Oxidative damage effect of the serum of severe preeclamptic patients on human umbilical vein endothelial cell / 中华妇产科杂志

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.

Detection of systematic oxidative stress in preeclampsia / 基础医学与临床

Objective To detect systematic oxidative stress in preeclampsia.Methods (1)Morphological features of placenta hypoxia were observed by histological method ; (2) Level of granulocyte intracellular reactive oxygen species was monitored by dyeing full blood with 2' ,7'-dichlorodihydrofluorescein diacetate (H2DCFDA) ; (3) Level of H_2O_2 in sera was detected by special kits.Results Compared to normal pregnancy,placentas from preeclampsia showed distinct features of hypoxic stress injury,such as more syncytial knots formation,fibrosis emerged,vein in-jury and loss its normal configuration; Fluorescence values of ROS probe in neutrophils from different women were 45.61±12.20(n =49),51.02 ± 13.60(n =56,P <0.01)and 85.10 ± 16.30(n =47,P <0.01); Concentra-tions of H_2O_2were (24.57±5.17)μmol/L(n =49),(26.61±3.25)μmol/L(n =56,P 0.01) and (39.84±9.67)μmol/L(n=47,P<0.01) respectively.Conclusion With the help of histological method,flow cytometry and special kits,systematic oxidative stress can be detected through checking placentic tissues,netrophils and sera of preeclampsia.

VEGF induces HUVECs to produce extracellular H_2O_2 and its proliferation role / 中国病理生理杂志

AIM: To study the effect of VEGF on extracellular H2O2 production in HUVECs and the role of H2O2 in the VEGF-induced proliferation. METHODS: HUVECs was stimulated with 500 ?g/L VEGF. Products of extracellular H2O2 was detected by H2DCFDA staining. MTT method was used to value the influences of 3?106 U/L catalase and 5-20 mmol/L H2O2 to VEGF function. RESULTS: After treatment for 15 min with VEGF, HUVECs appeared fluorescence, and continued to become stronger, peaked at 45 min then decreased. HUVECs, which was treated simultaneity with VEGF and 3?106 U/L catalase, only appeared very faint fluorescence. The proliferation of HUVECs by VEGF was restrained when treated with 3?106 U/L catalase. The extrinsic H2O2 at concentration of 5-10 mmol/L promoted the proliferation of HUVECs but inhibited the proliferation effect of VEGF on HUVECs (P

THE EFFECTS OF LASER IRRADIATION ON INTRACELLULAR ROS,CALCIUM CONCENTRATION AND CELL MEMBRANE INTEGRITY / 解剖学报

Objective To investigate the effects of laser irradiation on intracellular ROS(reactive oxidant species),intracellular calcium concentration(_i,and cell membrane integrity in the process of live cell imaging with confocal laser scanning microscopy. Methods The effects of a given laser irradiation on ROS,intracellular calcium concentration(_i and cell viability were revealed respectively by stained ECV-304 with H_2DCFDA,Fluo-4AM and calcein-AM/PI,and visualized and analyzed using ultra view LCI(live cell image)confocal microscopy. Results The irradiation of 488nm laser induced fluorescent intensity of DCF to increase abruptly and attain the climax in about 80 seconds,afterwards the fluorescent intensity fell and returned to the baseline.In the 70 minutes of the irradiation,the fluorescent intensity of intracellular Fluo-4 kept a slightly ascending tendency.The fluorescent intensity of calcein decreased 15minutes after the irradiation,and serval cells were PI positively stained.Conclusion 488nm laser irradiation induces intracellular reactive oxidant species(ROS) and calcium concentration to increase,but there is no significant influence on cell membrane integrity.