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ID:32353500
大小:11.27 MB
页数:65页
时间:2019-02-03
《epcs移植促进兔肝静脉型bcs肝静脉回流作用的实验研究》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、徐州医学院硕士学位论文第三部分EPCs移植促进兔肝静脉型BCS肝静脉回流作用的实验研究中文摘要目的探讨EPCs移植在兔肝静脉型BCS模型中改善肝静脉回流障碍的的作用机制。方法12只肝静脉型BCS模型兔(建模成功术后4周)随机分为EPCs移植组和对照组,每组各6只。EPCs移植组用5ml注射器抽取3ml浓度为(2~3)X106/mlEPCs的细胞悬液,超声引导下经门静脉途径缓慢输入。对照组经门静脉输入等体积的生理盐水。移植术后4周,所有兔均全麻下经CDUS检查肝静脉阻塞情况及肝脏回声变化,测量肝静脉、门静脉主干内径及平均血流速度、测算门静脉每分钟血流量;插管测定门静脉主干压
2、力;处死动物,采集靶肝静脉及其引流区内的肝脏组织标本,进行HE染色、CD34免疫组化染色,光镜下观察肝组织结构变化,将实验组各观察点结果与对照组相应结果进行比较。结果1.EPCs移植术后,所有兔均未出现腹腔内出血、死亡等情况。2.CDUS示:所有兔靶肝静脉内膜增厚,管腔均狭窄,EPCs移植组和对照组无统计学差异(尸>0.05)。EPCs移植组、对照组靶肝静脉引流区肝实质分别呈均匀稍高回声、不均匀稍高回声。移植术后4周,对照组门静脉平均流速、血流量分别为(10.88±1.23)cm/s、(131.45±14.88)ml/min,EPCs移植组门静脉平均流速、血流量分别为(1
3、2.75±1.08)cm/s、(158.13±17.66)ml/min(氏0.05);对照组门静脉压力为(13.50±1.05)cmH20,EPCs移植组为(12.17+0.75)cmH20(火0.05)。3.大体观察:对照组肝脏表面欠光滑,质地较硬,表现为暗红色;EPCs移植组肝脏表面光滑,质地较软,呈现粉红色。4.光镜下:对照组肝细胞肿胀,局灶性肝细胞萎缩、坏死,部分区域成纤维条索样改变,肝静脉壁不光整,内膜增厚;EPCs移植组肝细徐州医学院硕士学位论文胞肿胀程度较轻,门静脉分支血管增多,门脉血管沟通较明显,肝静脉壁光整;对照组肝静脉窦微血管密度计数(15.95±1.
4、43)/HP,EPCs移植组(23.94+2.00)/HP(P.5、eToinvestigatethemethodsofisolatingandculturingendothelialprogenitorcells(EPCs)fromrabbitbonemalTowandidentifythecellsinordertoreadyforthesubsequentexperiments.MethodsAdultandhealthyrabbitswereselected.Rabbitbonemarrowmononuclearcslls(MNCs)wereisolatedbyhistopaquedensity-gradientcentrifug6、ationandweresuspendedinendcIthelialbasalmediumsupplementedwithE国也.MVBulle妇陆tandthenthecellswereplatedonfibronectin-coatedculturedishes.Processofcellsgrowthwasobserved.EPCswereidentifiedbyDillabeledacety’latedlowdensityliprotein(DU—acLDL)andFITClabeledI难A—llectin(HTC-UEA-1).necellsshowedre7、dflurescencewereceilsphagocytizedacLDL,whilethosewithgreengluorescenewerecellsbindwith黝.j.ImmunoflurescencewereperformedtoanalyzetheexpressionofCD.34andCD.133withPE。CD34andFITC.CD133.Results1.Observationofeellmorphous:newisolatedMNCSwereroundandtransparent.Partsofth
5、eToinvestigatethemethodsofisolatingandculturingendothelialprogenitorcells(EPCs)fromrabbitbonemalTowandidentifythecellsinordertoreadyforthesubsequentexperiments.MethodsAdultandhealthyrabbitswereselected.Rabbitbonemarrowmononuclearcslls(MNCs)wereisolatedbyhistopaquedensity-gradientcentrifug
6、ationandweresuspendedinendcIthelialbasalmediumsupplementedwithE国也.MVBulle妇陆tandthenthecellswereplatedonfibronectin-coatedculturedishes.Processofcellsgrowthwasobserved.EPCswereidentifiedbyDillabeledacety’latedlowdensityliprotein(DU—acLDL)andFITClabeledI难A—llectin(HTC-UEA-1).necellsshowedre
7、dflurescencewereceilsphagocytizedacLDL,whilethosewithgreengluorescenewerecellsbindwith黝.j.ImmunoflurescencewereperformedtoanalyzetheexpressionofCD.34andCD.133withPE。CD34andFITC.CD133.Results1.Observationofeellmorphous:newisolatedMNCSwereroundandtransparent.Partsofth
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