/L和10.6±0.7 vs 25.6±1.6,t值分别为31.36、50.18,均P＜0.05],eNOS活性及其mRNA表达增加[分别为(2.53±0.20)vs(0.61 ±0.09) U/ml和0.35 ±0.00 vs 0.05±0.00,t值分别为21.50、16.80,均P＜0.05],EM1其他浓度组和EM2各浓度组上述各指标与AGEs-BSA组比较结果相似,差异亦均有统计学意义(t值为2.24～102.4,均P＜0.05)；纳洛酮组上述指标与AGEs-BSA组差异无统计学意义(t值为0.56～2.05,均P＞0.05),而与各浓度EM1或EM2干预组差异有统计学意义(t值为2.24 ～64.96,均P＜0.05).结论 EMs可提高AGEs-BSA条件下HUVEC的存活率,降低NO、ET-1浓度,提高eNOS浓度；纳洛酮可阻断EMs的上述作用,提示EMs是通过μ阿片受体发挥作用的.
目的 探討內嗎啡肽( EMs)對糖基化終末產物(AGEs)培養條件下人臍靜脈內皮細胞(HUVEC)閤成分泌一氧化氮(NO)、內皮型一氧化氮閤酶(eNOS)、內皮素1(ET-1)的影響.方法 體外製備AGEs脩飾的牛血清白蛋白(AGEs-BSA)；分彆用AGEs-BSA、EMs(1×10-5、1×10-6、1×10-7、1×10-8mol/L EM1或EM2)±AGEs-BSA及EMs(1×10-5 mol/L EM1或EM2)±納洛酮±AGEs-BSA共同榦預HUVEC,同時以隻加BSA的HUVEC作為陰性對照.噻唑藍(MTT)法檢測各組HUVEC存活率；放射免疫法檢測各組NO、eNOS水平；酶聯免疫吸附實驗(ELISA)法測定各組ET-1錶達量；實時熒光定量聚閤酶鏈反應(RT-PCR)法檢測各組eNOS、ET-1 mRNA的錶達.採用單因素方差分析及t檢驗進行數據統計.結果 與AGEs-BSA組相比,1×10-5mol/L EM1榦預組HUVEC存活率增加,NO分泌量減少[分彆為0.89 ±0.05 vs 0.67±0.02和(14.8±1.4)vs(23.0±0.7) μmol/L,t值分彆為9.86、13.18,均p＜0.05],ET-1產生及其mRNA錶達降低[分彆為(0.85±0.01)vs(0.99±0.01)μg/L和10.6±0.7 vs 25.6±1.6,t值分彆為31.36、50.18,均P＜0.05],eNOS活性及其mRNA錶達增加[分彆為(2.53±0.20)vs(0.61 ±0.09) U/ml和0.35 ±0.00 vs 0.05±0.00,t值分彆為21.50、16.80,均P＜0.05],EM1其他濃度組和EM2各濃度組上述各指標與AGEs-BSA組比較結果相似,差異亦均有統計學意義(t值為2.24～102.4,均P＜0.05)；納洛酮組上述指標與AGEs-BSA組差異無統計學意義(t值為0.56～2.05,均P＞0.05),而與各濃度EM1或EM2榦預組差異有統計學意義(t值為2.24 ～64.96,均P＜0.05).結論 EMs可提高AGEs-BSA條件下HUVEC的存活率,降低NO、ET-1濃度,提高eNOS濃度；納洛酮可阻斷EMs的上述作用,提示EMs是通過μ阿片受體髮揮作用的.
목적 탐토내마배태( EMs)대당기화종말산물(AGEs)배양조건하인제정맥내피세포(HUVEC)합성분비일양화담(NO)、내피형일양화담합매(eNOS)、내피소1(ET-1)적영향.방법 체외제비AGEs수식적우혈청백단백(AGEs-BSA)；분별용AGEs-BSA、EMs(1×10-5、1×10-6、1×10-7、1×10-8mol/L EM1혹EM2)±AGEs-BSA급EMs(1×10-5 mol/L EM1혹EM2)±납락동±AGEs-BSA공동간예HUVEC,동시이지가BSA적HUVEC작위음성대조.새서람(MTT)법검측각조HUVEC존활솔；방사면역법검측각조NO、eNOS수평；매련면역흡부실험(ELISA)법측정각조ET-1표체량；실시형광정량취합매련반응(RT-PCR)법검측각조eNOS、ET-1 mRNA적표체.채용단인소방차분석급t검험진행수거통계.결과 여AGEs-BSA조상비,1×10-5mol/L EM1간예조HUVEC존활솔증가,NO분비량감소[분별위0.89 ±0.05 vs 0.67±0.02화(14.8±1.4)vs(23.0±0.7) μmol/L,t치분별위9.86、13.18,균p＜0.05],ET-1산생급기mRNA표체강저[분별위(0.85±0.01)vs(0.99±0.01)μg/L화10.6±0.7 vs 25.6±1.6,t치분별위31.36、50.18,균P＜0.05],eNOS활성급기mRNA표체증가[분별위(2.53±0.20)vs(0.61 ±0.09) U/ml화0.35 ±0.00 vs 0.05±0.00,t치분별위21.50、16.80,균P＜0.05],EM1기타농도조화EM2각농도조상술각지표여AGEs-BSA조비교결과상사,차이역균유통계학의의(t치위2.24～102.4,균P＜0.05)；납락동조상술지표여AGEs-BSA조차이무통계학의의(t치위0.56～2.05,균P＞0.05),이여각농도EM1혹EM2간예조차이유통계학의의(t치위2.24 ～64.96,균P＜0.05).결론 EMs가제고AGEs-BSA조건하HUVEC적존활솔,강저NO、ET-1농도,제고eNOS농도；납락동가조단EMs적상술작용,제시EMs시통과μ아편수체발휘작용적.
Objective To discuss the effects of endomorphins（EMs)on synthesis and secretion of vasoactive substances in human umbilical vein endothelial cells（HUVECs）under advanced glycation end products（AGEs）injury conditions.Methods HUVECs were stimulated withbovine serum albumin （BSA）,AGEs-BSA,AGEs-BSA+EMs（1×10-5,1×10-7,and 1×10-8mol/L),AGEs-BSA+EMs（1×10-5mol/L)+naloxone,respectively.The HUVECs viability was measured by methiazolyte-trazolium(MTT)assay,contents of nitric oxide(NO) and endouthelial nitric oxide synthase (eNOS) were detected by coiorimetric analysis,level of endothelin-1(ET-1) was detected by enzyme linked immunoabsorbent assay (ELISA),expression of eNOS and ET-1 mRNA was measured by reverse transcription polymerase chain reaction(RT-PCR).Data were analyzed by one-way ANOVA and t test.Result compared with the cells incubated with AGEs-BSA,the cell viability was significantly enhanced in 1×10-5mol/L EM1 group(0.89±0.05 vs 0.67±0.02,t=9.86,P＜0.05),while secretion of NO,secretion and mNAN expression of ET-1 was significantly decreased ((14.8±1.4)μmol/L,(0.85±0.01)μg/L,10.6±0.7vs(23.0±0.7)μmol/L,(0.99±0.01)μg/L,25.6±1.6,respctively;t values were 13.18,31.36,and 50.18,all P＜0.05),mRNA exprssion and secretion of eNOS was significantly enhanced（0.35±0.00,（2.53±0.20)U/ml vs 0.05±0.00,(0.99±0.01)U/ml,respectively; t values were 16.80 and 21.50,both P＜0.05）.Similar results were found in other groups of EMs,and the differences were statically significant between the EMs and AGEs-BSA group（t=2.24 to 102.40,allP＜0.05).There were no significant differences in above-mentioned indices between the AGEs-BSA +EMs + naloxone and AGEs-BSA group(t=0.56 to 2.05,all P＞0.05),but signficant differences were found between AGEs-BSA +EMs+naloxone and EMs groups(t=2.24 to 64.96,all P＜0.05).Conciusions EMs has a certain protective effect on AGEs-BSA-induced injury in HUVEC,which can be blocked by naloxone.
