【作者】 张宁；

【导师】 刘勇；

【作者基本信息】 中国医科大学 ， 临床检验诊断学， 2010， 硕士

【摘要】 目的研究不同浓度内毒素对肾脏的早期(内毒素作用后的前5小时)影响,以试图找出在此段时间内不同剂量LPS作用下肾脏损伤的变化规律,从而为临床内毒素血症患者的肾脏病变及早治疗干预提供参考依据。方法将48只大耳白兔随机分为对照组和LPS (E.Coli O55B5)的高(400μg／kg)、中(200μg／kg)、低(100μg／kg)三个剂量组。对照组3只,其余每组15只(每组有5个时间点,每个时间点3只,均为随机分配)。依据体重沿耳缘静脉注入相应量的溶于5ml生理盐水的LPS,观察注射后第1、2、3、4、5小时,分别对应为T1、T2、T3、T4、T5这5个时间点的肾脏变化情况。每个时间点取肾脏组织进行病理检查,并利用免疫组化法检测肾脏细胞凋亡状况、生化检测血肌酐和血尿素、电泳法分析检测尿液中的蛋白质。结果对照组血清尿素从T1至T5分别为：6.0±0.9,6.1±1.0,6.1±0.9,6.1±1.0,6.2±1.1mmol／L,6.2±1.1mmol／L,对照组血肌酐从T1至T5分别为：59±13,59±10,59±12,58±10,59±12μmol／L；低剂量组血尿素从T1至T5分别为：6.4±1.2,7.7±1.8,8.2±0.9,9.1±1.2,10.7±1.1mmol／L,与对照组相比,T4出现显著性差异(P<0.05),血肌酐从T1至T5分别为：55±15,64±19,81±17,111±15,121±17μmol／L,与对照组相比,T4出现显著性差异(P<0.05)；中剂量组血尿素从T1至T5分别为：7.0±1.8,8.4±1.7,9.2±1.3,9.6±1.4,11.8±1.8mmol／L,与对照组相比,T3出现显著性差异(P<0.05),血肌酐从T1至T5分别为：71±7,77±9,97±8,122±14,152±11μmol／L,与对照组相比,T3出现显著性差异(P<0.05)；高剂量组血尿素从T1至T5分别为：9.4±1.2,10.8±1.5,12.1±1.9,12.7±1.8,13.8±1.2mmol／L,与对照组相比,T1出现显著性差异(P<0.05),血肌酐从T1至T5分别为：84±9,101±6,126±14,158±16,206±23μmol／L,与对照组相比,T1出现显著性差异(P<0.05)。肾脏病理出现损伤以及免疫组化出现肾脏细胞凋亡均见于低剂量组的T5,中剂量组的T4和高剂量组的T3；尿蛋白电泳出现管性蛋白尿结果的各组时间点与上述一致。结论本实验低、中、高三个LPS剂量组在出现肾脏病理损伤前均出现氮质血症,随着LPS作用剂量的增加和作用时间的延长,兔血清肌酐和尿素水平逐渐增高,并出现管性蛋白尿,呈现出肾脏损伤与LPS的剂量、时间依赖性。更多还原

【Abstract】 ObjectiveTo find a general rule of renal function damage in different level of endotoxemia model rabbit during the first 5 hours and further provide a reference for clinical treatment of endotoxemia.MethodsForty-eight big-eared rabbits were randomly divided into 4 groups:normal control group, low-dose of LPS group(100μg/kg), middle-dose of LPS group(200μg/kg) and high-dose of LPS group(400μg/kg). The normal control group included 3 rabbits, and the other groups were 15 of each (3 rabbits for every time point). According to the weight and group, each rabbits were injected with different dose of LPS dissolved in 5ml NS. The normal control group were injected with 5ml NS without LPS in ten minutes. The observation was started after the injection was completed. There were 5 time points as follows:1st hour(T1),2nd hour(T2),3rd hour(T3),4th hour(T4) and 5th hour(T5). Each time point, every group had 3 rabbits to be dealt with anaesthetizing and then the blood sample were drawn to measure Urea and Scr, urine were collected from the bladder for electropHoresis, and then the kidneys were dissected for pathology and immunol histochemistry assay.ResultsThe normal control group’s Urea from T1 to T5 were as follows:6.0±0.9,6.1±1.0, 6.1±0.9,6.1±1.0,6.2±1.1mmol/L, Scr from T1 to T5 were as follows:59±13,59±10, 59±12,58±10,59±12μmol/L. While the low-dose of LPS group’s Urea (from T1 to T5 were as follows:6.4±1.2,7.7±1.8,8.2±0.9,9.1±1.2,10.7±1.1mmol/L)was significantly increased at T4(P<0.05), Scr(from T1 to T5 were as follows:55±15,64±19,81±17, 111±15,121±17μmol/L)was significantly increased at T4(P<0.05); middle-dose of LPS group’s Urea (from T1 to T5 were as follows:7.0±1.8,8.4±1.7,9.2±1.3,9.6±1.4, 11.8±1.8mmol/L) was significantly increased at T3(P<0.05), Scr(from T1 to T5 were as follows:71±7,77±9,97±8,122±14,152±11μmol/L)was significantly increased at T3(P<0.05);high-dose of LPS group’s Urea (from T1 to T5 were as follows:9.4±1.2, 10.8±1.5,12.1±1.9,12.7±1.8,13.8±1.2mmol/L)was significantly increased at T1(P<0.05), Scr(from T1 to T5 were as follows:84±9,101±6,126±14,158±16, 206±23μmol/L)was significantly increased at T1(P<0.05). The significantly different result of the urine electropHoresis was demonstrated at T5 in low-dose of LPS group, T4 in middle-dose of LPS group, T3 in high-dose of LPS group, the similar results were also observed in the tests of pathology and immunol histochemistry assay.ConclusionAll of the three treatment groups appeared azotemia before kidney pathological injury were observed. Urea and Scr were significantly increased, and tubular proteinuria were observed. Kidney injury induced by endotoxin became worsened on a dose-effect and time-effect dependent style.更多还原