肠系膜上静脉血栓形成的实验及临床研究
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摘要
目的:
     总结MVT形成的临床经验,对不同类型MVT的临床特征进行分析,分析影响MVT短期预后的相关因素;评估结扎法建立SMVT大鼠模型的可行性,比较不同部位SMVT对小肠病理生理学的影响;研究不同范围SMVT小肠微循环障碍及体内氧自由基变化规律;研究不同时段使用低分子肝素治疗SMVT的作用机制。
     方法:
     检索2001年1月-2010年12月天津医科大学总医院MVT的临床病例,采集患者的临床数据,包括患病年龄,性别,危险因素史,既往血栓史,临床表现,确诊前症状持续时间,实验室检查,X线及CT检查,手术所见,治疗和临床效果,以及死亡数据,并进行统计学分析;提取可能影响MVT短期预后的相关因素,进行多因素分析。
     以结扎法建立中央型SMVT动物模型。32只大鼠随机分为2组,A组:急性中央型SMVT组,统计大鼠的死亡时间,观察腹腔积液及肠道大体变化,收集肠道标本行组织病理学观察。B组:亚急性中央型SMVT组,术后48h、5d、7d天处死动物,观察腹腔积液及肠道大体变化,收集肠道标本行组织病理学观察。
     以结扎法建立周围型SMVT动物模型。120只大鼠随机分为3组,C组:绞窄性周围型SMVT组;D组:单纯性周围型MVT组;E组:假手术组。分别于术后2h、8h、24h、48h、72h处死动物;观察腹腔大体变化;收集肠道组织及血清学标本;HE染色观察小肠组织病理学变化。
     选取周围型SMVT模型C组、D组、E组各时间点标本;透射电镜观察周围型SMVT肠道微循环障碍及组织损害的超微表现;测定各个时间点血清TXB2、6-Keto-PGF1α、 SOD、MDA的定量指标;统计学分析。
     制作绞窄性周围型SMVT大鼠模型,56只大鼠随机分为3组。按不同干预手段分为低分子肝素治疗组L组、生理盐水对照组N组和假手术组。L组和N组按给药时间不同分为L0h组、L12h组、L24h组和N0h组、N12h组、N24h组6个亚组。72h后观察各组腹腔大体情况;组织病理学观察及病理损伤评分;透射电镜观察微循环的变化;血清TXB2、6-Keto-PGF1α、SOD、 MDA指标检测;统计学分析。
     结果.
     1、MVT病例45例,男性25例,女性20例。入院前症状持续时间为16小时~9个月,平均7.7天。最常见的临床表现为腹痛,其次是腹胀、呕吐、发热等。33例患者接受单纯抗凝、溶栓、祛聚治疗,12例患者接受手术加抗凝、溶栓、祛聚治疗。30天内死亡患者为6例。血液高凝状态在周围型MVT中的比例大于中央型MVT(P=0.031);肝硬化/门静脉高压在中央型MVT所占比例大于周围型MVT(P=0.047)。中央型MVT影像学确诊率高于周围型MVT(P=0.003);周围型MVT发生腹膜炎、接受手术治疗和发生肠坏死的几率高于中央型MVT(P<0.05);中央型MVT发生肠坏死范围更大(P=0.005)。多因素分析表明,最主要的死亡预测因素是年龄≥60岁,相对危险度RR值5.62(95%可信区间);其次是肠切除,相对危险度RR值为3.88(95%可信区间);住院前症状短于3天,相对危险度RR值3.11(95%可信区间)。
     2、A组大鼠术后4h-6h死亡,平均生存时间5.6h。组织病理学检查表现为肠道充血,绒毛顶部上皮细胞无脱落消失,平滑肌无出血及坏死。
     3、B组大鼠良好耐受手术,顺利存活7d。组织病理学检查示肠道淤血性损害,平滑肌层无坏死。术后48h肠系膜上静脉根部周围,胰十二指肠静脉丛周围广泛侧枝循环形成。
     4、C组可以出现典型的透壁性肠梗死过程。2h、8h表现为肠道充血;24h出现粘膜层广泛脱落坏死;48h出现平滑肌层部分坏死;72h出现肠道全层坏死;所有大鼠76-87h内死亡,平均生存时间78.3h。
     5、D组肠道损伤可以恢复。2h、8h表现为肠道充血;24h出现粘膜层部分脱落坏死;48h、72h病理损伤未进一步加重,局限于粘膜层和粘膜下层,平滑肌无出血及坏死;
     6、超微结构观察,C组术后2h血管内大量红细胞淤积;术后8h微血管内可见白细胞贴壁及漏出性出血;术后24h大量微血栓形成;术后48h肠道全层广泛微血栓形成,部分平滑肌结构破坏;术后72h肠道全层出血性坏死。D组术后2h血管内淤血;术后8h肠粘膜上皮细胞线粒体基质变淡;术后24h表现肠粘膜细胞结构破坏,粘膜下层少量微血栓形成;术后48h肠粘膜细胞破坏情况未进行性加重,血管内微血栓形成情况有所减轻;术后72h肠道损害进一步减轻,微血管内少见微血栓形成。
     7、C组各时间点TXB2值高于D组,随时间推移呈升高趋势。D组TXB2在24h时间点达到最大水平,之后有所下降。两组6-Keto-PGF1α值随着时间点的推移逐渐升高,升高幅度不大,各个时间点C组高于D组。两组MDA值逐渐升高趋势,各个时间点C组高于D组。两组SOD值术后一过性升高,C组之后逐渐下降,D组24h后逐渐恢复。
     8、L0h、L12h干预组肠道损害得以转归,无透壁性梗死。血清TXB2、6-Keto-PGF1α、MDA值下降,SOD值上升。L24h组肠道出现透壁性梗死,血清TXB2、6-Keto-PGF1α、MDA值上升,SOD值下降。
     结论:
     1、中央型MVT影像学确诊率高于周围型MVT;周围型MVT发生腹膜炎、接受手术治疗和发生肠坏死的几率高于中央型。
     2、CT诊断早期孤立的小血管静脉血栓在技术是非常困难,连续的观察与渐进式的诊断对于发现MVT非常重要。
     3、确诊前症状短于3天的患者预后更差,有更多的患者需要接受手术治疗,而确诊前症状长于3天的患者较少患者接受手术治疗。
     4、结扎法建立SMVT动物模型适合于SMVT肠道病理生理学研究。
     5、中央型SMVT容易造成循环衰竭,周围型SMVT容易造成肠损害,可以发生透壁性肠梗死。
     6、血性腹水不是SMVT发生肠坏死的标志,血便代表粘膜层和粘膜下层损害。
     7、静脉瘀滞缺氧性损害和肠道微循环障碍是导致SMVT肠损伤的主要始动因素。大量微血栓可以使毛细血管中血液流动更加紊乱,最终结果是肠道的不可逆性坏死。
     8、SMVT导致TXA2/PGI2比例失调,加重肠道微循环障碍。
     9、氧自山基在SMVT发生、发展过程中起着重要的作用,血清MDA和SOD对于SMVT肠损伤严重程度的判断具有重要意义。
     10、LMWH皮下注射可通过降低SMVT大鼠血TXA2、PGI2,改善TXA2/PGI2比例失衡,改善肠道微循环障碍。
     11、LMWH皮下注射能够提高SMVT血液SOD的含量,降低血液的MDA的含量,说明LMWH具有抗氧化、抗炎症作用;
     12、对于绞窄性SMVT,早期抗凝治疗可以显著提高疗效,24h以后抗凝不能逆转肠坏死。
Objective:
     To summarize the experience in the diagnosis and management of SMVT and to assess the factors affecting the short-term outcome.To establish the model of SMVT in rats by ligation method. To observe the pathological physiological variation of the intestinal. To investigate the variation of OFR of Peripheral SMVT. To study the effect of LMWH administration on the intestinal microcirculation of SMVT rats with different time point.
     Methods:
     45patients of MVT in Tianjin Medical University General Hospital were reviewed from2000-2010. The Population characteristics,risk factors, clinical feature, examinations, treatment methods, prognosis were included.
     32rats were divided into group A:acute SMVT of Central type and group B: subacute SMVT of Central type。 The amples of intestinal wall were examined by light electron. The intestinal histopathology score were calculated.
     120rats were divided into group C:strangulated SMVT of peripheral type; group D:simple SMVT of peripheral type; group E:Sham operation group.The models of SMVT were established in rats by ligation method. At2h、8h、24h、48h、72h after operation, amples of intestinal wall and venous blood were obtained.amples of intestinal wall were examined by light and transmission electron microscopy. The TXB2、6-Keto-PGF1α、SOD、MDA were detected from the venous blood.
     56rats were divided into group L: LMWH group; group N:NS group; Sham operation group. strangulated SMVT models were set up. According to different time (Oh,12h,24h),L and N group were each divided into three subgroup randomly. Amples of intestinal wall were examined by light and transmission electron microscopy. The intestinal histopathology score were calculated.The effect was observed by detection the TXB2、6-Keto-PGF1α、SOD、 MDA in the venous blood at72h.
     Results:
     1、45patients were diagnosed MVT.25patients were male,20patients were female,median age52.6years.The main symptoms at the presentation were abdominal pain, abdominal distension,vomiting, bloody stool and fever.Enhancement CT is the most sensitive means.33patients were treated with anticoagulation and12patients were treated with operration.6patients died in30days.
     2、All the rats in group A died from4h to6h and the median survival was5.6h. Histological examinations:the extravasated blood in the each layer was found and the thrombus didn't arise in the mesenteric venous.
     3、The rats in group B tolerated the operation and survival by the7d.The collateral circulation developed around the proximal of the superior mesenteric venous and the pancresticoduodenal veins.The extravasated blood was observed by histological examinations, no necrosis in smooth musle happened.
     4、Histological examinations in group C:the extravasated blood was observed at2h、8h; The serious mucosa necrosis was found at24h;Partly necrosis in smooth musle was found at48h,and the thrombus has arisen in the mesenteric venous;Destruction and necrosis was found in the each layer of intesinal at72h;All the rats died from76h to82h and the median survival was78.3h.
     5、Histological examinations in group D:The extravasated blood was observed at2h、8h; The mild mucosa necrosis was found at24h;The injuries was resricted in the mucosa and submucosa at48h、72h and no necrosis in smooth musle;The thrombus arised in the mesenteric venous at48h.
     6、Ultrastructural observation:Group C:blood stasis was found at2h;The leukocyte sticking along venular wall was found and the blood leaked out from the veins at8h; Large amount of micro thrombus formed at24h; The wide range of micro thrombus formed and the smooth musle cells rupture at48h; Lamellar hemorrhage and necrosis of intesinal could be observed at72h. Group D:Blood stasis was found at2h; Mitochondrion became vacuolization at8h;A few micro thrombus formed at24h; The injuries did not arise at48h; The injuries degraded gradually and few micro thrombus was found at72h.
     7、The results showed:the content of TXB2in group C steped up with the prolongation of the time after operation.and the content was accordingly higher than that in groupB (P<0.05). The content of TXB2in group D steped up before24h,and then degraded gradually.The content of6-Keto-PGF1α、MDA in groupC and group D steped up with the prolongation of the time after operation,and the content of group C was accordingly higher than that in groupB (P<0.05). The content of SOD in groupC signfacanly steped up temporarily after operation,and then degraded gradually. The content of SOD in groupD signfacanly steped up temporarily after operation,and degrade gradually before24h,and then restored after24h.
     7、The intestinal injuries of Group L0h、Group L12h restored. The content of TXB2、6-Keto-PGF1α、MDA degraded and the content of SOD rised.
     Conclusion:
     1、The imaging diagnosis has a higher acuracy in the central SMVT than the peripheral SMVT. The Peripheral SMVT has a greater chance of peritonitis、intestinal necrosis and having surgery.The range of bowel necrosis in the central SMVT is more abroad.
     2、It is difficult of CT to find venule thrombosis and it is important to continuous observation.
     3The patients of symptoms in3days short have a poorer prognosis and is apt to have surgery.
     4、The models of SMVT by ligation method is suitable to the studies of intestinal pathophysiology.
     5、The SMVT of peripheral type is apt to circulatory failure; The SMVT of central type is apt to intestinal injuries.
     6、Hemorrhagic ascites does not mean bowel necrosis;Bloody stool means injureis of the mucosa and submucosa.
     7、The injury of blood stagnation and microcirculatory disturbance are the principal factors of intestinal injuries. Large amount of micro thrombus make the intestinal circulation more serious and lead to bowel necrosis.
     8、SMVT leads to the lack proportion of TXA2/PGI2and make the micro circulation more serious.
     9、 OFR play a vital role on the generation and evolution of SMVT,The level of MDA and SOD is significant judgment criteria to the intestinal injury.
     10、The earlier LMWH administration can reduce the level of TXA2、PGI2and reduce the pathological injury to SMVT rats.
     11、The earlier LMWH administration can raise the level of SOD and reduce the level of MDA.It illusstrates that LMWH can resist to the oxidation and inflammation
     12、Earlier anti-coagulation can improve the results.By the time of24h,it can't reverse the bowel necrosis.
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