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Roux-en-Y胆囊结肠胆汁分流对肝胆系统的影响
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摘要
研究背景
     进行性家族性肝内胆汁淤积(progressive familial intrahepatic cholestasis, PFIC)的非移植外科治疗方式主要包括部分胆汁的外分流术和内分流术。外分流术腹壁永久性胆道外瘘存在,影响儿童正常生活,家属往往难以接受。部分胆汁内分流术虽然避免了胆道外瘘,但是手术操作复杂,部分小肠受损,术后胆汁源性腹泻控制困难。为彻底解决以上问题,我们尝试采用Roux-en-Y胆囊结肠吻合胆汁内分流的新术式。目前有关这一新型改良术式鲜见报道。
     进行性家族性肝内胆汁淤积,以严重的肝内胆汁淤积为主要特征,是一种罕见的常染色体隐性遗传病,发病率1/50000-1/100000,严重危害儿童健康。多数PFIC患儿确诊时已经处于终末期肝病状态。然而由于该症发病机制特殊,很难进行大样本的临床对照研究。为保证实验对象的均一性,动物实验无疑是最佳选择。
     研究目的
     本研究通过建立兔Roux-en-Y胆囊结肠吻合胆汁分流模型,从手术效果、胆汁动力、细菌分析及组织病理等多方面评估该术式的可行性和安全性,为临床应用提供一定的科学理论依据。
     研究方法
     选取24只幼兔随机分为三组,即:(1)正常阴性对照组(n=8),(2)空肠间置部分胆汁内分流组(n=8),(3)Roux-en-Y胆囊结肠吻合组(n=8);分别实施假手术,空肠间置部分胆汁内分流术或Roux-en-Y胆囊结肠吻合胆汁分流术。饲养1年后,行核素肝胆显像评估模型:手术当日及术后1年留取胆汁和血清标本,检测总胆汁酸(total bile acid, TBA)浓度。核素显像完成后,留取肝、胆囊、结肠组织行细菌检测及病理组织学分析。另选取24只幼兔随机分为三组,即:(1)正常对照组(n=8),(2)30厘米长肝支组(n=8)和(3)10厘米短肝支组(n=8);分别实施假手术或Roux-en-Y胆囊结肠吻合术。术后一月行核素肝胆动态显像,获得图像后应用感兴趣区(ROI)技术,由计算机自动绘制时间-放射性曲线。
     实验结果
     观察期间实验兔存活率100%,生长发育正常。经核素肝胆动态显像证实:胆囊结肠吻合组,部分胆汁自胆囊直接分流入结肠,建模成功。胆汁中平均TBA由(801.46±22.54)μmol/L下降至(214.08±12.24)μmol/L,均数下降73%;门静脉血中的TBA含量也呈下降表现,由(85.33±2.08)μmol/L降至(18.85±2.01)均数下降78%。细菌分析:胆囊结肠吻合组,Roux结肠支内细菌总数少于正常对照组,差异有统计学意义。空肠间置内分流组及胆囊结肠吻合组细菌培养阳性率分别为12.5%(4/32),9.375%(3/32);差异不明显。大肠埃希杆菌为最常检出的细菌。病理组织分析发现:肝、胆囊、结肠组织结构基本正常,未见明显畸变结肠隐窝出现。核素动态显像证实:10厘米短支组胆汁排空最快,其20及35分钟胆汁排空率分别为(77.27±6.15)%和(90.39±1.49)%;分流比率最高,达(0.547±0.182),优于长支组(P<0.05)。长支组胆囊排空曲线呈波浪状。
     实验结论
     Roux-en-Y胆囊结肠吻合操作简单,安全可靠,胆汁内分流效果明显;胆囊结肠吻合不会增加细菌移位机率,手术木身与手术后感染无关;观察期内,胆囊结肠分流未诱导兔结肠发生肿瘤性病灶;99mTc-EH1DA核素动态显像技术可以有效评估胆汁分流比率,胆囊排空和整体肝功能变化;在兔的Roux-en-Y胆囊结肠吻合术动物模型中,10厘米短肝支组胆汁内分流效果优于30厘米长肝支组。
Background
     The non-transplanting treatment for progressive familial intrahepatic cholestasis (PFIC) patients included partial external biliary diversion (PEBD) and partial internal biliary drainage (PIBD). PEBD was not readily accepted by all the patients and the families because of the permanent abdominal stoma. A stoma-free partial internal biliary drainage (PIBD) was a complex operation and diarrhea inducing by bile was difficult to treat. Small bowel resection must be carried out. To solve these problems, we have developed a novel approach, Roux-en-Y cholecystocolonic bypass, which partially diverts bile into colon directly without a jejunal conduit. Understanding about its impact on hepatobiliary physiology after this novel operation is limited.
     PFIC was a rare autosomal recessive cholestatic disease with estimated incidence between1/50000and1/100000. Although three types of PFIC had been identified, early diagnosis was still difficult. When diagnosis of PFIC was confirmed, some patients had already progressed to liver fibrosis or cirrhosis. It was difficult to conduct large scale clinical trials or controlled studies. To ensure consistency in the research subjects, an animal model seemed to be the best choice for investigating the effects of cholecystocolonic bypass on hepatobiliary system.
     Objective
     The aim of the present study is to investigate, using a rabbit model, the changes in bile flow, total bile acid (TBA) level, bacterial and histological changes induced by Roux-en-Y cholecystocolonic anastomosis.
     Methods
     24rabbits, randomize divided into3groups:(1)normal control group (n=8),(2) PIBD group (n=8).(3) Roux-en-Y cholecysto-colonic anastomosis group(n=8).1year postoperation, dynamic biligraphy was performed to evaluate the reliability of this animal model. Pre-and post-opertive bile and blood were acquired to examine the change of TBA level. Tissues from the live, gallbladder and colon were also obtained to have pathological test. Bacterial changes also test postperatively. Moreover, another24rabbits were divided into3groups:(1) normal control (n=8),(2)30cm long limb group (n=8) and (3)10cm short limb group (n=8). Group2or3underwent Roux-en-Y cholecysto-colonic anastomosis with a30or10cm length Roux limb, and group1underwent sham operation. One month later, quantitative hepatobiliary scintigraphy was performed to detect the parameters of biliary dynamics using regions of interest (ROI) technology.
     Results
     All animals survived and developed normally during follow-up period. Biligraphy showed that bile was diverted into colon directly after cholecystocolonic anastomosis. Thus, the animal model was successful. TBA level in bile decreased73%from (801.46±22.54)μmol/L to (214.08±12.24)μmol/L. TBA level in portal vein was also decreased78%from (85.33±2.08)μmol/L to (18.85±2.01)μmol/L. After1year, examination of intestinal content showed a significantly decreased number of coliform in the Roux-en-Y cholecysto-colonic group as compared with native control animals. The total positive rate by bacterial culture was12.5%(4/32) and9.375%(3/32) respectively. No difference in the incidence was found among the two groups. E. coli was the most common bacterial cultured. No obvious pathologieal damage to the rabbit's liver, gallbladder and colon after cholecysto-colonic anastomosis was found. Hepatobiliary scintigraphy showed that E20and E35values were (77.27±6.15)%and (90.39±1.49)%respectively in short limb group. That is to say, gallbladder emptying in10cm long limb group was more rapidly than the other two groups. Moreover, the ratio of bile shunt was (0.547±0.182), which was also more than the ratio in the long limb group (P<0.05). The activity-time curve for the GB in the long limb group was like wave.
     Conclusions
     It is feasible and safe to hepatobiliary system after Roux-en-Y cholecysto-colonic anastomosis. Bile bypassed into colon directly, and TBA level in the body was decreased obviously. Roux-en-Y cholecysto-colonic anastomosis itself could not increase the incidence of bacterial translocation, postoperative infections, colorectal cancer induction. Scintigraphy99mTc-EH1DA to assess the ratio of bile diversion, gallbladder motility and overall liver function is a method with satisfactory reproducibility. The effect of bile diversion in10cm short limb rabbit model group is better than the30cm long limb group.
引文
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