安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
Acta Universitatis Medicinalis Anhui
2015年
10期
1373-1377
,共5页
IGF1%FoxO1%高糖%内皮细胞%细胞增殖%移行
IGF1%FoxO1%高糖%內皮細胞%細胞增殖%移行
IGF1%FoxO1%고당%내피세포%세포증식%이행
insulin-like growth factor 1%forkhead transcription factors%high glucose%endothelia cells%prolifera-tion%migration
目的 观察高糖下胰岛素样生长因子1 ( IGF1 )对高糖下人脐静脉内皮细胞( HUVEC)增殖和移行的影响,并探讨其可能影响机制. 方法 体外培养HUVEC,分为对照组、高糖组、高渗组、高糖 +IGF1 组和高糖 +IGF1 +AZD5363组,四甲基偶氮唑蓝( MTT)比色法检测内皮细胞增殖,Tran-swell小室观察细胞移行. 实时荧光定量PCR测定丝氨酸/苏氨酸蛋白激酶(Akt)及叉头转录因子1(FoxO1)的mRNA水平. 免疫组化法测定内皮细胞 Akt、p-Akt、FoxO1 和 p-FoxO1的蛋白表达,并行半定量分析. 结果 与对照组比较,高糖组细胞存活率、移行率明显降低,FoxO1 mRNA表达明显升高,FoxO1/p-FoxO1蛋白表达增加,Akt/p-Akt比值增加,差异均有统计学意义( P<0. 05 ). 与高糖组比较,高糖+IGF1组细胞存活率、移行率明显增加, FoxO1 mRNA的表达明显降低,FoxO1/p-FoxO1蛋白表达降低,Akt/p-Akt蛋白表达降低,差异均有统计学意义( P<0. 05 );高糖+IGF1 +AZD5363组细胞存活率未见明显差异,移行未增加, FoxO1 mRNA的表达明显降低( P<0. 05 ) , FoxO1/p-FoxO1 未见明显差异. 5组Akt mRNA的表达差异无统计学意义. 结论IGF1能改善高糖对内皮细胞增殖移行的抑制作用,其机制可能是IGF1激活Akt 进一步调节FoxO1的表达.
目的 觀察高糖下胰島素樣生長因子1 ( IGF1 )對高糖下人臍靜脈內皮細胞( HUVEC)增殖和移行的影響,併探討其可能影響機製. 方法 體外培養HUVEC,分為對照組、高糖組、高滲組、高糖 +IGF1 組和高糖 +IGF1 +AZD5363組,四甲基偶氮唑藍( MTT)比色法檢測內皮細胞增殖,Tran-swell小室觀察細胞移行. 實時熒光定量PCR測定絲氨痠/囌氨痠蛋白激酶(Akt)及扠頭轉錄因子1(FoxO1)的mRNA水平. 免疫組化法測定內皮細胞 Akt、p-Akt、FoxO1 和 p-FoxO1的蛋白錶達,併行半定量分析. 結果 與對照組比較,高糖組細胞存活率、移行率明顯降低,FoxO1 mRNA錶達明顯升高,FoxO1/p-FoxO1蛋白錶達增加,Akt/p-Akt比值增加,差異均有統計學意義( P<0. 05 ). 與高糖組比較,高糖+IGF1組細胞存活率、移行率明顯增加, FoxO1 mRNA的錶達明顯降低,FoxO1/p-FoxO1蛋白錶達降低,Akt/p-Akt蛋白錶達降低,差異均有統計學意義( P<0. 05 );高糖+IGF1 +AZD5363組細胞存活率未見明顯差異,移行未增加, FoxO1 mRNA的錶達明顯降低( P<0. 05 ) , FoxO1/p-FoxO1 未見明顯差異. 5組Akt mRNA的錶達差異無統計學意義. 結論IGF1能改善高糖對內皮細胞增殖移行的抑製作用,其機製可能是IGF1激活Akt 進一步調節FoxO1的錶達.
목적 관찰고당하이도소양생장인자1 ( IGF1 )대고당하인제정맥내피세포( HUVEC)증식화이행적영향,병탐토기가능영향궤제. 방법 체외배양HUVEC,분위대조조、고당조、고삼조、고당 +IGF1 조화고당 +IGF1 +AZD5363조,사갑기우담서람( MTT)비색법검측내피세포증식,Tran-swell소실관찰세포이행. 실시형광정량PCR측정사안산/소안산단백격매(Akt)급차두전록인자1(FoxO1)적mRNA수평. 면역조화법측정내피세포 Akt、p-Akt、FoxO1 화 p-FoxO1적단백표체,병행반정량분석. 결과 여대조조비교,고당조세포존활솔、이행솔명현강저,FoxO1 mRNA표체명현승고,FoxO1/p-FoxO1단백표체증가,Akt/p-Akt비치증가,차이균유통계학의의( P<0. 05 ). 여고당조비교,고당+IGF1조세포존활솔、이행솔명현증가, FoxO1 mRNA적표체명현강저,FoxO1/p-FoxO1단백표체강저,Akt/p-Akt단백표체강저,차이균유통계학의의( P<0. 05 );고당+IGF1 +AZD5363조세포존활솔미견명현차이,이행미증가, FoxO1 mRNA적표체명현강저( P<0. 05 ) , FoxO1/p-FoxO1 미견명현차이. 5조Akt mRNA적표체차이무통계학의의. 결론IGF1능개선고당대내피세포증식이행적억제작용,기궤제가능시IGF1격활Akt 진일보조절FoxO1적표체.
Objective To investigate the effects and the possible mechanism of insulin like growth factor-1 ( IGF1 ) on proliferation and migration of human umbilical vein endothelial cells ( HUVEC) in high glucose. Methods Cultured HUVEC were divided into 5 groups( control group, high glucose group, hypertonic group, high glucose+IGF 1 group, high glucose+IGF1+ AZD5363 group) . The proliferation was detected by methyl thiazolyl tetrazolium ( MTT) , the migration was detected by transwellchamber. The mRNA expressions of serine/threonine kinase( Akt) and FoxO1 were detected by real-time PCR. The protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were detec-ted by immunohistochemisty. Results Compared with the control group, the rate of cell survival in high glucose group decreased significantly(P<0. 05), the transitional was decreased significantly(P<0. 05), the mRNA ex-pression of FoxO1 was increased significantly( P<0. 05 ) , the protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were increased. Compared with high glucose group, the cell survival rate and migration rate in high glucose+IGF1 group were increased significantly, the mRNA expression of FoxO1 was decreased significantly(P<0. 05), the protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were decreased. Compared with high glucose group, the cell survival rate and migration rate in high glucose+IGF1+AZD5363 group had no significant difference, the mR-NA expression of FoxO1 was decreased significantly ( P<0. 05 ) , the protein expressions of FoxO1/p-FoxO1 had no significant difference. The mRNA expression of Akt in five groups had no significant difference. Conclusion IGF1 can improve the proliferation and migration of endothelial cell in high glucose. It shows that IGF1 acts through Akt to promote FoxO1 phosphorylation.
