CN110338149A - 一种兔vx2肝癌alpps手术模型的建立方法 - Google Patents
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Abstract
本发明涉及医学领域,具体为一种兔VX2肝癌ALPPS手术模型的建立方法。其步骤如下:(1)采用瘤组织块接种法制作VX2兔肝癌模型。(2)经CT扫描观察,筛选符合尺寸要求的模型,实验组荷瘤兔行ALPPS一期手术。本发明是首次在兔VX2肝癌上建立ALPPS手术模型。兔VX2肿瘤是肝癌成像研究较为理想的实验模型,与人类肝细胞癌的血供特点和影像学表现非常相似,对临床具有指导作用。而在临床工作中上,ALPPS手术适用于中晚期肝癌患者。因此本发明构建的兔VX2肝癌ALPPS手术模型可以对肿瘤及残余正常肝脏进行整体性的研究。
Description
技术领域
本发明涉及医学领域,具体为一种兔VX2肝癌ALPPS手术模型的建立方法。
背景技术
在现有技术中,肝癌是中国发病率和死亡率最高的癌症之一,手术切除是目前肝癌最主要的治疗方式。然而中晚期肝癌患者,常因术后残余肝脏的体积(future liverremnant,FLR)不够代偿,术后肝脏功能衰竭风险较大,而丧失了手术切除治疗的可能。2012年,德国的Schnitzbauer首次报道了联合离断和门静脉结扎的二步肝切除术(AssociatingLiver Partition And Portal Vein Ligation For Staged Hepatectomy,ALPPS)。该术式既可以把肿瘤入肝血流全部阻断,又可以把两个健侧肝脏和带瘤肝脏之间可能存在的侧支循环全部阻断,从而在短时间内有效地诱导刺激残余肝脏体积的代偿性增大,为第二次手术创造更好的条件,进一步提高肝癌根治性切除的可能性,极大地降低了患者的死亡率,从而为中晚期肝癌,尤其是处于巴塞罗那分期B期、C期且未发生全肝转移的肝癌的根治性切除带来了新的希望。因此,ALPPS成为近些年来肝胆外科界的一个热点。然而有些观点认为,ALPPS术式难度大、风险高,且肝脏体积增大不等同肝功能的提高,其实用性还有待评估。也有些研究认为,门静脉结扎能够加快肿瘤生长,而加速肿瘤生长的原因目前尚不清楚。因此ALPPS一期术后残余肝脏组织和肿瘤的病理生理变化,对于指导临床选择最佳的二期手术的时间、减少术后并发症及死亡率尤为重要。
目前在动物建立ALPPS模型已经有许多报道,有助于在临床研究ALPPS术后的病理生理变化。Schlegel等在小鼠建立了第一个动物模型,此后又出现了建立大鼠ALPPS模型的研究,然而这两种模型都有一个共同的问题:肝脏体积的大小造成了进行ALPPS研究具有一定的难度,限制了其广泛应用。Croome等建立的猪ALPPS模型存在着实验成本过高、手术操作难度过大的问题。Mingheng Liao等成功建立了兔ALPPS模型,证实了ALPPS手术在兔实现的可行性,但是以上这些ALPPS动物模型均是在正常的肝脏上进行的,而不是在种植肿瘤的肝脏上,肿瘤的病理生理改变并未考虑在内,对于临床指导工作存在着巨大的缺陷。
发明内容
本发明的目的是针对现有的ALPPS动物模型存在诸多缺点,而提供一种兔VX2肝癌ALPPS手术模型的建立方法,从而降低实验成本,增加动物模型的生存率,增加二期手术的机会,以更全面的、系统的、同步的评价ALPPS一期手术后肿瘤组织及残余肝脏体积的变化,为指导二期手术的开展提供更加科学的依据。
本发明技术解决方案是:一种兔VX2肝癌ALPPS手术模型的建立方法,其特征在于步骤如下:
(1)采用瘤组织块接种法制作VX2兔肝癌模型
将VX2肿瘤组织剪碎成1mm3组织块,于剑突下约 1cm处作一长约 4-5cm的上腹正中切口,逐层切开腹壁,暴露肝脏左外侧叶;于肝叶较厚处,划开一长约3mm的小切口,将瘤块植入,用明胶海绵颗粒填塞肝脏切口,止血,缝合,关腹。
(2)经CT扫描观察,筛选符合尺寸要求的模型,实验组荷瘤兔行ALPPS一期手术
兔肝种植VX2瘤株两周后,肿瘤直径大小可达到约1.0-2.0cm;手术过程:于剑突下偏左注射利多卡因,对切口周围进行麻醉后开腹,切口长约5-7cm,暴露并分离门静脉左支,并用4-0手术缝线结扎,待左肝与右肝之间的缺血线形成,用手术电刀沿着缺血线离断肝实质;用凝胶海绵颗粒填充离断处进行止血,无明显出血后,逐层缝合,关腹,切口消毒。
本发明是首次在兔VX2肝癌上建立ALPPS手术模型。兔VX2肿瘤是肝癌成像研究较为理想的实验模型,与人类肝细胞癌的血供特点和影像学表现非常相似,对临床具有指导作用。而在临床工作中上,ALPPS手术适用于中晚期肝癌患者。因此本发明构建的兔VX2肝癌ALPPS手术模型可以对肿瘤及残余正常肝脏进行整体性的研究。
本发明的优点是:
1、本发明是首次在兔VX2肝癌上建立ALPPS手术模型。以往构建的动物ALPPS手术模型都是基于正常的动物肝脏组织,本项发明所涉及的兔VX2肝癌ALPPS手术模型是基于肝癌进行构建的,符合临床中ALPPS手术在中晚期肝癌患者的应用,极大地弥补了以往ALPPS动物模型对临床指导工作存在的缺陷,并可以为对肿瘤及残余正常肝脏进行深层次研究提供可靠的模型。
2、本手术模型是全程在显微镜下建立的,首先通过寻找Glisson鞘,然后进行进一步的手术处理,极大地提高了手术的精确性以及手术模型的存活率。
3、兔肿瘤模型具有制作简单易行,成本相对较低,实验周期短,成功率高,且VX2肿瘤性质稳定,增殖迅速,短期内即可发生广泛坏死,并形成肝、肺转移,所以VX2肿瘤在大型动物模型的研究上具有很高的应用价值。
4、兔VX2肿瘤是肝癌成像研究较为理想的实验模型,与人类HCC(肝细胞癌)的影像学表现和血供特点具有相似之处,可用作有关肝癌治疗学及肝癌影像学等的相关临床前期研究。
附图说明
图1是兔VX2肝癌模型ALPPS一期手术过程图。
图2 是ALPPS一期术后肝脏和肿瘤大体图。
图3是ALPPS一期术后肝脏和肿瘤体积图。
具体实施方式
一种兔VX2肝癌ALPPS手术模型的建立方法:
步骤一 制作VX2兔肝癌模型
采用肝脏切开癌组织碎块法。首先将VX2瘤株经传代后接种兔后腿外侧肌肉内。约经3周肿瘤生长至直径大小2.0-3.0cm。将兔后腿肌肉内肿瘤取出,剔除坏死组织,选取肿瘤边缘的鱼肉样组织,并用眼科剪将肿瘤组织剪碎成1mm3组织块,用生理盐水反复清洗,于无菌培养皿中备用。将实验兔全麻(麻醉方法:肌肉注射舒泰1ml),呈仰卧位固定于动物手术台上。对兔腹部手术区备皮,消毒手术区皮肤,铺无菌手术洞巾。于剑突下约 1cm处沿腹白线注射利多卡因行局部麻醉,并作一长约 4-5cm 的上腹正中切口,逐层切开腹壁,暴露肝脏左外侧叶;于肝叶较厚处,用眼科镊子划开一长约3mm的小切口,形成一个约3mm×3mm大小的孔道,将3-5个瘤块植入其中,接种后用明胶海绵颗粒填塞肝脏切口,并按压止血,缝合切口,关腹。缝合后于切口处用碘伏消毒,术后3日内常规给予头孢曲松钠40万单位预防感染。每天观察记录实验兔的精神、活动状态,进水量、饮食量和排便量等。
步骤二 兔VX2肝癌模型ALPPS手术
兔肝种植VX2瘤株两周后,经CT扫描观察,肿瘤直径大小可达到约1.0-2.0cm,剔除不符合尺寸要求的模型,实验组荷瘤兔行ALPPS一期手术(即联合门静脉结扎和肝脏离断术)。
60只荷瘤兔随机分成ALPPS组(n =30,肝门静脉左支被结扎,沿着肝左中叶和肝右叶间的缺血线进行肝实质的离断)和Sham组(即假手术组,n =30,未进行门静脉结扎和肝脏离断)。将ALPPS组兔术前禁食12小时,全身麻醉(方法及剂量同前)后,仰卧位固定于兔台上,术区备皮。于剑突下偏左注射利多卡因,对切口周围进行麻醉后开腹,切口长约5-7cm,暴露并分离门静脉左支,并用4-0手术缝线结扎,待左肝与右肝之间的缺血线形成,用电刀沿着缺血线离断肝实质。用凝胶海绵颗粒填充离断处进行止血,无明显出血后,逐层缝合腹壁切口,关闭腹腔,切口处用碘伏消毒。术后3日常规肌肉注射头孢曲松钠40万单位预防感染。Sham组兔子采用ALPPS组兔子相同方式开放,并进行腹腔扰动,关腹后自由生长。每天观察记录实验兔的精神、活动状态,进水量、饮食量、排便量等。
图1中,a肿瘤直径达1.5cm,可行ALPPS手术。b在手术显微镜下找到门静脉。c剥离门静脉左支。d 结扎门静脉左支。e 缺血线形成。f沿着缺血线用电刀进行离断。
图2中,不同时间点离体的兔子肝脏及肿瘤。
图3中,A、肿瘤体积 ,B、残余肝脏体积。
上面描述,只是本发明的具体实施方式,各种举例说明不对本发明的实质内容构成限制。
Claims (1)
1.一种兔VX2肝癌ALPPS手术模型的建立方法,其特征在于步骤如下:(1)采用瘤组织块接种法制作VX2兔肝癌模型将VX2肿瘤组织剪碎成1mm3组织块,于剑突下约 1cm处作一长约4-5cm的上腹正中切口,逐层切开腹壁,暴露肝脏左外侧叶;于肝叶较厚处,划开一长约3mm的小切口,将瘤块植入,用明胶海绵颗粒填塞肝脏切口,止血,缝合,关腹;
(2)经CT扫描观察,筛选符合尺寸要求的模型,实验组荷瘤兔行ALPPS一期手术
兔肝种植VX2瘤株两周后,肿瘤直径大小可达到约1.0-2.0cm;手术过程:于剑突下偏左注射利多卡因,对切口周围进行麻醉后开腹,切口长约5-7cm,暴露并分离门静脉左支,并用4-0手术缝线结扎,待左肝与右肝之间的缺血线形成,用手术电刀沿着缺血线离断肝实质;用凝胶海绵颗粒填充离断处进行止血,无明显出血后,逐层缝合,关腹,切口消毒。
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