中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2015年
7期
744-750
,共7页
罗丽%张丹%古妮娜%魏伏%陈晓迎%姜丽萍%蔡国强%刘景仑
囉麗%張丹%古妮娜%魏伏%陳曉迎%薑麗萍%蔡國彊%劉景崙
라려%장단%고니나%위복%진효영%강려평%채국강%류경륜
血管内皮细胞%EA.hy926细胞%钙%嗜铬粒蛋白A%Chromofungin%肿瘤坏死因子-α
血管內皮細胞%EA.hy926細胞%鈣%嗜鉻粒蛋白A%Chromofungin%腫瘤壞死因子-α
혈관내피세포%EA.hy926세포%개%기락립단백A%Chromofungin%종류배사인자-α
Vascular endothelial cells%EAhy926 cells%Calcium%Chromogranin A%Chromofungin%Tumor necrosis factor alpha
目的 探讨嗜铬粒蛋白A(chromogranin A,CGA)衍生多肽Chromofungin (CHR)对肿瘤坏死因子-α(TNF-α)诱导血管内皮细胞钙内流的影响.方法 以人脐静脉内皮细胞系EA.hy926细胞为研究对象,建立TNF-α刺激炎症模型,实验分为对照组、TNF-α组、10 nmol/LCHR+ TNF-α组、100 nmol/L CHR+TNF-α组和1 000 nmoL/L CHR+ TNF-α组.Transwell小室法检测EA.hy926细胞通透性,激光共聚焦法检测EA.hy926细胞内钙离子浓度([Ca2+]i)的变化.结果 与对照组比较,TNF-α组明显增加EA.hy926细胞通透性[(1.189±0.086)vs.(1.771±0.195),t=4.343,P=0.01];10 nmol/L、100 nmol/L和1 000 nmol/L CHR抑制TNF-α诱导的EA.hy926细胞通透性增加,其中10 nmol/L和100 nmol/L CHR显著降低TNF-α诱导的EA.hy926细胞高通透性[(1.771±0.195)vs.(1.315±0.134),t=3.403,P=0.042;(1.771 ±0.195)vs.(1.236±0.181),t=3.985,P=0.017],1 000 nmol/L CHR降低TNF-α诱导的EA.hy926细胞高通透性,差异无统计学意义[(1.771 ±0.195)vs.(1.411 ±0.255),t=2.679,P=0.126].10nmol/L、100 nmol/L和1 000 nmol/L CHR作用于EA.hy926细胞时细胞内Ca2+浓度无明显变化.TNF-α诱导EA.hy926细胞外Ca2+快速内流,各时间点细胞内Ca2+荧光强度F40、Ftl100与起始荧光值F130比较,差异具有统计学意义[荧光强度分别为:(41.497 ±3.788)vs.(56.193 ±3.082),F=196.129,P=0.000;(41.497±3.788)vs.(53.457±4.536),F=101.19,P=0.000],10nmol/L、100 nmol/L和1 000 nmol/L CHR能够抑制TNF-α引起的Ca2+快速内流.结论 CHR抑制TNF-α诱导的EA.hy926细胞通透性增加,这一作用可能是通过阻止TNF-α诱导的细胞外Ca2+内流来实现的.
目的 探討嗜鉻粒蛋白A(chromogranin A,CGA)衍生多肽Chromofungin (CHR)對腫瘤壞死因子-α(TNF-α)誘導血管內皮細胞鈣內流的影響.方法 以人臍靜脈內皮細胞繫EA.hy926細胞為研究對象,建立TNF-α刺激炎癥模型,實驗分為對照組、TNF-α組、10 nmol/LCHR+ TNF-α組、100 nmol/L CHR+TNF-α組和1 000 nmoL/L CHR+ TNF-α組.Transwell小室法檢測EA.hy926細胞通透性,激光共聚焦法檢測EA.hy926細胞內鈣離子濃度([Ca2+]i)的變化.結果 與對照組比較,TNF-α組明顯增加EA.hy926細胞通透性[(1.189±0.086)vs.(1.771±0.195),t=4.343,P=0.01];10 nmol/L、100 nmol/L和1 000 nmol/L CHR抑製TNF-α誘導的EA.hy926細胞通透性增加,其中10 nmol/L和100 nmol/L CHR顯著降低TNF-α誘導的EA.hy926細胞高通透性[(1.771±0.195)vs.(1.315±0.134),t=3.403,P=0.042;(1.771 ±0.195)vs.(1.236±0.181),t=3.985,P=0.017],1 000 nmol/L CHR降低TNF-α誘導的EA.hy926細胞高通透性,差異無統計學意義[(1.771 ±0.195)vs.(1.411 ±0.255),t=2.679,P=0.126].10nmol/L、100 nmol/L和1 000 nmol/L CHR作用于EA.hy926細胞時細胞內Ca2+濃度無明顯變化.TNF-α誘導EA.hy926細胞外Ca2+快速內流,各時間點細胞內Ca2+熒光彊度F40、Ftl100與起始熒光值F130比較,差異具有統計學意義[熒光彊度分彆為:(41.497 ±3.788)vs.(56.193 ±3.082),F=196.129,P=0.000;(41.497±3.788)vs.(53.457±4.536),F=101.19,P=0.000],10nmol/L、100 nmol/L和1 000 nmol/L CHR能夠抑製TNF-α引起的Ca2+快速內流.結論 CHR抑製TNF-α誘導的EA.hy926細胞通透性增加,這一作用可能是通過阻止TNF-α誘導的細胞外Ca2+內流來實現的.
목적 탐토기락립단백A(chromogranin A,CGA)연생다태Chromofungin (CHR)대종류배사인자-α(TNF-α)유도혈관내피세포개내류적영향.방법 이인제정맥내피세포계EA.hy926세포위연구대상,건립TNF-α자격염증모형,실험분위대조조、TNF-α조、10 nmol/LCHR+ TNF-α조、100 nmol/L CHR+TNF-α조화1 000 nmoL/L CHR+ TNF-α조.Transwell소실법검측EA.hy926세포통투성,격광공취초법검측EA.hy926세포내개리자농도([Ca2+]i)적변화.결과 여대조조비교,TNF-α조명현증가EA.hy926세포통투성[(1.189±0.086)vs.(1.771±0.195),t=4.343,P=0.01];10 nmol/L、100 nmol/L화1 000 nmol/L CHR억제TNF-α유도적EA.hy926세포통투성증가,기중10 nmol/L화100 nmol/L CHR현저강저TNF-α유도적EA.hy926세포고통투성[(1.771±0.195)vs.(1.315±0.134),t=3.403,P=0.042;(1.771 ±0.195)vs.(1.236±0.181),t=3.985,P=0.017],1 000 nmol/L CHR강저TNF-α유도적EA.hy926세포고통투성,차이무통계학의의[(1.771 ±0.195)vs.(1.411 ±0.255),t=2.679,P=0.126].10nmol/L、100 nmol/L화1 000 nmol/L CHR작용우EA.hy926세포시세포내Ca2+농도무명현변화.TNF-α유도EA.hy926세포외Ca2+쾌속내류,각시간점세포내Ca2+형광강도F40、Ftl100여기시형광치F130비교,차이구유통계학의의[형광강도분별위:(41.497 ±3.788)vs.(56.193 ±3.082),F=196.129,P=0.000;(41.497±3.788)vs.(53.457±4.536),F=101.19,P=0.000],10nmol/L、100 nmol/L화1 000 nmol/L CHR능구억제TNF-α인기적Ca2+쾌속내류.결론 CHR억제TNF-α유도적EA.hy926세포통투성증가,저일작용가능시통과조지TNF-α유도적세포외Ca2+내류래실현적.
Objective To investigate the influence of Chromogranin A (CGA) derived-peptide chromofungin (CHR) on calcium influx of vascular endothelial cells induced by tumor necrosis factor alpha (TNF-α).Methods Human umbilical vein endothelial cell line (EA.hy926) was employed as the research object.EA.hy926 cells inflammation model was built by stimulation of TNF-α.The experimental was composed of control group,TNF-α stimulation group,10 nmol/L CHR + TNF-α group,100 nmol/L CHR + TNF-α and 1 000 nmol/L CHR + TNF-α group.The monolayer permeability of EA.hy926 cells was measured by Transwell method.The change of the intracellular Ca2 + concentration ([Ca2 +];) in EA.hy926 cells was assessed by laser scanning confocal microscope.Results Compared with the control group,the permeability of EA.hy926 cells increased significantly in TNF-α group [(1.189 ±0.086) vs.(1.77 ±0.195),t =4.343,P =0.01].The concentrations of 10 nmol/L、100 nmol/L and 1 000 nmol/L CHR could inhibit the hyperpermeability induced by TNF-α in EA.hy926 cells;in that,10 nmol/L and 100 nmol/L CHR decreased significantly the hyperpermeability of EA.hy926 cells induced by TNF-α [(1.771 ±0.195) vs.(1.315±0.134),t=3.403,P=0.042;(1.771±0.195) vs.(1.236±0.181),t=3.985,P =0.017].And 1 000 nmol/L CHR decreased the hyperpermeability of EA.hy926 cells induced by TNF-α,but there was no statistically significant difference in comparison with 10 nmol/L and 100 nmol/ L [(1.771 ±0.195) vs.(1.411 ±0.255),t =2.679,P=0.126].The differem concentrations of 10 nmol/L,100 nmol/L and 1 000 nmol/L CHR couldn' t induce a significantly different intracellular Ca2+ concentrations in EA.hy926 cells.The exposure to 22 ng/mL TNF-αt could evoke a rapid Ca2+ entry in EA.hy926 cells,and the comparison of the Ca2 + fluorescence intensity at different intervals showed that F~0 and Ft100 were significantly different to initial fluorescence intensity Ft30 [(41.497 ±3.788) vs.(56.193 ±3.082),F=196.129,P=0.000;(41.497±3.788) vs.(53.457±4.536),F=101.19,P=0.000],and 10 nmol/L,100 nmol/L and 1 000 nmol/L CHR could inhibit TNF-α induced rapid Ca2+ influx.Conclusions CHR could inhibit the TNF-α induced hyperpermeability in EA.hy926 cells likely by blocking TNF-α evoking extracellular Ca2+ entry into cells.
