CAJ | 학술논문

目的构建百草枯(paraquat,PQ)诱导的急性肺损伤细胞模型,探讨血管紧张素转化酶抑制剂卡托普利(captopril,CAP)对PQ中毒模型的保护作用.方法实验地点在武汉协和医院中心实验室,采用人脐静脉内皮细胞系(human umbilical vein endothelial cells,HUVECs),给予含不同浓度的PQ和CAP堵养基孵育HUVECs,制备实验模型,MTT法检测细胞活性,半数成活率判定模型成功与否.依据干预药物的不同确定实验分组:不予以药物干预为正常对照组(normal control group,NCG);给予含400μmol/LPQ的培养基制备PQ损伤组;在PQ损伤同时予以CAP(10μmol/L)干预的为CAP保护组;因CAP干预时间的不同,分CAPA,CAPB,CAPC.检测在PQ损伤24 h后检测培养幕上清内SOD、MDA;免疫组织化学染色法检测业细胞结构的变化;流式细胞仪检测细胞凋亡,数据采用(x±s)表示,应用SPSS 16.0软件行统计学分析.结果 MTT法榆测PQ作用浓度,时间和CAP各组数值,采用单因素方差分析,F值分别为56.734,172.025,P<0.01,成功构建了PQ模型及确定CAP干预浓度;与PQ组相比,CAP各组MDA含量降低(t分别为5.913,3.945,-3.426,P<0.01),而SOD含量升高(t分别为5.463,-2.292,-1.297,P<0.01);细胞色素C含量明显减少及CAP绀凋亡率降低.结论血管紧张素抑制剂CAP具有清除活性氧自由基作用,可拮抗PQ对HUVECs细胞的损伤作用,为PQ中毒的基础研究和临床治疗提供思考.
목적 구건백초고(paraquat,PQ)유도적급성폐손상세포모형,탐토혈관긴장소전화매억제제잡탁보리(captopril,CAP)대PQ중독모형적보호작용.방법실험지점재무한협화의원중심실험실,채용인제정맥내피세포계(human umbilical vein endothelial cells,HUVECs),급여함불동농도적PQ화CAP도양기부육HUVECs,제비실험모형,MTT법검측세포활성,반수성활솔판정모형성공여부.의거간예약물적불동학정실험분조:불여이약물간예위정상대조조(normal control group,NCG);급여함400μmol/LPQ적배양기제비PQ손상조;재PQ손상동시여이CAP(10μmol/L)간예적위CAP보호조;인CAP간예시간적불동,분CAPA,CAPB,CAPC.검측재PQ손상24 h후검측배양막상청내SOD、MDA;면역조직화학염색법검측업세포결구적변화;류식세포의검측세포조망,수거채용(x±s)표시,응용SPSS 16.0연건행통계학분석.결과 MTT법유측PQ작용농도,시간화CAP각조수치,채용단인소방차분석,F치분별위56.734,172.025,P<0.01,성공구건료PQ모형급학정CAP간예농도;여PQ조상비,CAP각조MDA함량강저(t분별위5.913,3.945,-3.426,P<0.01),이SOD함량승고(t분별위5.463,-2.292,-1.297,P<0.01);세포색소C함량명현감소급CAP감조망솔강저.결론 혈관긴장소억제제CAP구유청제활성양자유기작용,가길항PQ대HUVECs세포적손상작용,위PQ중독적기출연구화림상치료제공사고.
Objective To establish paraquat(PQ)induced acute lung injury models induced by paraquat (PQ), and to study the protective effects of angiotension converting enzyme inhibitors (ACEI) with eaptopril (CAP) on the PQ posioning. Method All experiments were made in the central laboratory of Union Hospital. Human umbilical vein endothelial cells (HUVECs) were incubated with different concentration of paraquat and captopril to establish experimental models. Half of cell survival rate detected with MTF assay to judge whether the model was successful or not. Three groups were divided according to the different drug. application: normal control group without any drug intervetion, PQ group exposed to paraqnat with the concentration of 400 μmol/L and CAP group additionally exposed to captopril with concentration of 10 μmoL/L, which was repartifioned to three groups (CAPA, CAPB and CAPC) according to the different intervention time. The supematant was collected to measure the concentration of malondialdehyde(MDA) and superoxide dismutase(SOD) after PQ injury for 24 hours, cy-tochrome C detected by immunocytochemistry and apoptosis detected by flow cytometry. Data were expressed as mean±standard error of the mean (x±s). Statistical 'analysis was carried out with the soft SPSS 16.0. Results MTr assay detected the concentration & time of PQ intervention and the data of CAP groups, using the single-re-season variance analysis, F value were 56.734,172.025, P < 0.01 respectively, thus to suecessfidly construct the PQ model and determine the concentration of CAP intervention groups. Relative to the PQ group, MDA as well as levels of cytochrome C of the CAP group were significantly decreased (t = 5.913,3.945,-3.426, P <0.01); while SOD were markedly increased (t = 5.463,-2.292,-1.297, P < 0.01). It also showed that captopril markedly decreased PQ induced the rate of HUVECs apoptosis, the percentagen of PQ group apoptosis was 46.1%, while CAP groups apoptnsis were 4.3 %, 9.2% and 17 % respectively. Conclusions Captopril has the ability to scavenge reactive oxygen species, and to protect against the paraquat induced lung toxicity on HUVECs, which pro-vide the infonnafion for basic research and clinical treatment of PQ posioning.