中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2013年
3期
193-197
,共5页
钱中清%李新萍%王才智%何小燕%方芳
錢中清%李新萍%王纔智%何小燕%方芳
전중청%리신평%왕재지%하소연%방방
先兆子痫%脐静脉%内皮细胞%活性氧%钙%膜电位,线粒体
先兆子癇%臍靜脈%內皮細胞%活性氧%鈣%膜電位,線粒體
선조자간%제정맥%내피세포%활성양%개%막전위,선립체
Pre-eclampsia%Umbilical veins%Endothelial cells%Reactive oxygen species%Calcium%Membrane potential,mitochondrial
目的 探讨重度子痫前期患者血清对人脐静脉血管内皮细胞(HUVEC)造成的氧化性损伤.方法 (1)HUVEC进行如下分组处理:20％健康孕妇血清刺激(正常血清组)、20％重度子痫前期患者血清刺激(PE组)、20％重度子痫前期患者血清+3×103 U/ml过氧化氢酶共同刺激(PE+Cat组),并设置空白对照组,继续培养后,透射电镜观察细胞损伤情况,并用流式细胞仪检测凋亡相关蛋白APO2.7的表达情况.(2)激光共聚焦显微镜下,对HUVEC进行实时扫描的同时给予如下分组处理:正常血清组、PE组、PE+ Cat组、加入100 μmol/L H2O2(阳性对照组),分别检测胞内活性氧簇(ROS)、Ca2及线粒体膜电位水平.结果 (1)透射电镜下,PE组HUVEC的损伤明显,出现细胞膜脱落,并包裹细胞质形成自噬体,线粒体水肿,核固缩；但PE+ Cat组的HUVEC虽然线粒体有轻度水肿表现,但嵴清晰可见,细胞核形态基本正常,细胞损伤明显改善.PE组的HUVEC中APO2.7的表达率为(37.8±1.1)％,与空白对照组[(13.4±1.1)％]、正常血清组[(13.5±1.5)％]比较显著升高,差异均有统计学意义(P＜0.01)；而PE+ Cat组APO2.7表达率为(19.2±1.6)％,显著降低,与PE组比较,差异有统计学意义(P＜0.01).(2)激光共聚焦显微镜下PE组的HUVEC胞内ROS、Ca2荧光强度变化幅度分别为12.0±1.3、4.1±0.7,而正常血清组分别为1.1±0.4、0.6±0.4,PE+Cat组分别为1.5±0.5、0.9±0.5,PE组分别与正常血清组、PE+ Cat组比较,差异均有统计学意义(P＜0.01).PE组的HUVEC线粒体膜电位荧光强度在扫描120 s时有显著降低,而正常血清组、PE+Cat组变化不明显.结论 重度子痫前期患者血清通过促进ROS生成,增强Ca2超载水平和降低线粒体膜电位对血管内皮细胞造成氧化性损伤.
目的 探討重度子癇前期患者血清對人臍靜脈血管內皮細胞(HUVEC)造成的氧化性損傷.方法 (1)HUVEC進行如下分組處理:20％健康孕婦血清刺激(正常血清組)、20％重度子癇前期患者血清刺激(PE組)、20％重度子癇前期患者血清+3×103 U/ml過氧化氫酶共同刺激(PE+Cat組),併設置空白對照組,繼續培養後,透射電鏡觀察細胞損傷情況,併用流式細胞儀檢測凋亡相關蛋白APO2.7的錶達情況.(2)激光共聚焦顯微鏡下,對HUVEC進行實時掃描的同時給予如下分組處理:正常血清組、PE組、PE+ Cat組、加入100 μmol/L H2O2(暘性對照組),分彆檢測胞內活性氧簇(ROS)、Ca2及線粒體膜電位水平.結果 (1)透射電鏡下,PE組HUVEC的損傷明顯,齣現細胞膜脫落,併包裹細胞質形成自噬體,線粒體水腫,覈固縮；但PE+ Cat組的HUVEC雖然線粒體有輕度水腫錶現,但嵴清晰可見,細胞覈形態基本正常,細胞損傷明顯改善.PE組的HUVEC中APO2.7的錶達率為(37.8±1.1)％,與空白對照組[(13.4±1.1)％]、正常血清組[(13.5±1.5)％]比較顯著升高,差異均有統計學意義(P＜0.01)；而PE+ Cat組APO2.7錶達率為(19.2±1.6)％,顯著降低,與PE組比較,差異有統計學意義(P＜0.01).(2)激光共聚焦顯微鏡下PE組的HUVEC胞內ROS、Ca2熒光彊度變化幅度分彆為12.0±1.3、4.1±0.7,而正常血清組分彆為1.1±0.4、0.6±0.4,PE+Cat組分彆為1.5±0.5、0.9±0.5,PE組分彆與正常血清組、PE+ Cat組比較,差異均有統計學意義(P＜0.01).PE組的HUVEC線粒體膜電位熒光彊度在掃描120 s時有顯著降低,而正常血清組、PE+Cat組變化不明顯.結論 重度子癇前期患者血清通過促進ROS生成,增彊Ca2超載水平和降低線粒體膜電位對血管內皮細胞造成氧化性損傷.
목적 탐토중도자간전기환자혈청대인제정맥혈관내피세포(HUVEC)조성적양화성손상.방법 (1)HUVEC진행여하분조처리:20％건강잉부혈청자격(정상혈청조)、20％중도자간전기환자혈청자격(PE조)、20％중도자간전기환자혈청+3×103 U/ml과양화경매공동자격(PE+Cat조),병설치공백대조조,계속배양후,투사전경관찰세포손상정황,병용류식세포의검측조망상관단백APO2.7적표체정황.(2)격광공취초현미경하,대HUVEC진행실시소묘적동시급여여하분조처리:정상혈청조、PE조、PE+ Cat조、가입100 μmol/L H2O2(양성대조조),분별검측포내활성양족(ROS)、Ca2급선립체막전위수평.결과 (1)투사전경하,PE조HUVEC적손상명현,출현세포막탈락,병포과세포질형성자서체,선립체수종,핵고축；단PE+ Cat조적HUVEC수연선립체유경도수종표현,단척청석가견,세포핵형태기본정상,세포손상명현개선.PE조적HUVEC중APO2.7적표체솔위(37.8±1.1)％,여공백대조조[(13.4±1.1)％]、정상혈청조[(13.5±1.5)％]비교현저승고,차이균유통계학의의(P＜0.01)；이PE+ Cat조APO2.7표체솔위(19.2±1.6)％,현저강저,여PE조비교,차이유통계학의의(P＜0.01).(2)격광공취초현미경하PE조적HUVEC포내ROS、Ca2형광강도변화폭도분별위12.0±1.3、4.1±0.7,이정상혈청조분별위1.1±0.4、0.6±0.4,PE+Cat조분별위1.5±0.5、0.9±0.5,PE조분별여정상혈청조、PE+ Cat조비교,차이균유통계학의의(P＜0.01).PE조적HUVEC선립체막전위형광강도재소묘120 s시유현저강저,이정상혈청조、PE+Cat조변화불명현.결론 중도자간전기환자혈청통과촉진ROS생성,증강Ca2초재수평화강저선립체막전위대혈관내피세포조성양화성손상.
Objective To investigate oxidative damage effect of the serum of severe preeclamptic patients on human umbilical vein endothelial cells (HUVEC).Methods (1) HUVEC were randomly divided into 4 groups according to the following:blank group as control,normal group added 20％ normal sera of pregnant women,group PE added 20％ sera of severe preeclamptic patients,and group PE + Cat added 20％ sera of severe preeclamptic patients plus 3 x 103 U/ml catalase.After cultured for 24 hours,the injury morphology and APO2.7 expression of HUVEC were detected by transmission electron microscopy and flow cytometry respectively.(2) Under the real-time scanning by laser scanning confocal microscopy,HUVEC were randomly divided into 4 groups according to the following:control group added 100 μmol/L H2O2 as positive control,normal group,group PE,and group PE + Cat.HUVEC of each group was scanned for 120 seconds to determine levels of reactive oxidative species (ROS),calcium homeostasis,and mitochondria membrane potential.Results (1) Obvious injury morphology of HUVEC was observed in group PE,and it was obviously improved by catalase in group PE + Cat.Percentage of HUVEC expressed APO2.7 was (37.8 ± 1.1) ％ in group PE,which was significantly higher than (13.4 ± 1.1) ％ in blank group or (13.5 ± 1.5) ％ in normal group,but significantly lower than (19.2 ± 1.6) ％ in group PE + Cat (all P ＜ 0.01).(2) The fluorescence intensity curves of intracellular ROS and calcium showed slowly rising in group PE,but no obvious changes in normal group and PE + Cat.The values of ROS and calcium in group PE (12.0±1.3,4.1 ±0.7) were higher than those in normal group (1.1 ±0.4,0.6 ±0.4),but lower than those in group PE + Cat (1.5 ± 0.5,0.9 ± 0.5 ; all P ＜ 0.01).Conclusion The serum of severe preeclamptic patients caused oxidative damage on HUVEC by increasing intracellular ROS generation,calcium overload,and decreasing mitochondrial membrane potential.
