肝硬化门脉高压大鼠肠道微生态状况的变化及其相关干预的实验研究
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摘要
肝病状态下，尤其在肝硬化门脉高压时，患者常出现如恶心、呕吐、胃纳明显下降等症状，相对而言，肠道菌群营养底物缺乏；肝功能受损其合成能力下降，血浆中的白蛋白水平急聚降低，因而易产生腹水；胆汁分泌不足，肠道内胆盐缺乏；肝脏结构改变，肝硬化门脉高压形成，胃、肠血管动静脉短路开放，血管被动瘀血和主动充血，血管血容量增加，血流瘀滞、组织缺氧，出现门脉高压性胃病及肠病；临床上为防止消化道出血、感染，预防性或治疗性应用抑酸药物如洛赛克和抗生素等。以上因素的存在均可导致肠道微生态失调。表现为专性厌氧菌双歧杆菌、乳酸杆菌显著减少，而革兰阴性杆菌(GNR)显著增多，存在肠道GNR过度性生长、肠管扩张、肠壁变薄等情况。这种失调出现于整个肠道，且肠道微生态失衡程度与肝硬化门脉高压的严重程度相关，并随肝功能出现好转而逐渐恢复。肠道微生态失调，肠道定植抗力下降及肠道屏障功能受损是其肠道内毒素移位、内毒素血症形成的原因之一。业已证明，严重肝病时都存在内毒素血症，其发生率约为58％一100％。内毒素及由内毒素刺激机体产生的TNF-a、IL-1等细胞因子可引起肝细胞凋亡、坏死，加重肝脏原有的损伤。肠道菌群恢复时可以降低门静脉内毒素，但是大鼠肠道内毒素的下降要晚于肠道菌群的恢复。随着肝硬化门脉高压病情的恶化，可出现各种并发症，其中与肠道微生态有密切关系的有继发性细菌感染、内毒素血症、上消化道出血等。因此，在治疗肝硬化门脉高压症时，尚需从肠道微生态的角度加以对其并发症的防治。消化道出血历来是外科研究和治疗的重点。门脉高压性胃病、门脉高压性肠病等所致的消化道出血、贫血、消化吸收不良、菌群失调和感染等病症相对缺乏防、治经验。本研究：
目的：
1、构建稳定、可靠的肝硬化门脉高压大鼠动物模型，研究该模型肝脏及肠道病变情况。
2、研究肝硬化门脉高压大鼠菌群移位、肠道微生态、肠黏膜屏障功能及内毒素血症等改变，并探讨TNF-α、对Occludin蛋白以及肠黏膜屏障功能的影响，在肠源性内毒素血症发病机制中的作用。
3、研究微生态制剂-乳果糖在调节门脉高压大鼠肠道菌群失调、抑制菌群移位、改善内毒素血症进而改善肝功能的作用，探讨其在肝硬化门脉高压治疗中的作用机制。
4、研究微生态制剂-乳果糖对行脾切除+选择性断流术后门脉高压大鼠肠道微生态状况的影响，探讨其在门脉高压症外科治疗中的意义。
方法：
1、肝硬化门脉高压大鼠动物模型建立：按0．4 ml／100g剂量予以背部皮下注射50％CCL_4橄榄油溶液，每周2次，持续4周，第5周开始改以0．5ml／100g标准背部皮下注射，每周2次，持续4周，模型制作共10周。行上腹部CT、MR扫描，观察肝硬化、脾肿大、腹水等情况；以HE染色、Masson三色染色、透射电镜观察大鼠肝脏及肠组织病变情况；测量门静脉直径和门静脉压力以了解该模型血液动力学变化情况。
2、电镜观察肠上皮细胞紧密连接超微结构和绒毛病变；检测细菌易位率；分析肠道菌群构成情况并作菌种DNA指纹图谱分析；做肝功能测定；采用改良的偶氮基质显色法行内毒素测定；采用免疫组织化学方法和Western blot法分别测定其末段回肠和结肠的肿瘤坏死因子(tumor necrosis factorα，TNF-α)及紧密连接蛋白(Occludin)表达水平。
3、对门脉高压模型大鼠进行乳果糖干预治疗即：每日灌服乳果糖(67％)5ml／kg，每日2次，直至大鼠排稀便，研究其肠黏膜通透性及肠微生态改变，检测细菌移位、肝功能、内毒素血症情况。
4、将门脉高压大鼠行“脾切除加选择性贲门周围血管离断术”后，应用乳果糖干预治疗，研究其肠黏膜通透性及肠微生态改变，检测细菌移位、肝功能、内毒素血症情况。
结果：
1、复合法四氯化碳大鼠门脉高压模型，死亡率28％；四氯化碳用药10周后见大鼠体形消瘦，CT检查示肝脏体积缩小，脾脏略有增大，伴有肝周少量腹水；病理切片见肝内假小叶广泛形成，假小叶内可见纤维再分割现象，同时伴有明显的肝细胞再生结节；开腹穿刺测压结果：对照组(7．62±2．02)mmHg、实验组(15．76±2．68)mmHg，两组比较具有显著性差异(P＜0．05)。表明存在肝硬化门静脉高压表现。
2、门脉高压组细菌易位发生率较正常对照组明显增高(P＜0．05)，乳杆菌、双歧杆菌的数量则显著减少(P＜0．01)，而肠杆菌、肠球菌数量则明显增多(P＜0．01)；门脉高压组肝功能各项(AST、ALT、TBLL、ALB)叫正常组均明显恶化(P＜0．01)，其血清内毒素水平(0．44±0．07Eu／ml)亦明显高于正常组之(0．20±0．08Eu／ml)(P＜0．01)；回、结肠的紧密连接蛋白Occludin及TNF-α表达，两组比较具有显著性差异(P＜0．01)。
3、应用乳果糖干预后，门脉高压治疗对照组(A组)细菌易位发生率较乳果糖组(B组)和正常对照(C组)均明显增高(P＜0．05)；B组及C组乳杆菌、双歧杆菌的数量均较A组增多(P＜0．01)，而肠杆菌、肠球菌数量则显著降低(P＜0．01)；A组细菌DNA指纹图谱条带明显减少，且出现异常条带，B组肠道内优势菌群的基因条带与C组具有高度一致性；与B、C组比较，A组肝功能各项(AST、ALT、TBLL、ALB)均明显恶化(P＜0．01)；A组血清内毒素水平(0．44±0．07Eu／m1)亦明显高于B、C组之(0．33±0．06Eu／m1)、(0．20±0．08Eu／m1)(P＜0．01)；回、结肠的紧密连接蛋白及TNF-α表达，A组与B、C组比较，具有显著性差异(P＜0．01)。
4、行“脾切除加选择性贲门周围血管离断术”后门脉高压大鼠，应用乳果糖干预治疗。术后治疗对照组(PC组)细菌易位发生率较术后乳果糖组(PB组)和假手术组(PD组)均明显增高(P＜0．05)；PB组及PD组乳杆菌、双歧杆菌的数量均较PC组增多(P＜0．01)，而肠杆菌、肠球菌数量则显著降低(P＜0．01)；PC组细菌DNA指纹图谱条带明显减少，B组肠道内优势菌群的基因条带与术前治疗组(A组)、PD组无明显差异；与PB、PD组比较，PC组肝功能各项(AST、ALT、TBLL)均明显升高(P＜0．01)，而血清白蛋白(ALB)则显著降低(P＜0．01)；PC组血清内毒素水平(0．51±0．08Eu／ml)亦明显高于PB、PD组之(0．31±0．09Eu／ml)、(0．34±0．09Eu／ml)(P＜0．01)；回、结肠的紧密连接蛋白及TNF-α表达，PC组与PB、PD组比较，具有显著性差异(P＜0．01)。
结论：
我们应用四氯化碳橄榄油复合法所构建的动物模型其组织学改变与人类肝硬变相似，门静脉压力显著增高并产生一定程度的门体交通支分流，是研究门脉高压症的理想模型。
门脉高压大鼠随着病变的进展，存在着明显的肠黏膜屏障功能障碍，通透性升高，出现肠道微生态紊乱，菌群失调，菌群移位和肠源性内毒素血症，同时肝功能损害加剧。
微生态制剂乳果糖可以明显改善门脉高压大鼠肠道的微生态，可促进有益菌的生长从而提高定植抗力，并籍此抑制潜在致病菌生长及其有害代谢物的产生，减少肠源性内毒素的产生，促进肝功能的恢复。乳果糖的这种作用不仅适用于未接受手术治疗的门脉高压大鼠，在接受“脾切除加选择性贲门周围血管离断术”的门脉高压大鼠中同样作用显著，这为临床提高门静脉高压症患者手术疗效，缓解近、远期并发症、提高患者生存质量，提供了一定的理论依据和积累了经验。
OBJECTIVE
1. Establish a steady、credible rat model of cirrhosis portal hypertension. Study its pathological changes of live and intestine.
2. To study the bacterial translocation, intestinal microecology, gut barrier and endotoxemia. Explore the effects of TNF-αon the Occludin protein and the intestinal barrier and its role in the pathogenesy of endotoxemia origin from intestine.
3. To Research on the effects of microecological preparation-lactulose on conditioning the imbalance of intestinal flora of rats with portal hypertension, inhibiting bacterial translocation, attenuating endotoxemia and improving liver function, explore its role of mechanisms in treatment of portal hypertension.
4. To Research on the effects of microecological preparation-lactulose on the intestinal microecology of rats with portal hypertension that have recived splenectomy and devascularization, to explore its significance in surgical treatment of portal hypertension.
METHODS
1. Models of portal hypertension were established as follows: 50% CC14 olive oil was administered parenterally on the back of rats at a dose of 0.4 mL/100 g body weight twice a week for consecutive 4 weeks. On the 5th week, the dose was changed to 0.5 mL/100 g and the medication was given twice a week for 4 weeks. The preparation of portal hypertension model lasted 10 weeks. The rats received upper abdominal CT, MR scanning. Observed the conditions of cirrhosis, splenomegaly, ascites ect. Study the pathological changes of live and intestine tissues by using HE staining, Masson staining and Transmission Electron Microscope, respectively. The diameter of portal vein and portal venous pressure was used to observe the change of flow dynamics.
2. Ultra-structure of intertinal epithelial cells and villus pathological changes were observed under electron microscope., the bacterial translocation rate was calculated, The contents of cecum were examined for the growth of anaerobic bacteria. Randomly amplified polymorphic DNA (RAPD) technique was used for the analysis of DNA fingerprint. Blood was sampled from abdominal aorta for the measurement of liver function. Modified diazo-coupling method was employed for the detection of endotoxin. The expressions of TNF-αand Occludin were determined in the tissues of terminal ileum and colon by using immunochemistry and Western blot respectively.
3. The treatment with lactulose was given to rats with portal hypertension: the rats were gavaged with 5 ml/kg lactulose twice daily when rats began to defecate loose stools, then carried out our research as step 2.
4. The portal hypertension rats were given splenectomy and Periesophagogastric devascularization, then carried out our research as step 3.
RESULTS
1. The mortality rate of rat's model of portal hypertension is 28%. 10 weeks after the rats were subcutaneously injected of 50% CCL_4 olive oil on the back, weight loss was seen, CT revealed that the liver volume was reduced and spleen enlarged, which was accompanied with a small quantity of ascites in the perihepatic tissues. Pathological examination revealed plenty of pseudolobules containing fibrous septa and regenerative nodules of liver cells. The intra-abdominal pressure measurement showed that the pressure of the experimental group was significantly higher than that of the control group (15.76±2.68 mmHg vs. 7.62±2.02 mmHg, P<0.05)
2. The bacterial translocation rate was increased in portal hypertension group when compared with that in normal group (P<0.05). The number of lactobacillus and bacillus bifidus was declined and that of enterobacteria and enterococcus elevated (P<0.01). Compared with that in normal group, the liver function profile was significantly worsed in portal hypertension group (P<0.01). And the endotoxin level (0.44±0.07Eu/ml) was much higher in portal hypertension group than normals (0.20±0.08Eu/ml) (P<0.01). There were significant differences in the expressions of TNF-αand Occludin between the two groups (P<0.01).
3. After treated with lactulose, the bacterial translocation rate was increased in portal hypertension treatment control group (group A) when compared with that in lactulose group (B) and normal control group (C) (P<0.05). The number of lactobacillus and bacillus bifidus was declined and that of enterobacteria and enterococcus elevated in group A as compared with those in groups B and C (P<0.01). Less bands and abnormal bands were found on the DNA fingerprint map of group A. Gene bands of dominant bacterial flora in group B were akin to those in group C. Compared with that in groups B and C, the liver function profile was significantly worsed in group A. And the endotoxin level was much higher in group A than in groups B and C. There were significant differences in the expressions of TNF-αand Occludin between group A and groups B, C (P<0.01).
4. The portal hypertension rats that had received "splenectomy plus pericardial devascularization" were gavaged with lactulose. The bacterial translocation rate was increased in treatment control group postoperation (group PC) when compared with that in lactulose group postoperation (group PB) and sham operation group (group PD) (P<0.05). The number of lactobacillus and bacillus bifidus was elevated and that of enterobacteria and enterococcus declined in groups PB and PD as compared with those in group PC (P<0.01). Less bands were found on the DNA fingerprint map of group PC. Compared with that in groups PB and PD, the liver function profile was significantly worsed in group PC. And the endotoxin level was much higher in group PC than in groups PB and PD. There were significant differences in the expressions of TNF-αand Occludin between group PC and groups PB, PD (P<0.01).
CONCLUSION
The model of portal hypertension establish by subcutaneous injection of 50% CCL_4 Olive oil on the back of rats has a similar histology change to human cirrhosis. The portal venous pressure raised significantly and certain level's of port-body branches' shunts occurred, It is a appropriate model to study portal hypertension.
The development of portal hypertension is accompanied by the damage of intestinal barrier. The raise of intestinal permeability is the main reason for intestinal endotoxemia. The disturbanc of intestinal microecology, bacterial translocation, dysbacteria and intestinal endotoximia occurred, meanwhile liver function got worse.
The treatment with lactulose can successfully improve the intestinal environment, enhance the ability of intestine to ward off microbial colonization, reduce the bacterial translocation, and attenuate endotoxemia, thereby improving the liver function. It's these kinds of functions refer to not only portal hypertension rats that have not received operation but also those that have undergone the operation. This also provided a theoretical basis and accumulated experience for improving surgical treatment and quality of life in patients with portal hypertension and alleviating the short-term and long-term complications in clinic.
目的：
1、构建稳定、可靠的肝硬化门脉高压大鼠动物模型，研究该模型肝脏及肠道病变情况。
2、研究肝硬化门脉高压大鼠菌群移位、肠道微生态、肠黏膜屏障功能及内毒素血症等改变，并探讨TNF-α、对Occludin蛋白以及肠黏膜屏障功能的影响，在肠源性内毒素血症发病机制中的作用。
3、研究微生态制剂-乳果糖在调节门脉高压大鼠肠道菌群失调、抑制菌群移位、改善内毒素血症进而改善肝功能的作用，探讨其在肝硬化门脉高压治疗中的作用机制。
4、研究微生态制剂-乳果糖对行脾切除+选择性断流术后门脉高压大鼠肠道微生态状况的影响，探讨其在门脉高压症外科治疗中的意义。
方法：
1、肝硬化门脉高压大鼠动物模型建立：按0．4 ml／100g剂量予以背部皮下注射50％CCL_4橄榄油溶液，每周2次，持续4周，第5周开始改以0．5ml／100g标准背部皮下注射，每周2次，持续4周，模型制作共10周。行上腹部CT、MR扫描，观察肝硬化、脾肿大、腹水等情况；以HE染色、Masson三色染色、透射电镜观察大鼠肝脏及肠组织病变情况；测量门静脉直径和门静脉压力以了解该模型血液动力学变化情况。
2、电镜观察肠上皮细胞紧密连接超微结构和绒毛病变；检测细菌易位率；分析肠道菌群构成情况并作菌种DNA指纹图谱分析；做肝功能测定；采用改良的偶氮基质显色法行内毒素测定；采用免疫组织化学方法和Western blot法分别测定其末段回肠和结肠的肿瘤坏死因子(tumor necrosis factorα，TNF-α)及紧密连接蛋白(Occludin)表达水平。
3、对门脉高压模型大鼠进行乳果糖干预治疗即：每日灌服乳果糖(67％)5ml／kg，每日2次，直至大鼠排稀便，研究其肠黏膜通透性及肠微生态改变，检测细菌移位、肝功能、内毒素血症情况。
4、将门脉高压大鼠行“脾切除加选择性贲门周围血管离断术”后，应用乳果糖干预治疗，研究其肠黏膜通透性及肠微生态改变，检测细菌移位、肝功能、内毒素血症情况。
结果：
1、复合法四氯化碳大鼠门脉高压模型，死亡率28％；四氯化碳用药10周后见大鼠体形消瘦，CT检查示肝脏体积缩小，脾脏略有增大，伴有肝周少量腹水；病理切片见肝内假小叶广泛形成，假小叶内可见纤维再分割现象，同时伴有明显的肝细胞再生结节；开腹穿刺测压结果：对照组(7．62±2．02)mmHg、实验组(15．76±2．68)mmHg，两组比较具有显著性差异(P＜0．05)。表明存在肝硬化门静脉高压表现。
2、门脉高压组细菌易位发生率较正常对照组明显增高(P＜0．05)，乳杆菌、双歧杆菌的数量则显著减少(P＜0．01)，而肠杆菌、肠球菌数量则明显增多(P＜0．01)；门脉高压组肝功能各项(AST、ALT、TBLL、ALB)叫正常组均明显恶化(P＜0．01)，其血清内毒素水平(0．44±0．07Eu／ml)亦明显高于正常组之(0．20±0．08Eu／ml)(P＜0．01)；回、结肠的紧密连接蛋白Occludin及TNF-α表达，两组比较具有显著性差异(P＜0．01)。
3、应用乳果糖干预后，门脉高压治疗对照组(A组)细菌易位发生率较乳果糖组(B组)和正常对照(C组)均明显增高(P＜0．05)；B组及C组乳杆菌、双歧杆菌的数量均较A组增多(P＜0．01)，而肠杆菌、肠球菌数量则显著降低(P＜0．01)；A组细菌DNA指纹图谱条带明显减少，且出现异常条带，B组肠道内优势菌群的基因条带与C组具有高度一致性；与B、C组比较，A组肝功能各项(AST、ALT、TBLL、ALB)均明显恶化(P＜0．01)；A组血清内毒素水平(0．44±0．07Eu／m1)亦明显高于B、C组之(0．33±0．06Eu／m1)、(0．20±0．08Eu／m1)(P＜0．01)；回、结肠的紧密连接蛋白及TNF-α表达，A组与B、C组比较，具有显著性差异(P＜0．01)。
4、行“脾切除加选择性贲门周围血管离断术”后门脉高压大鼠，应用乳果糖干预治疗。术后治疗对照组(PC组)细菌易位发生率较术后乳果糖组(PB组)和假手术组(PD组)均明显增高(P＜0．05)；PB组及PD组乳杆菌、双歧杆菌的数量均较PC组增多(P＜0．01)，而肠杆菌、肠球菌数量则显著降低(P＜0．01)；PC组细菌DNA指纹图谱条带明显减少，B组肠道内优势菌群的基因条带与术前治疗组(A组)、PD组无明显差异；与PB、PD组比较，PC组肝功能各项(AST、ALT、TBLL)均明显升高(P＜0．01)，而血清白蛋白(ALB)则显著降低(P＜0．01)；PC组血清内毒素水平(0．51±0．08Eu／ml)亦明显高于PB、PD组之(0．31±0．09Eu／ml)、(0．34±0．09Eu／ml)(P＜0．01)；回、结肠的紧密连接蛋白及TNF-α表达，PC组与PB、PD组比较，具有显著性差异(P＜0．01)。
结论：
我们应用四氯化碳橄榄油复合法所构建的动物模型其组织学改变与人类肝硬变相似，门静脉压力显著增高并产生一定程度的门体交通支分流，是研究门脉高压症的理想模型。
门脉高压大鼠随着病变的进展，存在着明显的肠黏膜屏障功能障碍，通透性升高，出现肠道微生态紊乱，菌群失调，菌群移位和肠源性内毒素血症，同时肝功能损害加剧。
微生态制剂乳果糖可以明显改善门脉高压大鼠肠道的微生态，可促进有益菌的生长从而提高定植抗力，并籍此抑制潜在致病菌生长及其有害代谢物的产生，减少肠源性内毒素的产生，促进肝功能的恢复。乳果糖的这种作用不仅适用于未接受手术治疗的门脉高压大鼠，在接受“脾切除加选择性贲门周围血管离断术”的门脉高压大鼠中同样作用显著，这为临床提高门静脉高压症患者手术疗效，缓解近、远期并发症、提高患者生存质量，提供了一定的理论依据和积累了经验。
OBJECTIVE
1. Establish a steady、credible rat model of cirrhosis portal hypertension. Study its pathological changes of live and intestine.
2. To study the bacterial translocation, intestinal microecology, gut barrier and endotoxemia. Explore the effects of TNF-αon the Occludin protein and the intestinal barrier and its role in the pathogenesy of endotoxemia origin from intestine.
3. To Research on the effects of microecological preparation-lactulose on conditioning the imbalance of intestinal flora of rats with portal hypertension, inhibiting bacterial translocation, attenuating endotoxemia and improving liver function, explore its role of mechanisms in treatment of portal hypertension.
4. To Research on the effects of microecological preparation-lactulose on the intestinal microecology of rats with portal hypertension that have recived splenectomy and devascularization, to explore its significance in surgical treatment of portal hypertension.
METHODS
1. Models of portal hypertension were established as follows: 50% CC14 olive oil was administered parenterally on the back of rats at a dose of 0.4 mL/100 g body weight twice a week for consecutive 4 weeks. On the 5th week, the dose was changed to 0.5 mL/100 g and the medication was given twice a week for 4 weeks. The preparation of portal hypertension model lasted 10 weeks. The rats received upper abdominal CT, MR scanning. Observed the conditions of cirrhosis, splenomegaly, ascites ect. Study the pathological changes of live and intestine tissues by using HE staining, Masson staining and Transmission Electron Microscope, respectively. The diameter of portal vein and portal venous pressure was used to observe the change of flow dynamics.
2. Ultra-structure of intertinal epithelial cells and villus pathological changes were observed under electron microscope., the bacterial translocation rate was calculated, The contents of cecum were examined for the growth of anaerobic bacteria. Randomly amplified polymorphic DNA (RAPD) technique was used for the analysis of DNA fingerprint. Blood was sampled from abdominal aorta for the measurement of liver function. Modified diazo-coupling method was employed for the detection of endotoxin. The expressions of TNF-αand Occludin were determined in the tissues of terminal ileum and colon by using immunochemistry and Western blot respectively.
3. The treatment with lactulose was given to rats with portal hypertension: the rats were gavaged with 5 ml/kg lactulose twice daily when rats began to defecate loose stools, then carried out our research as step 2.
4. The portal hypertension rats were given splenectomy and Periesophagogastric devascularization, then carried out our research as step 3.
RESULTS
1. The mortality rate of rat's model of portal hypertension is 28%. 10 weeks after the rats were subcutaneously injected of 50% CCL_4 olive oil on the back, weight loss was seen, CT revealed that the liver volume was reduced and spleen enlarged, which was accompanied with a small quantity of ascites in the perihepatic tissues. Pathological examination revealed plenty of pseudolobules containing fibrous septa and regenerative nodules of liver cells. The intra-abdominal pressure measurement showed that the pressure of the experimental group was significantly higher than that of the control group (15.76±2.68 mmHg vs. 7.62±2.02 mmHg, P<0.05)
2. The bacterial translocation rate was increased in portal hypertension group when compared with that in normal group (P<0.05). The number of lactobacillus and bacillus bifidus was declined and that of enterobacteria and enterococcus elevated (P<0.01). Compared with that in normal group, the liver function profile was significantly worsed in portal hypertension group (P<0.01). And the endotoxin level (0.44±0.07Eu/ml) was much higher in portal hypertension group than normals (0.20±0.08Eu/ml) (P<0.01). There were significant differences in the expressions of TNF-αand Occludin between the two groups (P<0.01).
3. After treated with lactulose, the bacterial translocation rate was increased in portal hypertension treatment control group (group A) when compared with that in lactulose group (B) and normal control group (C) (P<0.05). The number of lactobacillus and bacillus bifidus was declined and that of enterobacteria and enterococcus elevated in group A as compared with those in groups B and C (P<0.01). Less bands and abnormal bands were found on the DNA fingerprint map of group A. Gene bands of dominant bacterial flora in group B were akin to those in group C. Compared with that in groups B and C, the liver function profile was significantly worsed in group A. And the endotoxin level was much higher in group A than in groups B and C. There were significant differences in the expressions of TNF-αand Occludin between group A and groups B, C (P<0.01).
4. The portal hypertension rats that had received "splenectomy plus pericardial devascularization" were gavaged with lactulose. The bacterial translocation rate was increased in treatment control group postoperation (group PC) when compared with that in lactulose group postoperation (group PB) and sham operation group (group PD) (P<0.05). The number of lactobacillus and bacillus bifidus was elevated and that of enterobacteria and enterococcus declined in groups PB and PD as compared with those in group PC (P<0.01). Less bands were found on the DNA fingerprint map of group PC. Compared with that in groups PB and PD, the liver function profile was significantly worsed in group PC. And the endotoxin level was much higher in group PC than in groups PB and PD. There were significant differences in the expressions of TNF-αand Occludin between group PC and groups PB, PD (P<0.01).
CONCLUSION
The model of portal hypertension establish by subcutaneous injection of 50% CCL_4 Olive oil on the back of rats has a similar histology change to human cirrhosis. The portal venous pressure raised significantly and certain level's of port-body branches' shunts occurred, It is a appropriate model to study portal hypertension.
The development of portal hypertension is accompanied by the damage of intestinal barrier. The raise of intestinal permeability is the main reason for intestinal endotoxemia. The disturbanc of intestinal microecology, bacterial translocation, dysbacteria and intestinal endotoximia occurred, meanwhile liver function got worse.
The treatment with lactulose can successfully improve the intestinal environment, enhance the ability of intestine to ward off microbial colonization, reduce the bacterial translocation, and attenuate endotoxemia, thereby improving the liver function. It's these kinds of functions refer to not only portal hypertension rats that have not received operation but also those that have undergone the operation. This also provided a theoretical basis and accumulated experience for improving surgical treatment and quality of life in patients with portal hypertension and alleviating the short-term and long-term complications in clinic.
引文
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