敌草快对大鼠胎仔动脉管的毒性作用及毒作用机理的研究

Research on The Toxic Effects of Diquat on Dutus Arteriosus of Rat Fetus and The Mechanism of Toxic Effects

本文主要研究除草剂敌草快对胎仔动脉管的毒性作用及毒性作用机理。方法:①以Wistar孕鼠作为研究对象,取妊娠19、20、21d的雌性Wistar大鼠各8只,不同妊娠期动物随机分为实验组和对照组,每组4只。实验组皮下注射7mg/kg敌草快,对照组皮下注射等量生理盐水。3h后处死动物,每只孕鼠取3只胎仔,观察动脉管收缩情况。结果,给予敌草快的胎仔动脉管直径明显小于对照组,由敌草快引起的胎仔动脉管收缩的临界期在胎龄19d和20d之间。本文还通过体外试验和体内试验的方法,研究敌草快对胎仔动脉管的毒性作用机理。②采用体外试验的方法,取绵羊精囊微粒体丙酮粉末0.5mg,1μmol/L牛血红素,50μmol/L(1-C14)花生烯酸,1mmol/L敌草快,使全容量为0.1ml,于24℃下,恒温1min后,加入乙醚:乙醇:1mol/L柠檬酸(30:40:1)混合液0.3ml,使之停止反应,再加0.5g无水Na2SO4,之后把上层50μl有机溶媒层标到二氧化硅凝胶F254玻璃板上,同时作为标品把PGB1也点到上述玻璃板上,通过乙醚:石油醚:乙酸(85:15:0.1)溶媒展开15cm。用紫外线灯标记PGB1的展开位置后,固定到X线胶片上进行48h自动放射照相,确定放射性生成物的展开位置,并抄写花生烯酸、PGG2和PGH2的展开部分,用液体同位素光谱仪测定各自的放射能。同时用0.01μmol/L～1mmol/L的消炎镇痛药(

The article mainly research the toxic effects of diquat on dutus arteriosus of rat fetus and the mechanism of toxic effects.Method:①Put gravid Wistar rat as research target .Eight gravid female Wistar rats on each day 19,20,21of gestation wererespectively divided randomly into two groups ,4 rats per group.In the experimental group, the rats were administered withdiquat by subcutaneous injection at the dose of 7.0 mg/kg. In the control group , the rats were administered with saline bysubcutaneous injection. The all animals were sacrificed 3h after injection .Three fetuseswere used for observing contraction ofductus arteriosus per gravid rat .Result: The internal diameter of ductus arteriosus of fetuses of rat dams treated with diquat wassignificantly decreased than that in the control group. The critical contraction time of ductus arteriosus of rat's futus induced bydiquat is between 19d and 20d. This article still research the mechanism of the Toxic effects of diquat on dutus arteriosus of ratfutus through in vitro and in vivo experiment .②By in vitro experiment, get 0.5mg powder from the microsome of sheepspermatophore,1μmol/L hemachrome of cattle ,50μmol/L(1-C14)arachidonic acid ,1mmol/L diquat. Ensure the volume is 0.1 mland hold it for 1min under 24℃.Then join 0.3ml mixture of B ether , ethyl alcohol and 1mol/L critic acid(30:40:1), stop thereaction and join 0.5g pyrotechnite . Then put 50μl organic solvent level on the silicagel F254 glass plate. At the mean time , tipthe PGB1 on the former glass then outspread 15cm in the mixture of B ether ,petroleum benzin, acetic acid(85:15:1). Afterpunctuating the position of PGB1, fix them on X ray pellicle for 48h and take auto radiation pictures. Confirm the outspreadingposition of radiation resultant and tanscribe the outspreading part of arachidonic acid , PGG2 and PGH2.Mensurate eachradioactivity with liquid spectro analysis of isotopes and do the same experiment with 0.01μmol/L～1mmol/L indomethacin,then compare them. Result: The creation of PGG2 and PGH2 has nothing to do with diquat . In other words, the creation of PGG2and PGH2 is almost equal to the control group.③by in vivo experiment , divided randomly 21d gravid Wistar rats into 2 groups,16 rats per group. The rats in the experiment group were administered with diquat by subcutaneous injection at a dose of 7.0 mg/kg .The rats in the control group were administered with saline by subcutaneous injection .Take 8 gravid rats on 3h after injection,dissect them afrer anaesthetizing by B ether and sample blood from abdomen arteriosus. Then get 4 fetuses per gravid rat andcut the arm pit, sample blood. The control group take the same way. Observe PGE2 content in plasm of the fetus. Result: Thein vivo experiment more prove the contraction of ductus arteriosus of rat's fetus induced by diquant isnot related to PGE2. Thatis to say, the contraction of ductus arteriosus for rat's fetus induced by diquat isn't accomplished by preventing the composionof PGE2. Conclusion: Diquat affects ductus arteriosus of rat fetus .And the contraction induced by diquat is not related to PGE2.That is to say, the mechanism of contraction induced by diquat is different from the mechanism of contraction induced byindomethacin.