中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2014年
2期
142-145
,共4页
刘多谋%黄鹤光%周武汉%陈志耀%陆逢春
劉多謀%黃鶴光%週武漢%陳誌耀%陸逢春
류다모%황학광%주무한%진지요%륙봉춘
毛细血渗漏综合征%血管内皮屏障%白细胞介素-1β%水通道蛋白-1%细胞凋亡
毛細血滲漏綜閤徵%血管內皮屏障%白細胞介素-1β%水通道蛋白-1%細胞凋亡
모세혈삼루종합정%혈관내피병장%백세포개소-1β%수통도단백-1%세포조망
Capillary leak syndrome%Vascular endothelial barrier%Interleukin-1β%Aquaporin-1%Apoptosis
目的 观察白细胞介素-1β(IL-1β)作用下血管内皮细胞水通道蛋白-1(AQP-1)表达、凋亡和超微结构的改变,探讨IL-1β在毛细血管渗漏综合征中的作用.方法 体外培养脐静脉内皮细胞,随机分为3组:时间组:以含20 μg/L IL-1β的培养液分别培养3 h(T1)、8 h(T2)、12 h(T3)和24 h(T4)；浓度组:分别以含0.2μg/L(C1)、2μg/L(C2)和20μg/L(C3) IL-1β的培养液培养24h；对照组,以不含IL-1β的培养液培养24 h.实时荧光定量PCR和蛋白印迹检测AQP-1 mRNA和蛋白表达改变,流式细胞仪检测细胞凋亡情况,电镜观察细胞超微结构的改变.结果 与对照组相比,时间组T1 ～T4和浓度组C1～C3的AQP-1mRNA和蛋白表达均下降(P＜0.05).IL-1β刺激后,细胞凋亡率增高,电镜下可见线粒体肿胀、空泡变性、细胞核溶解及细胞坏死.而且,随着刺激时间的延长或浓度的增高,这些改变更加明显.结论 IL-1β可导致AQP-1mRNA和蛋白表达降低,细胞凋亡率增高,破坏细胞的超微结构.这是血管内皮屏障损伤的重要原因,可能与毛细血管渗漏综合征的发生密切相关.
目的 觀察白細胞介素-1β(IL-1β)作用下血管內皮細胞水通道蛋白-1(AQP-1)錶達、凋亡和超微結構的改變,探討IL-1β在毛細血管滲漏綜閤徵中的作用.方法 體外培養臍靜脈內皮細胞,隨機分為3組:時間組:以含20 μg/L IL-1β的培養液分彆培養3 h(T1)、8 h(T2)、12 h(T3)和24 h(T4)；濃度組:分彆以含0.2μg/L(C1)、2μg/L(C2)和20μg/L(C3) IL-1β的培養液培養24h；對照組,以不含IL-1β的培養液培養24 h.實時熒光定量PCR和蛋白印跡檢測AQP-1 mRNA和蛋白錶達改變,流式細胞儀檢測細胞凋亡情況,電鏡觀察細胞超微結構的改變.結果 與對照組相比,時間組T1 ～T4和濃度組C1～C3的AQP-1mRNA和蛋白錶達均下降(P＜0.05).IL-1β刺激後,細胞凋亡率增高,電鏡下可見線粒體腫脹、空泡變性、細胞覈溶解及細胞壞死.而且,隨著刺激時間的延長或濃度的增高,這些改變更加明顯.結論 IL-1β可導緻AQP-1mRNA和蛋白錶達降低,細胞凋亡率增高,破壞細胞的超微結構.這是血管內皮屏障損傷的重要原因,可能與毛細血管滲漏綜閤徵的髮生密切相關.
목적 관찰백세포개소-1β(IL-1β)작용하혈관내피세포수통도단백-1(AQP-1)표체、조망화초미결구적개변,탐토IL-1β재모세혈관삼루종합정중적작용.방법 체외배양제정맥내피세포,수궤분위3조:시간조:이함20 μg/L IL-1β적배양액분별배양3 h(T1)、8 h(T2)、12 h(T3)화24 h(T4)；농도조:분별이함0.2μg/L(C1)、2μg/L(C2)화20μg/L(C3) IL-1β적배양액배양24h；대조조,이불함IL-1β적배양액배양24 h.실시형광정량PCR화단백인적검측AQP-1 mRNA화단백표체개변,류식세포의검측세포조망정황,전경관찰세포초미결구적개변.결과 여대조조상비,시간조T1 ～T4화농도조C1～C3적AQP-1mRNA화단백표체균하강(P＜0.05).IL-1β자격후,세포조망솔증고,전경하가견선립체종창、공포변성、세포핵용해급세포배사.이차,수착자격시간적연장혹농도적증고,저사개변경가명현.결론 IL-1β가도치AQP-1mRNA화단백표체강저,세포조망솔증고,파배세포적초미결구.저시혈관내피병장손상적중요원인,가능여모세혈관삼루종합정적발생밀절상관.
Objective To explore the role of IL-1β in capillary leak syndrome by observing the alterations of AQP-1 expression,apoptosis,and ultrastructural of vascular endothelial cells under the action of IL-1β.Methods Umbilical vein endothelial cells (UVEC) in vitro were randomly allocated into 3 groups:time,concentration,and control.In the time group,UVECs were treated with culture medium containing 20 μg/L IL-1β for3 h(T1),8 h(T2),12 h(T3) and 24 h(T4).In the concentration group,UVECs were treated with culture medium containing 0.2 μg/L(C1),2 μg/L(C2) and 20 μg/L(C3) IL-1β for 24 h.In the control group,UVECs were treated with culture medium without IL-1β for 24 h.The changes of AQP-1 mRNA and protein expression were detected by real-time PCR and Western blot.Apoptosis was detected by flow cytometry,and cell ultrastructural changes were observed by electron microscopy.Results AQP-1 mRNA and protein expression of T1-T4 in the time group and C1-C3 in the concentration group were lower than those of the control group (P ＜ 0.05).The apoptotic rate was increased,and mitochondrial swelling,vacuolar degeneration,karyolysis and necrosis were observed under electron microscopy.These were more pronounced with time or concentration increases.Conclusions IL-1β can cause a decrease of AQP-1 mRNA and protein expression,increase in apoptotic rate and increase in damage to the cells'ultrastructure.This is an important reason for damage to the vascular endothelial barrier and may be associated with capillary leak syndrome.
