两种异位心脏移植小鼠模型的建立和比较

目的比较腹部和颈部小鼠心脏移植模型的优缺点,探讨二次器官移植或双心脏移植研究的可行性和实用性动物模型。方法参照Ono法和Chen法并加以改进,建立了同系或者同种小鼠腹部和颈部异位心脏移植模型,同时对两种手术方式的成功率、手术时间、移植心脏存活时间和病理组织学进行了比较。结果腹部和颈部异位心脏移植手术成功率分别为86.7%和83.3%,两种手术方式在总手术时间、移植心脏存活时间和病理组织形态学检查上均无明显差异(P0.05)。结论在熟练掌握显微外科技术的基础上,两种异位心脏移植模型都能顺利建立,两者既可分开选用,又可结合运用,以适应不同实验需求。     

大鼠异位工作型心脏移植模型的建立

目的 建立大鼠腹腔异位工作型心脏移植动物模型.方法 Wistar大鼠30只,将供心主肺动脉与左房吻合,再将供心移植到受体腹部,将升主动脉与供心腹主动脉端侧吻合,使得左心房获得前负荷,成为工作型心脏.结果 成功建立大鼠腹腔异位工作型心脏移植模型,15例成功11例,手术成功率为73%.手术平均时间为(38.8±5.7)min.移植心脏获得长期存活,在28 d后通过超声观察可见供体心脏工作正常.结论 手术的关键是提高供心的摘取技术及受体的血管吻合技术,同时术后给与供心适当的按摩辅助也非常必要.此模型可以更好的用于移植免疫学的研究.     

小鼠腹部心脏移植模型的制作与改进

目的建立并改进小鼠腹部异位心脏移植模型,为器官移植研究提供技术支持。方法SPF级近交系小鼠112只进行心脏移植手术,在传统方法的基础上进行改进,观察改进后手术效果并分析其优势。结果手术成功率为89.28％。总手术时间（103.9±16.5）min,受体手术时间（73.0±7.9）min。改进的方法简化了操作步骤,降低了手术难度。结论改进的小鼠腹部异位心脏移植技术是一种简便、有效、成功率高的模型制作方法。     

大鼠腹腔异位心脏移植模型建立及改进

目的:建立一种快速有效的大鼠腹腔异位心脏移植模型。方法:采用SD大鼠作为受体,Wistar大鼠作为供体,行同种异位腹腔心脏移植,术后给以CsA5 mg/kg/d灌服,心脏移植手术方法采用改良的Ono术式,观察改良的腹腔异位心脏移植各步骤所需时间、术后成功率及主要并发症发生率。结果:共建立40只大鼠异位心脏移植模型,手术成功率92.5%。动脉吻合时间12.5±2.3min,静脉吻合时间12.3±1.5 min,供心缺血时间37±3.5 min,受体血管阻断时间34.2±2.6 min,总手术时间90.2±4.8 min,出现的主要并发症为出血和供心复跳失败(各占5%、2.5%)。结论:改进的大鼠腹部异位心脏移植技术是一种简便、快速、有效、成功率高的模型制作方法。     

大鼠腹腔内工作型心脏移植模型的改进

目的构建大鼠腹腔内工作型心脏移植模型,总结影响模型成功率的因素。方法 Brown Norway到Lewis大鼠心脏移植90例,其中预实验50例,正式实验40例。采用肺动脉和左房吻合、主动脉和受体腹主动脉吻合的方法建立工作型心脏移植,统计手术存活率和死亡原因,分析确保成功率的关键因素。结果术者通过练习,手术成功率稳定77.5%,供心冷缺血时间为(34±5)min,整个手术时间为(71±11)min。HE染色显示移植后发生了免疫反应,移植模型可靠。结论决定手术成功率的影响因素众多,其中主要因素有:合格的实验动物、供体心脏的保护、快速有效的血管缝合和术后动物护理。     

负载供者抗原的第三方树突状细胞延长大鼠移植心脏存活

目的观察负载供者抗原第三方未成熟DC对大鼠同种异体移植心脏存活期的影响。方法培养骨髓来源第三方大鼠未成熟DC,负载供者抗原,CTLA-4 Ig致耐受处理。建立大鼠心脏腹部移植模型,A组：心脏移植;B组：心脏移植,术前输注供者源未成熟DC;C组：心脏移植,术前输注第三方未成熟DC;D组：心脏移植,术前输注负载供者抗原致耐受第三方未成熟DC。分别行移植心病理、IFNγmRNA表达和移植心存活时间观察。结果D组的移植心病理改变轻,IFNγmRNA表达下调,移植心脏存活时间显著延长。结论负载供者抗原第三方致耐受未成熟DC能够诱导供者特异性免疫耐受,明显延长大鼠移植心脏存活时间。     

大鼠心脏移植急性排斥反应动物模型的建立

目的为探索器官移植后急性排斥反应的一种快速可靠的诊断方法。方法先暴露和游离受体的吻合段血管,再获取供心以缩短移植心脏冷缺血时间;保留供者的心脏及右上肺,将其胸主动脉与受者的腹主动脉间断端侧吻合;开放血流时,先开放远心端再间断开放近心端,同时按摩心脏至心脏搏动规律、有力,其色泽恢复红润,右上肺亦充盈良好。结果A组(n=20)成功16只,动脉吻合口出血2只、心脏复跳失败1只、吻合口狭窄1只;B组(n=20)成功17只,2只死于吻合口出血,1只死于吻合口狭窄。A组手术成功率为80%,B组为85%(P>0.05)。结论(1)先暴露并游离好受体血管吻合段,再获取供心,可以缩短移植心冷缺血时间。(2)边灌注边获取的方法利于快速而又完整的获取供心。(3)在切取和植入的过程中一定要注意低温保护。(4)利用腹主动脉段进行移植,使手术方便易行。左心做功的大鼠腹部心脏移植模型简单、安全、实用,因其左心参与循环而使血流动力学更接近生理状态,是进行心脏移植及血流动力学研究的较理想模型。     

大鼠局灶性脑缺血模型评价方法间关联性分析

目的分析评价线栓法制备大鼠大脑中动脉阻塞模型成功的三种方法之间的相关性,为评价该模型提供一种新的方法和标准。方法雄性SD大鼠共(30)只,随机分成假手术组(n=15)、模型组(n=15),参照Zea-Longa线栓法稍加改进制备局灶性脑缺血模型,于术前术后检测各组大鼠脑血流变化。以大鼠脑梗死面积率为金标准,分析该标准与脑血流变化之间的相关性。结果模型大鼠脑梗死面积率与脑血流变化量呈正相关,与行为学评分高低无关。结论大鼠脑血流量变化可作为评价局灶性脑缺血模型成功的标准。     

220次大鼠心脏移植的经验体会

作者采用Ono等创建的大鼠异位心脏移植模型,并作了部分改进,手术成功率达93.5％,手术时间和供心缺血时间缩短。本文着重介绍手术方法与经验体会。     

肝郁脾虚泄泻大鼠模型的建立及评价

目的采用多因素刺激方法建立吻合人类"肝郁脾虚泄泻"疾病特征的大鼠模型,并选取操作简单、结论实时、科学经济的评价指标。方法将幼年SD大鼠随机分为空白组和模型组,模型组采用母婴分离、束缚刺激、直肠醋酸刺激三种因素复合造模,选择不同时间测定大鼠体重变化、直肠敏感性、大鼠粪便分型积分及粪便含水量等指标,客观验证肝郁脾虚泄泻模型是否成功。结果模型大鼠出现食欲减退,饮水量增大,尿液减少的病理现象,部分大鼠出现活动增加、甚至躁狂。粪便积分在5~7分,粪便含水量远远高于正常组大鼠,而肠道敏感性显著增加。结论该模型较好的模拟了肝郁脾虚泄泻的临床表现,能满足该类疾病的相关研究。     

转化生长因子β1对心脏移植排斥反应中穿孔素和颗粒酶B表达的影响

目的探讨转化生长因子β1（TGF-β1）对大鼠心脏移植排斥反应中穿孔素和颗粒酶B表达的影响。方法采用大鼠颈部心脏移植模型,实验动物随机分为3组：同系移植组、异系移植组和TGF-β1组。于术后第5d取移植心脏,用逆转录聚合酶链式反应（RT-PCR）法观察穿孔素和颗粒酶B的表达情况。结果异系移植组穿孔素和颗粒酶B表达明显升高（与同系移植组相比,P〈0．01）。TGF-β1组穿孔素和颗粒酶B表达明显降低（与异系移植组相比,P〈0．01）。结论TGF-β1对心脏移植排斥反应的免疫抑制作用可能与其抑制穿孔素和颗粒酶B的表达有关。     

不同性别大鼠在2型糖尿病造模过程中的稳定性

目的研究不同性别大鼠在2型糖尿病造模过程中的成功率及模型的稳定性。方法高糖高脂饮食联合腹腔注射小剂量链脲佐菌素诱导建立雄、雌性大鼠2型糖尿病模型。成模后所有大鼠每周固定时间测血糖和体重。观察24周后,心脏穿刺取血,测定空腹血糖（FPG）、血清胰岛素（FINS）、HbA1c、甘油三脂（TG）、胆固醇（TC）、高密度脂蛋白-胆固醇（HDL-C）、低密度脂蛋白-胆固醇（LDL-C）。结果单纯高糖高脂饮食喂养,雄、雌性大鼠血糖与正常组无显著差异;STZ注射后,雄性大鼠血糖升高并逐渐平稳,而雌性需两次STZ注射,模型才比较稳定。实验结束时,雄、雌性糖尿病大鼠FPG、FINS、HOMA-IR以及TG、TC、LDL-C均显著升高,说明模型存在胰岛素抵抗和脂代谢紊乱。结论高糖高脂饲料加一次性小剂量链脲佐菌素腹腔注射,可成功建立雄性大鼠2型糖尿病模型,而同等剂量,雌性模型需两次STZ。雄、雌性糖尿病大鼠模型具有高血糖、脂质代谢紊乱和胰岛素抵抗特点。造模成功率及稳定性与性别有关,雄性大鼠较雌性大鼠成模率高,稳定性好,且耗时更短。     

Comparison of the tube leukocyte adherence inhibition test with the lymphocyte stimulation assay in a rat transplantation model.

The tube leukocyte adherence inhibition test (LAI) was analyzed in a heterotopic heart transplantation model in BN and Wag/Rij rats. Seven, fourteen, and twenty-two days after transplantation spleen cells of the recipient rat were tested for recognition of histocompatibility antigens in crude membrane extracts prepared from normal rat tissue. It was found that addition of fetal calf serum affected the cell adherence considerably and that there was a large difference in the percentage of adherent cells from different animals. In a comparative study the lymphocyte stimulation assay proved to be more sensitive and reproducible than the LAI in this rat system.     

MicroRNA and mRNA Signatures in Ischemia Reperfusion Injury in Heart Transplantation

Ischemia reperfusion (I/R) injury is an unavoidable event occurring during heart transplantation, leading to graft failures and lower long-term survival rate of the recipient. Several studies have demonstrated that microRNAs (miRNAs) are vital regulators of signalling pathways involved in I/R injury. The present study aims to quantify the altered expression levels of miRNA and mRNA upon I/R injury in a mouse heart transplantation model, and to investigate whether these miRNA can regulate genes involved in I/R injury. We performed heterotopic heart transplantation on mouse models to generate heart tissue samples with I/R and non-I/R (control). The expression levels of miRNAs as well as genes were measured in heart grafts by microarray and real time RT-PCR. miRNA alteration in cardiomyocytes exposed to hypoxia was also detected by qRT-PCR. We observed significant alterations in miRNA and gene expression profile after I/R injury. There were 39 miRNAs significantly downregulated and 20 upregulated up to 1.5 fold in heart grafts with I/R injury compared with the grafts without I/R. 48 genes were observed with 3 fold change and p<0.05 and 18 signalling pathways were enriched using Keggs pathway library. Additionally, hypoxia/reperfusion induced primary cardiomyocyte apoptosis and altered miRNA expression profiles. In conclusion, this is the first report on miRNA expression profile for heart transplantation associated with I/R injury. These findings provide us with an insight into the role of miRNA in I/R injury in heart transplantation.     

Vascularized bone allograft transplantation in a genetically defined rat model

A heterotopic subcutaneous model for experimental vascularized bone allograft transplantation has been presented. This model uses genetically defined rats and allows serial assessment of graft viability. The reliability of this model has been proven by successful isograft transplantation. This model was used to study the effect of matching at the major histocompatibility complex on vascularized bone allograft survival. Whereas grafts transplanted across a minor histocompatibility barrier survived until sacrifice, grafts transplanted across a major histocompatibility barrier were victims of an acute rejection process. This study, therefore, showed genetic disparity to be a critical determinant of vascularized bone allograft survival. It indicates that primary vascularized bone allografts are as susceptible to rejection as heart and kidney allografts. For these reasons, it can be anticipated that genetic matching will be important in clinical vascularized bone allograft transplantation. The model used in this study should be useful for obtaining further fundamental immunologic information concerning vascularized bone allograft transplantation.     

Evaluation of the hormonal function and histological features of heterotopic isogenic ovarian transplantation in rats

The purpose of the study was to determine the feasibility of preserving ovarian function after heterotopic transplantation by means of microvascular anastomosis of the transplanted vascular pedicles to a set of preselected vessels. Six groups of 10 Sprague-Dawley inbred rats were used in this study. Group I underwent bilateral ovariectomy operation and served as the ovariectomy control. Group II underwent bilateral ovariectomy followed by heterotopic isogenic ovarian implantation. Group III underwent bilateral ovariectomy and isogenic heterotopic ovarian transplantation by means of microvascular anastomosis. Group IV served as the laparotomy sham-operated control. Group V served as the ovarian donor for group II. Group VI served as the donor of the ovarian-kidney vascular pedicle complex for group III. Postoperative ovarian estradiol levels were measured, and histological characteristics were elucidated in groups I, II, III, and IV. The results demonstrated that the estradiol level of the transplantation group was comparable to that of the sham operation group and was significantly higher than that of the implantation group. Histologically normal ovarian architecture was observed in the sham group (IV) and also in the transplantation group (III). Altered architecture was observed in the implantation group (II). These findings indicate that extraabdominal heterotopic ovarian transplantation with microvascular anastomosis led to normal ovarian hormonal function and was effective in preserving oocyte production capacity.