中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
42期
3008-3011
,共4页
丁丽%张锦%张绍维%赵文洲
丁麗%張錦%張紹維%趙文洲
정려%장금%장소유%조문주
胰高血糖素样肽1%糖尿病,2型%人脐静脉内皮细胞%一氧化氮合酶
胰高血糖素樣肽1%糖尿病,2型%人臍靜脈內皮細胞%一氧化氮閤酶
이고혈당소양태1%당뇨병,2형%인제정맥내피세포%일양화담합매
Glucagon-like peptide 1%Diabetes mellitus,type 2%Human umbilical vein endothelial cells%Endothelial nitric oxide synthase
目的 观察胰高血糖素样肽1(GLP-1)(9-36)对人脐静脉内皮细胞(HUVEC)内皮型一氧化氮合酶(eNOS)的影响.方法 在正常葡萄糖(5.5 mmol/L)或高糖(16.8 mmol/L)条件下以5～5000 pmol/L GLP-1 (9-36)孵育HUVEC,硝酸还原酶法检测HUVEC的一氧化氮(NO)释放量、荧光法检测细胞内eNOS活性,Western印迹法检测HUVEC内eNOS 1177位丝氨酸磷酸化水平和总蛋白水平,实时定量反转录PCR检测eNOS mRNA水平.结果 正常葡萄糖条件下,50 ～5000 pmol/L GLP-1(9-36)组HUVEC释放NO均高于对照组[(22.2 ±2.6)μmol/L],其中5000 pmol/L GLP-1 (9-36)组为(41.6 ±8.1)μmol/L;细胞内eNOS活性均高于对照组(1.00 ±0.00),其中5000 pmol/L GLP-1(9-36)组为1.76 ±0.12.5000 pmol/L GLP-1 (9-36)组eNOS 1177位丝氨酸磷酸化水平、mRNA及总蛋白水平均显著高于对照组.高糖条件下,GLP-1(9-36)仍具有上述作用.结论 GLP-1(9-36)可上调HUVEC中eNOS的活性和表达,使NO释放增加,可能是其发挥心血管保护作用的机制之一.
目的 觀察胰高血糖素樣肽1(GLP-1)(9-36)對人臍靜脈內皮細胞(HUVEC)內皮型一氧化氮閤酶(eNOS)的影響.方法 在正常葡萄糖(5.5 mmol/L)或高糖(16.8 mmol/L)條件下以5～5000 pmol/L GLP-1 (9-36)孵育HUVEC,硝痠還原酶法檢測HUVEC的一氧化氮(NO)釋放量、熒光法檢測細胞內eNOS活性,Western印跡法檢測HUVEC內eNOS 1177位絲氨痠燐痠化水平和總蛋白水平,實時定量反轉錄PCR檢測eNOS mRNA水平.結果 正常葡萄糖條件下,50 ～5000 pmol/L GLP-1(9-36)組HUVEC釋放NO均高于對照組[(22.2 ±2.6)μmol/L],其中5000 pmol/L GLP-1 (9-36)組為(41.6 ±8.1)μmol/L;細胞內eNOS活性均高于對照組(1.00 ±0.00),其中5000 pmol/L GLP-1(9-36)組為1.76 ±0.12.5000 pmol/L GLP-1 (9-36)組eNOS 1177位絲氨痠燐痠化水平、mRNA及總蛋白水平均顯著高于對照組.高糖條件下,GLP-1(9-36)仍具有上述作用.結論 GLP-1(9-36)可上調HUVEC中eNOS的活性和錶達,使NO釋放增加,可能是其髮揮心血管保護作用的機製之一.
목적 관찰이고혈당소양태1(GLP-1)(9-36)대인제정맥내피세포(HUVEC)내피형일양화담합매(eNOS)적영향.방법 재정상포도당(5.5 mmol/L)혹고당(16.8 mmol/L)조건하이5～5000 pmol/L GLP-1 (9-36)부육HUVEC,초산환원매법검측HUVEC적일양화담(NO)석방량、형광법검측세포내eNOS활성,Western인적법검측HUVEC내eNOS 1177위사안산린산화수평화총단백수평,실시정량반전록PCR검측eNOS mRNA수평.결과 정상포도당조건하,50 ～5000 pmol/L GLP-1(9-36)조HUVEC석방NO균고우대조조[(22.2 ±2.6)μmol/L],기중5000 pmol/L GLP-1 (9-36)조위(41.6 ±8.1)μmol/L;세포내eNOS활성균고우대조조(1.00 ±0.00),기중5000 pmol/L GLP-1(9-36)조위1.76 ±0.12.5000 pmol/L GLP-1 (9-36)조eNOS 1177위사안산린산화수평、mRNA급총단백수평균현저고우대조조.고당조건하,GLP-1(9-36)잉구유상술작용.결론 GLP-1(9-36)가상조HUVEC중eNOS적활성화표체,사NO석방증가,가능시기발휘심혈관보호작용적궤제지일.
Objective To explore the effects of glucagon-like peptide 1 (GLP-1 (9-36)) on endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells (HUVEC).Methods HUVEC cultured under the conditions of normal glucose (5.5 mmol/L) or high glucose (16.8 mmol/L)were incubated with 5-5000 pmol/L GLP-1 (9-36).NO production was assayed by the nitrate reductase method.The eNOS activities were detected by NOS assay kit,p-eNOS (ser-1177) level and total eNOS protein level by Western blot and eNOS mRNA level by real-time reverse transcription-polymerase chain reaction (RT-PCR).Results Under normal glucose condition,NO productions from HUVEC of 50-5000 pmol/L GLP-1 (9-36) groups were significantly higher than the control ((41.6 ± 8.1) μmol/L vs (22.2± 2.6)μmol/L,P ＜0.05).So were eNOS activities in cells (1.76± 0.12 vs 1.00 ± 0.00,P ＜0.05).Compared with the control group,the levels of eNOS phosphorylation at ser-1177,mRNA and total protein were significantly elevated in the 5000 pmol/L GLP-1 (9-36) group.Under high glucose condition,GLP-1(9-36) retained all the above effects.Conclusion GLP-1 (9-36) can increase NO release,eNOS activity and expression in HUVEC.This may be one of the underlying mechanisms for the protective effects of GLP-1 (9-36) on cardiovascular system.
