兰州大学机构库 >学院待认领
不同方式后处理对大鼠缺血再灌注肾脏的影响
Alternative TitleEffects of postconditionings with different ways on the renal ischemia/reperfusion in rats
姚爱军
Thesis Advisor冷玉芳
2009-05-19
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword缺血/再灌注损伤 缺血后处理 氯胺酮 Fas Caspase-3 大鼠 肾脏 凋亡
Abstract目的:建立成年大鼠肾脏缺血再灌注损伤模型。探讨Fas,Caspase-3在成年大鼠肾脏缺血再灌注损伤肾小管上皮细胞凋亡中的作用和机制;探讨不同方式缺血后处理对成年大鼠肾脏缺血再灌注损伤的影响;探讨两种剂量氯胺酮后处理对成年大鼠肾脏缺血再灌注损伤的影响。 方法:10周龄同批成年健康雄性Wistar大鼠(200±20克)36只,随机分6组,每组6只,即假手术(S)组、缺血再灌注(I/R)组、左下肢缺血后处理(P1)组、肾脏缺血后处理(P2)组、氯胺酮后处理a(K1)组、氯胺酮后处理b(K2)组。S组仅对左肾动脉进行游离,伤口用生理盐水纱布覆盖45min;I/R组采用无创动脉夹夹闭左肾动脉45min后松开,左肾再灌注6h制备I/R模型;P1组在左肾复灌前6min时左股动脉夹闭5min后开放;P2组在左肾缺血45min后灌注10s,停灌10s,反复6次;K1组和K2组在复灌前5min经尾静脉分别给与氯胺酮2mg/Kg和10mg/Kg。检测复灌6h时血清BUN(Cr),Cr(BUN)浓度;光镜下观察肾组织病理改变,原位末端脱氧核苷酸转移酶标记(TUNEL)法检测肾组织中凋亡细胞并计算凋亡指数(AI);免疫组化法检测肾组织Fas,Caspase-3表达,电镜下观察肾单位超微结构改变。使用SPSS10.0进行统计分析,P<0.05为差异有统计学意义。 结果:与S组比较,其他5组大鼠血清BUN、Cr浓度升高(P<0.01),肾组织病理改变明显,肾组织Fas、Caspase-3阳性指数和凋亡指数增加(P<0.01);与I/R组比较,其他4组大鼠血清BUN、Cr浓度降低(P<0.01),肾组织Fas、Caspase-3阳性指数和细胞凋亡指数减少(P<0.01)。与P1组比较,P2组大鼠血清BUN、Cr浓度降低(P<0.05),肾组织Fas、Caspase-3阳性指数和细胞凋亡指数减少(P<0.05)。与K1组比较,K2组大鼠血清BUN、Cr浓度降低(P<0.01),肾组织Fas、Caspase-3阳性指数和细胞凋亡指数减少(P<0.01)。 结论:肾脏缺血再灌注损伤可以启动胞外Fas凋亡通路,通过激活Caspase-3最终导致肾小管上皮细胞凋亡。两种方式缺血后处理可以抑制肾小管上皮细胞凋亡,减轻I/R损伤;肾脏缺血后处理效果更明显。两种剂量氯胺酮后处理都可以抑制肾小管上皮细胞凋亡,减轻I/R损伤,10mg/Kg比2mg/Kg效果更好。 关键词:缺血/再灌注损伤;缺血后处理;氯胺酮;Fas;Caspase-3;大鼠;肾脏;凋亡
Other AbstractObjective: 1. The establishment of an adult rat model of renal ischemia-reperfusion injury. 2. To investigate the role and mechanisms of Fas and Caspase-3 in the (adult rat) renal ischemia-reperfusion injury in apoptosis of renal tubular epithelial cell. 3. To investigate the effects of medical (Ketamine) postconditionings with different doses on rat renal ischemia-reperfusion injury and its molecular mechanism. 4. To investigate the effects of remote (ischemic) postconditionings and the ischemic postconditionings on the renal ischemia-reperfusion injury in rats and its molecular mechanism. Methods: Thirty-six male Wistar rats(10-week-old healthy adult rats) were randomly divided on average into six groups, each with six: sham operation (S) group, ischemia-reperfusion (I/R) group, the left hind limb ischemic postconditioning (P1) group, renal ischemic postconditioning (P2) group, ketamine postconditioning group1 (K1), ketamine postconditioning group 2(K2). In the model of renal ischemia, rats were subjected to unilateral ischemia by clamping of the left renal artery for 45 minutes followed by 6 hours of reperfusion, using a nontraumatic vascular clip, and the right kidneys were removed just after the beginning of the reperfusion of left kidneys. In group S, rats were subjected to the same surgical procedure without clamping the left renal artery. Group P1: One minute before the reperfusion following 45 minutes ischemia in the left kidney, the left femoral arteries have been clipped for 5 minutes; Group P2: these left Kidneys were subjected to six cycles of 10 seconds of reperfusion followed by 10 seconds ischemia immediately after 45 min of ischemia. In group K1 and K2, 2mg/Kg or 10mg/Kg Ketamine were injected via the tail vein 5 min before the reperfusion. The rats were killed after 6 h of reperfusion, and blood samples were collected from right atrium, for measurement of creatinine (Cr) and blood urea nitrogen (BUN). The renal expression of Fas, Caspase-3 stained by immunohistochemistry and the apoptotic cell stained by terminal transferase-mediated dUTP nick-end labeling (TUNEL) were detected through a microscope. Pathological changes of renal tissue were observed under light microscope and electron microscope. The positive cells of Fas, Caspase-3, and apoptosis in tubular epithelial cell were calculated for positive index(PI)and apoptosis index (AI). SPSS10.0 software was used to analyze the data, a=0.05 was considered the level of significance...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223795
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
姚爱军. 不同方式后处理对大鼠缺血再灌注肾脏的影响[D]. 兰州. 兰州大学,2009.
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