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JNK/c-Jun信号通路通过调控细胞骨架重构参与大鼠垂体柄损毁后精氨酸加压素神经元修复

发布时间:2024-05-21 01:19
  目的探究大鼠垂体柄损毁后精氨酸加压素(AVP)神经元通过双皮质素(DCX)促进神经元骨架修复的机制。方法取32只SD大鼠,随机分为实验组(损毁后1 d组,n=4;损毁进神经元骨架恢后3 d组,n=7;损毁后7 d组,n=7;损毁后14 d组,n=7)和对照组(假手术组,n=7),经颅顶损毁垂体柄,记录尿量、饮水量和尿比重,通过免疫荧光染色比较各组视上核(SON)AVP神经元中Ⅲ型β微管蛋白(Tuj1)、DCX和caspase3表达情况,并通过蛋白质印迹法探讨其调控机制。结果垂体柄损毁后,饮水量、尿量和尿比重出现了典型的三相性尿崩症(损毁后1~2 d为第一相,术后3~5 d为第二相,6 d以后为第三相)。免疫荧光的结果提示了AVP神经元损伤后修复现象,主要表现为DCX表达在损毁后不同程度的升高。AVP神经元中DCX阳性率分别为:假手术组,(37.93±1.83)%;损毁后1 d,(44.67±1.79)%;损毁后3 d,(48.09±2.96)%;损毁后7 d,(71.34±2.18)%;损毁后14 d,(61.08±2.15)%,(P<0.01)。同时,神经元骨架蛋白Tuj1也呈...

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图1垂体柄损毁后三相性尿崩症1201008060402001234567891011121314Urinevolume(mL/d)

图1垂体柄损毁后三相性尿崩症1201008060402001234567891011121314Urinevolume(mL/d)

低比重尿(1.006±0.001)和多饮(149±49mL/24h)。第3天时进入少尿期,表现少尿(20±12mL/24h),尿比重(1.020±0.007)升高,少饮(62±34mL/24h)。接近7d时,大鼠再次进入尿崩状态,表现为多尿(47±39mL/24h),尿比重(1.....


图2术后正中隆起和垂体柄处AVP神经纤维的矢状切片免疫组化提示轴突再生Fig.2ImmunohistochemistryofAVP-positivefibersinthesagittalsectionthroughtheratmedianeminenceandpituitarystalkafterPELshowingevidenceoftheregenerationofaxons.A:EarlystageafterPEL;B:Thelongterm28dstageafterPEL.Scalebars:500μm.3V,Thirdventricle;APit,anteriorlobeofpituitary;InfS,infundibularstem;IPit,intermediatelobeofpituitary;ME,medianeminence;PPit,

图2术后正中隆起和垂体柄处AVP神经纤维的矢状切片免疫组化提示轴突再生Fig.2ImmunohistochemistryofAVP-positivefibersinthesagittalsectionthroughtheratmedianeminenceandpituitarystalkafterPELshowingevidenceoftheregenerationofaxons.A:EarlystageafterPEL;B:Thelongterm28dstageafterPEL.Scalebars:500μm.3V,Thirdventricle;APit,anteriorlobeofpituitary;InfS,infundibularstem;IPit,intermediatelobeofpituitary;ME,medianeminence;PPit,

图2术后正中隆起和垂体柄处AVP神经纤维的矢状切片免疫组化提示轴突再生Fig.2ImmunohistochemistryofAVP-positivefibersinthesagittalsectionthroughtheratmedianeminenceandpituitarys....


图3垂体柄损毁远期出现神经元骨架修复现象Fig.3CytoskeletonrepairafterPEL.A:ImmunofluorescentstainingforTuj1(red)andAVP(green)ineachgroupinthesupraopticnucleus(SON)(n=4);B:QuantitativeanalysisofTuj1expressioninAVPneurons.Scalebars:100μmforlowmagnificationimage,15μmforhighmagnificationimage.*P<0.05,**P<0.01vsshamgroup.Postoperativedays

图3垂体柄损毁远期出现神经元骨架修复现象Fig.3CytoskeletonrepairafterPEL.A:ImmunofluorescentstainingforTuj1(red)andAVP(green)ineachgroupinthesupraopticnucleus(SON)(n=4);B:QuantitativeanalysisofTuj1expressioninAVPneurons.Scalebars:100μmforlowmagnificationimage,15μmforhighmagnificationimage.*P<0.05,**P<0.01vsshamgroup.Postoperativedays

图2术后正中隆起和垂体柄处AVP神经纤维的矢状切片免疫组化提示轴突再生Fig.2ImmunohistochemistryofAVP-positivefibersinthesagittalsectionthroughtheratmedianeminenceandpituitarys....


图4垂体柄损毁后DCX阳性率逐渐升高Fig.4ExpressionofDCXinAVPneurongraduallyincreasedafterPEL.A:ImmunofluorescentstainingforDCX(red)andAVP(green)inthesupraopticnucleus(SON)ineachgroup(n=4);B:QuantitativeanalysisofDCXexpressioninAVPneurons.Scalebars:100μmforlowmagnificationimage,15μmforhighmagnificationimage.*P<0.05,**P<0.01vsshamgroup.Postoperatived

图4垂体柄损毁后DCX阳性率逐渐升高Fig.4ExpressionofDCXinAVPneurongraduallyincreasedafterPEL.A:ImmunofluorescentstainingforDCX(red)andAVP(green)inthesupraopticnucleus(SON)ineachgroup(n=4);B:QuantitativeanalysisofDCXexpressioninAVPneurons.Scalebars:100μmforlowmagnificationimage,15μmforhighmagnificationimage.*P<0.05,**P<0.01vsshamgroup.Postoperatived

榻峁?允荆珼CX在正常的AVP神经元中也存在表达,而目前大多数观点认为DCX大多在未成熟的神经元中表达[14-15]。在关于DCX超微结构的报道中[8],DCX与神经元驱动蛋白3关系密切,在神经元囊泡运输中具有重要作用,AVP神经元中丰富的细胞骨架具有强大的运输能力,DCX在正常....



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