【作者】 高志光；

【导师】 李清怀；

【作者基本信息】 河北医科大学 ， 外科学， 2007， 硕士

【摘要】 目的:重症急性胰腺炎(SAP)是临床上比较常见的急腹症之一,其发病急,病势迅猛,多系统器官功能不全综合症(MODS)发生率高,严重时可发生急性肾功能衰竭(ARF)。本文通过动物实验,人为诱导建立大鼠重症急性胰腺炎模型,在零干预的情况下,动态观察肾脏损伤的各项指标,探讨重症急性胰腺炎不同病程时肾损伤的程度,为临床急性胰腺炎患者的诊断及治疗提供参考。方法:取健康SD大鼠96只,雌雄不限,体重300～350克,随机分为对照(SO)组和重症急性胰腺炎(SAP)组,每组实验动物48只。实验前12h开始禁食不禁饮,10%水合氯醛(0.3mg/100g)腹腔注射麻醉,开腹,显露胰胆管,阻断近肝门处胆管后,4号头皮针逆行插入胰胆管并固定,将3.5%牛黄胆酸钠按0.1ml/100g以恒速注入胰胆管,常规关腹,皮下注射生理盐水1ml/100g/h,补充血容量。对照组自胰胆管注入等量生理盐水。每组大鼠于模型制备后3h、6h、9h、12h分为四批,每批12只,经尿集笼留取各时间段尿样,-20℃低温保存,以备测定尿中β2微球蛋白含量;动物心脏采血5ml,随即3000r/min离心分离血清,-80℃低温保存,以备测定血清淀粉酶、BUN浓度;手术切除大鼠胰腺,甲醛固定后沿主胰管方向取胰腺组织一块;取左肾,甲醛固定后经肾门取肾组织一块,常规将胰腺、肾脏标本固定、脱水、浸蜡、包埋、切片。HE染色后光镜观察下观察胰腺形态学变化;肾脏标本一组HE染色后光镜观察下对肾损伤进行组织学评分,一组应用免疫组化测定肾组织中Il-1β的表达率。应用统计学方法对所得结果进行数据分析。结果:光镜观察下:SO组胰腺及肾脏标本均未见明显病理改变;SAP组:胰腺标本可见不同程度腺泡细胞坏死,间质充血、水肿,炎细胞浸润,间质红细胞渗出;肾脏标本可见肾小球萎缩,肾小管扩张,管壁细胞变形,呈扁平状,刷状缘损伤、脱落,细胞浆空泡形成,肾小管腔内有脱落的坏死组织,有时可见管型或碎片形成,间质内可见出血和炎细胞浸润。且随时间延长,胰腺及肾脏的各种病理表现呈加重趋势。血淀粉酶浓度、BUN浓度、肾损伤组织学评分、肾组织中Il-1β的表达率、尿中β2微球蛋白含量的各组数据统计分析结果如下:血淀粉酶浓度:SO组:254.50±76.74;SAP组:4123.08±847.75。以T检验所有数据结果:P<0.05,两组总体均数间差别有显著意义。进一步应用方差分析作各组内比较得出:SO组各时间段间无显著性差异(P>0.05),SAP组内各时间段:3h、6h、9h间均有显著性差异(P<0.05),9h、12h间无显著性差异(P>0.05)。血BUN浓度:SO组:7.44±0.68;SAP组:137.98±89.15。以T检验所有数据结果:P<0.05,两组总体均数间差别有显著意义。进一步应用方差分析作各组内比较得出:SO组各时间段间无显性著差异(P>0.05),SAP组间各时间段间均有显著性差异(P<0.05)。肾损伤组织学评分:SO组:14.13±8.34;SAP组:121.71±43.01。以T检验所有数据结果:P<0.05,两组总体均数间差别有显著意义。进一步应用方差分析作各组内比较得出:SO组各时间段间均无显著性差异(P>0.05);SAP组各时间段间均有显著性差异(P<0.05)。肾组织中Il-1β的表达率:SO组:各时间段均无阳性表达; SAP组:3h组均无阳性表达,6h组有1例阳性表达, 9h组有7例阳性表达,12h组均呈阳性表达。以RXC表进行多个率的比较,SAP组内各时间段间有显著性差异(P<0.05),用确切概率法进一步比较得出:3h组与6h组间无显著性差异(P>0.05),6h、9h、12h间均有显著性差异(P<0.05)。尿中β2微球蛋白含量:SO组:0.101±0.030;SAP组:0.330±0.097。以T检验所有数据结果:P<0.05,两组总体均数间差别有显著意义。进一步应用方差分析作各组内比较得出:SO组各时间段间均无显著性差异(P>0.05),SAP组各时间段间均有显著性差异(P<0.05)。结论:1.重症急性胰腺炎时肾损伤程度随SAP病程延长呈递增趋势;2.血浆BUN浓度对判断SAP时肾损伤程度及预后有重要参考价值. 3. Il-1β在重症急性胰腺炎肾组织中表达显著增加,Il-1β表达增加与SAP肾损伤程度具有密切关系,对于判断SAP肾损伤预后有重要意义;4.尿中β2微球蛋白含量能够较早地反映SAP肾损伤发生和程度。更多还原

【Abstract】 Objective: Severe acute pancreatitis (SAP) is a kind of common surgical acute abdomen disease, acutely morbility, tendency of disease rapidly, high incidence rate of MODS. SAP induces to acute renal failure (ARF) when it is serious enough. This article factitious induced rat’s SAP by animal experiment. Without any of pretreatments on SAP rats and analyzing the change of amylase (AMY)、BUN in serum,β2 microglobulin in urine and the expression ratio of Il-1βin kidney, we investigate the incidence rate and degree of kidney injury during the evolution process and development of SAP. That is to provide a new theory basement for the clinical diagnosis and therapy of SAP.Methods: 96 Sprague-Dawley rat of body weight 300～350g were randomly devided into 2 group, Group A was sham operatio group(n=48). Group B was the severe acute pancreatitis group (n=48). All animal were anaesthesiaed by intraperitoneal injecting 10% chloral hydrate﹙0.3ml/100g﹚in preoperative. Group B was opened abdomon by operating and inserted a scalp acupuncture﹙No 4﹚in pancreas biliary ducts, then pumped 3.5% sodium taurocholate (0.1ml/100g) with constant speed. Group A was acted by injecting 0.9% NS in pancreas biliary ducts. All animals were supplied subcutaneously with 0.9% normal sodium﹙1ml/100g/h﹚after induction of pancreatitis. Animal were killed at 3h, 6h, 9h,12h and accordingly divided to 4 batch (each batch number of group A, B n=12). The serum was centrifugal separated from blood and reserved in -80℃. Urine was collected by urine collection cage and reserved in -20℃.The specimen of pancreas and kidney were exairesised anhydrated、waxed、embedded and cutted sheets. The pathology of kidney and pancreas were observed through light microscope after HE stained. We determined the concentration of AMY and BUN in serum andβ2 microglobulin concentration in urine. Kidney injury was judged by the degree of tubular injury. The expression ratio of Il-1βin kidney was detected by immunohistochemistry. The urineβ2-MG concentration was detected by ELISA. All test results were analysised with the SAS 6.12 system by computer.Results:Observation by light microscope:Samples of kidney and pancreas have no any of pathology change in group SO. But in group SAP, Acinous cell necrosis、stroma hydropsy、inflammation cells infiltrating、RBC exfiltration could be seen in pancreas specimen, nephric globule atrophia、tubular ectasia、border cells cytomorphosis、striated border trauma and amotic、cytolymph vacuolization could be seen in kidney specimen. In some kidney specimen, we find that decididious tissue and necrosis cells、cast and fragements in nephric tubular; haemorrhage and inflammation cells inflatrating in stroma. These pathology changes aggravates with time prolong.The concentration of AMY、BUN in serum andβ2 microglobulin in urine、the expression ratio of Il-1βin kidney tissue and the score of kidney injury in each group were analysised as:The concentration of AMY in serum:Group SO:254.50±76.74;Group SAP:4123.08±847.75;Results analysised by T check discovers:Group SO and group SAP were of statistical significance (p<0.05). Results analysised by ANOVA and S-N-K check discovers:data in group SO has no statistical significance at 3h、6h、9h、12h (p>0.05); data in group SAP has statistical significance at 3h、6h、9h (p<0.05) and it has no statistical significance at 9h、12h(p>0.05).The concentration of BUN in serum:Group SO:7.44±0.68;Group SAP:137.98±89.15;Results analysised by T check discovers : Group SO and group SAP were of statistical significance (p<0.05). Results analysised by ANOVA and S-N-K check discovers : data in group SO has no statistical significance at 3h、6h、9h、12h (p>0.05); data in group SAP has statistical significance at 3h、6h、9h、12h (p<0.05).The score of kidney injury:Group SO:14.13±8.34;Group SAP:121.71±43.01;Results analysised by T check discovers:Group SO and group SAP were of statistical significance (p<0.05). Results analysised by ANOVA and S-N-K check discovers:data in group SO has no statistical significance at 3h、 6h、9h、12h (p>0.05); data in group SAP has statistical significance at 3h、6h、9h、12h (p<0.05).The expression ratio of Il-1βin kidney tissue : The specimen in group SO and group SAP at 3h has no masculine expression; 1 case at 6h and 7 cases at 9h and 12 cases at 12h in group SAP have masculine expression. Results analysised by table R×C discovers : data in group SAP has statistical significance at 3h、6h、9h、12h (p<0.05). Results analysised by exact propability discovers : data in group SAP has no statistical significance at 3h、6h (p>0.05) and has statistical significance at 6h、9h、12h (p<0.05).The concentration ofβ2 microglobulin in urine:Group SO:0.101±0.030;Group SAP:0.330±0.097;Results analysised by T check discovers:group SO and group SAP were of statistical significance (p<0.05). Results analysised by ANOVA and S-N-K check discovers:data in group SO has no statistical significance at 3h、6h、9h、12h (p>0.05); data in group SAP has statistical significance at 3h、6h、9h、12h (p<0.05).Conclusions: 1. Kidney injury ralates to the degree of severe acute pancreatitis. 2. The concentration of BUN in serum has significant meaning in Kidney injury. 3. Il-1βexpression in Kidney tissue closely associated with the digree of Kidney injury in Severe acute pancreatitis, it can be used in judging the results of Kidney injury in Severe acute pancreatitis. 4.The level ofβ2 microglobulin in urine reflects the degree of Kidney injury in Severe acute pancreatitis.更多还原