CAJ | 학술논문

目的观察氧化苦参碱对肾综合征出血热病毒的体内与体外抑制作用.方法体外实验:以不对细胞产生毒性的氧化苦参碱剂量作为实验剂量,流行性出血热病毒76-118株作为病毒株,用3种方法处理Vero细胞:①药物处理48 h后,分别用10-1～10-6不同稀释度的出血热病毒攻击,24 h后换维持液；②分别用10-1～ 10-6不同稀释度的出血热病毒攻击24 h,再用药物处理48 h后换维持液；③将10-1～ 10-6不同稀释度的出血热病毒与药物同时处理细胞,48 h后换维持液.每一病毒稀释度处理4孔细胞,同时设4孔作为阳性对照,用间接免疫荧光实验判定药物对出血热病毒的抑制作用.体内实验:2周龄地鼠30只,体质量30 ～ 40g,按体质量分为实验组和对照组,每组15只,用100TCID50出血热病毒进行腹腔接种(0.1 ml/只)；第4天实验组每日腹腔注射1∶100稀释的氧化苦参碱0.1 ml/只,对照组每日注射等量生理盐水;10 d后取鼠肺制备切片,进行免疫荧光检查.结果体外实验证实,氧化苦参碱按1∶8稀释为安全剂量；病毒稀释度为10-4时,方法2、3分别与对照组比较,差异有统计学意义(z值均为-2.53,P均＜0.05),方法1与对照组比较,差异无统计学意义(z=5.36,P＞0.05)；病毒稀释度为10-1～10-3、10-5、10-6时,各种方法差异无统计学意义(z值分别为0.00、-0.32、-0.19、4.21、4.21,P均＞0.05).体内实验证实,实验组与对照组比较,差异有统计学意义(z=-3.85,P＜0.05).结论氧化苦参碱对肾综合征出血热病毒有明显的抑制作用.
목적 관찰양화고삼감대신종합정출혈열병독적체내여체외억제작용.방법 체외실험:이불대세포산생독성적양화고삼감제량작위실험제량,류행성출혈열병독76-118주작위병독주,용3충방법처리Vero세포:①약물처리48 h후,분별용10-1～10-6불동희석도적출혈열병독공격,24 h후환유지액；②분별용10-1～ 10-6불동희석도적출혈열병독공격24 h,재용약물처리48 h후환유지액；③장10-1～ 10-6불동희석도적출혈열병독여약물동시처리세포,48 h후환유지액.매일병독희석도처리4공세포,동시설4공작위양성대조,용간접면역형광실험판정약물대출혈열병독적억제작용.체내실험:2주령지서30지,체질량30 ～ 40g,안체질량분위실험조화대조조,매조15지,용100TCID50출혈열병독진행복강접충(0.1 ml/지)；제4천실험조매일복강주사1∶100희석적양화고삼감0.1 ml/지,대조조매일주사등량생리염수;10 d후취서폐제비절편,진행면역형광검사.결과 체외실험증실,양화고삼감안1∶8희석위안전제량；병독희석도위10-4시,방법2、3분별여대조조비교,차이유통계학의의(z치균위-2.53,P균＜0.05),방법1여대조조비교,차이무통계학의의(z=5.36,P＞0.05)；병독희석도위10-1～10-3、10-5、10-6시,각충방법차이무통계학의의(z치분별위0.00、-0.32、-0.19、4.21、4.21,P균＞0.05).체내실험증실,실험조여대조조비교,차이유통계학의의(z=-3.85,P＜0.05).결론 양화고삼감대신종합정출혈열병독유명현적억제작용.
Objective To study the effects of oxymatrine as inhibitor of hemorrhagic fever with renal syndrome virus (HFRSV) infection in vitro and in vivo.Methods In vitro studies,a dose of oxymatrine without cytotoxicity and 76-118 strain of HFRSV was taken to treat Vero cells in three ways:①After treated with oxymatrine for 48 h,Vero cells were attacked by HFRSV at dilution of 10-1 ～ 10-6,respectively for 24 h before changing to maintenance medium; ②Vero cells were first attacked by HFRSV of 10-1 ～ 10-6 dilution respectively,then oxymatrine was used for 48 h before changing to maintenance medium; ③Vero cells were attacked by HFRSV at dilution of 10-1 ～ 10-6 respectively,and meanwhile treated with oxymatrine for 48 h before changing to maintenance mcdium.Each dilution handled four porocytes,and four positive controls were set up at the same time.Indirect immunofluorescence assay (IFA) was performed to determine the inhibitory effect of oxymatrine in experimental group and positive control.In vivo studies,thirty 2-week-old hamsters,weighing about 30-40 g,were divided into experimental and control groups according to body weight,n =15.These aninals were inoculated intraperitoneally with HFRSV in 100TCID50(0.1 ml each); on days 4-13,0.1 ml of oxymatrine 1:100 were given to each hamster in experimental group daily by intraperitoneal injection,while the same amount of saline was given to the control ones.Lung tissue of hamsters was then dissected out to slice to be identified by immunofluorcscence stain.Results It was demonstrated that oxymatrine with the diluted fractions of 1:8 was safe in vitro.When the virus dilution of HFRSV was l0-4,compared with control groups,the differences were statistically significant in method 2 and 3 (z =-2.53,-2.53,all P ＜ 0.05),while no statistical significance in method 1 (z＝5.36,P＞ 0.05).When the virus dilution of HFRSV was 10-1 ～ 10-3,10-5,10-6,the differences were not statistically significant (z--0.00,-0.32,-0.19,4.21,4.21,all P ＞ 0.05).In vivo studies,compared with control group,the differences were statistically significant in experimental group (z =-3.85,P ＜ 0.05).Conclusion Oxymatrine significantly inhibites HFRSV.