【作者】 冯红丽；

【导师】 俞诗源；

【作者基本信息】 西北师范大学 ， 细胞生物学， 2009， 硕士

【摘要】 目的通过对受孕小鼠连续注射海洛因,探讨了海洛因对各发育期仔鼠及母鼠体重、行为、肺组织抗氧化防御系统中SOD活力、CAT活力和MDA含量、肺组织结构及KGF、c-Fos蛋白和Bax蛋白阳性表达的影响,旨在为海洛因对动物肺发育的影响研究提供基础资料。方法对48例受孕小鼠(第8d开始)实验组每天早晚分别注射0.2ml浓度为1.0g/L、1.5g/L和2.0g/L的海洛因溶液,对照组注射等量的生理盐水直到小鼠分娩,观察测量其行为和体重变化,采用比色法检测胚胎15 d、生后1 d、生后7 d、生后15 d的仔鼠和母鼠肺中SOD、CAT活力及MDA含量的变化,应用生物显微技术观察各发育期仔鼠和母鼠肺组织结构的变化,并用免疫组织化学染色和体视学半定量方法检测各发育期仔鼠和母鼠肺中KGF、c-Fos蛋白和Bax蛋白表达的变化。结果1.海洛因影响仔鼠和母鼠的行为活动和体重及仔鼠的胚胎发育,实验组中母鼠出现兴奋跑圈、蹿起、腹泻等症状,仔鼠出现了饮奶次数减少,尖叫和扭体次数增多等症状,同时各发育期仔鼠和母鼠的体重明显低于对照组,2.0g/L海洛因组则出现了死胎和畸形胎鼠。2.海洛因影响仔鼠和母鼠肺中抗氧化酶活性,海洛因对各发育期仔鼠和母鼠肺中SOD、CAT活力有显著的抑制作用(P <0.05),且抑制作用随海洛因浓度的增大而加大,但MDA含量则显著增加(P <0.05),海洛因浓度越大仔鼠和母鼠肺中MDA的含量就越高。3.海洛因影响仔鼠和母鼠肺组织结构,1.5g/L和2.0g/L海洛因组各发育期仔鼠和母鼠肺泡直径和肺泡隔厚度与对照组相比显著增加(P <0.05),1.0g/L海洛因组变化不明显(P >0.05)。4.海洛因影响仔鼠和母鼠肺中相关活性物质的表达,1.5g/L和2.0g/L海洛因组各发育期仔鼠和母鼠肺中KGF、c-Fos蛋白和Bax蛋白的阳性表达与对照组相比显著增强(P <0.01或P <0.05),1.0g/L海洛因组变化不明显(P >0.05),且随海洛因浓度的增强阳性表达增大。但随着发育进行,海洛因组仔鼠和母鼠肺中KGF、c-Fos蛋白和Bax蛋白的阳性表达减弱。结论1.海洛因对仔鼠和母鼠的行为、体重和胚胎发育的影响具有长时程效应。2.海洛因对各发育期仔鼠和母鼠肺组织中SOD活力、CAT活力有损伤作用,使发育中仔鼠及母鼠肺中脂质过氧化水平升高,导致了海洛因对肺组织的损伤。3.海洛因对各发育期仔鼠和母鼠的肺组织结构都有不同程度的损伤作用,且海洛因浓度越大对肺的损伤越显著。4.海洛因影响各发育期仔鼠和母鼠肺组织中KGF、c-Fos蛋白和Bax蛋白的阳性表达,引起肺组织细胞凋亡加剧,但随着发育持续这种影响减弱。5.海洛因对各发育期仔鼠和母鼠的肺都有不同程度的损伤作用,这可能与肺组织中SOD、CAT活力的下降和MDA含量的升高有关,同时也可能与KGF、c-Fos蛋白和Bax蛋白表达的增强有关。更多还原

【Abstract】 ObjectiveIn order to study the effects of heroin on weight,behaviour changes and enzymaticactivityofsuperoxidedismutase(SOD)andcatalase(CAT),andmaleicdialdehyde(MDA)content and histological structure and positive expression of KGF,c-Fos and Bax in thelungs of the developing filial mice and mother mice by separate injecting into pregnantmicewithdifferentconcentrationsofheroin,toprovidebasicdatafortheresearchonlungdevelopmentinanimals.Methods48 mother mice at the eighth day of pregnancy began to be separately injected withdifferent concentrations (1.0 g/L,1.5 g/L and 2.0 g/L) of heroin (0.2 ml) in theexperimental groups and the same amount of saline in the control group in the morningand afternoon until the birth of filial mice. Changes of weight and behaviour wereobserved and the enzymatic activity of SOD,CAT and MDA content were respectivelyanalyzed by colorimetry in the lungs of filial mice and mother mice at embryo 15 andpostnatal ages of 1 d, 7 d, 15 d,the lung structure were observed by microscope andexpression of KGF,c-Fos and Bax were measured by immunohistochemistry combinedwith stereological methods in the lungs of filial mice and mother mice at the differentdevelopmentalstages.Results1. Heroin influenced behaviour and weight of filial mice and mother mice andembryonic development of filial mice.Inthe experimental groups mother mice appearedsymptomssuchasexcitedrunning,jumping,diarrheaandfilialmiceappearedsymptomssuch as number decrease at the breast and screaming and writhing number increase,alsothe weight of filial mice and mother mice of the experimental groups was lower than thecontrol groups at the different developmental stages.2.0 g/Lheroin group appeared deadandabnormalfetalmice.2. Heroin affected theantioxidant enzymes activity in the lungs of of filial mice andmothermice,therewasasignificantinhibitionfortheSOD,CATactivity(P<0.05)withheroin,and the inhibitive effects were stronger with the concentration of heroin increasing,but the MDAcontent significantly increased (P < 0.05) and was higher withtheincreasedconcentrationofheroin.3. Heroin affected the lunghistological structure of filial mice and mother mice,thediameterofalveolousandthicknessofinteralveolarseptumoffilialmiceandmothermicesignificantlyincreased(P<0.05)withinjection1.5g/Land2.0g/Lherointhanthat ofthecontrol at the different developmental stages,while the diameter of alveolous andthickness of interalveolar septum didn’t significantly change with injection 1.0 g/Lheroin(P>0.05).4. Heroin affected the expression of inter related polypeptides in the lungs of the filialmiceandmothermice,thepositiveexpressionofKGF,c-FosandBaxinthelungsoffilialmiceandmothermicesignificantlyincreased(P<0.01orP<0.05)withinjection1.5g/Land2.0g/Lherointhanthatofthecontrolatthedifferentdevelopmentalstages,whilethepositive expression of KGF,c-Fos and Bax didn’t significantly change with injection 1.0g/Lheroin(P>0.05),andthepositive expressionwerestronger withthe concentrationofheroin increasing.However, during the following development, the positive expressionofKGF,c-FosandBaxdecreasedgraduallyinthelungsoffilialmiceandmothermiceoftheheroingroups.Conclusion1. The influence of heroin had long time-history effect for the behaviour and weightofdevelopingfilialmiceandmothermiceandembryonicdevelopment.2. The activities of SOD,CAT activity were injuryed in the lungs of filial mice andmother mice at the different developmental stages byheroin,and lipid peroxidation leveloflungswereincreasedduringdevelopmentofthefilialmiceandmothermice,thenlungcellsweredamagedbyheroin.3. The lung structure of filial mice and mother mice at the different developmentalstagesweredamagedindifferentdegreesbyheroin,andthedamageweresignificantwiththeconcentrationofheroinincreasing.4. Heroin affected positive expression of KGF,c-Fos and Bax in the lungs of thedevelopingfilialmiceandmothermiceatthedifferentdevelopmentalstages,whichledtothe lung apoptotic cell increase,but the influences were weakening during the followingdevelopmentofthefilialmice. 5. The lungs of filial mice and mother mice at the different developmental stageswere damaged in different degrees by heroin. This damage might be correlated with thelowactivitiesofSOD、CATandhighcontentofMDAandtheincreasesoftheexpressionintensityofKGF,c-FosandBaxinfilialmiceandmothermicelung.更多还原