【作者】 王俊；

【导师】 严海东；

【作者基本信息】 同济大学 ， 内科学， 2007， 硕士

【摘要】 目的研究不同透析膜对维持性血液透析(maintenance hemodialysis MHD)患者血浆及外周血单个核细胞(peripherial blood mononuclear cells PBMC)合成、分泌白细胞介素13(interleukin 13 IL-13)和白细胞介素18(interleukin 18 IL-18)水平以及氧化应激的影响;并判断IL-13、IL-18能否作为评价透析膜生物相容性的指标。方法将20名MHD患者随机分为纤维素生物膜组(650组)和聚砜膜组(F6组),每组各10例。应用酶联免疫吸附(enzyme linked immunosorbent assay ELISA)法测定MHD患者透析前、后血浆中以及PBMC经或未经刺激培养上清中IL-13、IL-18水平。采用半定量逆转录多聚酶链反应(Reverse Transcription PolymeraseChain Reaction RT-PCR)技术检测PBMC中IL-13、IL-18 mRNA的表达。同时选取30名MHD患者随机分为纤维素生物膜组(650组)、聚砜膜组(F6组)、维生素E包被透析膜(CL-E组),每组各10例。测定透析前、后血浆丙二醛(malondialdehyde MDA)、总抗氧化能力(total anti-oxidation capacity T-AOC)、晚期蛋白氧化产物(advanced oxidation protein products AOPP)、维生素E(VitaminE VitE)水平。另选取10名终末期肾病(end stage renal disease ESRD)未透析患者及10名健康成人作为对照组(control CON)。结果与正常对照组相比,ESRD未透析组及MHD组透析前血浆IL-13、IL-18水平明显升高(P＜0.05)。血液透析后,MHD各组的血浆IL-18水平无明显变化(P＞0.05),而IL-13水平明显下降(P＜0.05)。MHD患者血浆IL-13、IL-18水平与透析龄、KT/V均无相关性(P＞0.05)。PBMC受到刺激后,ESRD未透析组培养上清中IL-13和IL-18水平及其mRNA表达均明显下降(P＜0.05);650组培养上清中IL-13水平及mRNA表达无明显变化,而IL-18显著下降(P＜0.05);F6组培养上清中IL-13及IL-18水平及其mRNA表达均显著升高(P＜0.05),但升高幅度低于健康对照组。ESRD未透析组血浆中MDA、AOPP水平较正常对照组明显升高(P＜0.05),T-AOC、VitE水平无明显变化(P＞0.05)。MHD各组透析前、后血浆AOPP水平无明显变化(P＞0.05);透析后T-AOC较透析前明显降低(P＜0.01);650及F6组透析前、后MDA水平无明显变化(P＞0.05),但650组透析前、后MDA水平明显高于ESRD未透析组(P＜0.05);CL-E组透析后MDA水平较透前明显降低(P＜0.05),VitE水平较透前明显升高(P＜0.05)。结论1、ESRD患者体内血浆IL-13、IL-18水平升高,证实ESRD患者处于微炎症状态。2、HD不能降低IL-18水平,但可降低IL-13水平。提示HD进一步减弱了患者的抗炎能力。纤维素膜与聚砜膜之间无明显差异。3、聚砜膜能改善PBMC对抗原的反应能力。故IL-13、IL-18可以作为判断透析膜的生物相容性的指标。4、ESRD患者处于氧化应激状态,维生素E包被透析膜能改善MHD患者氧化应激状态。更多还原

【Abstract】 Objective:To investigate the plasma level of interleukin-13 (IL-13)、interleukin-18 (IL-18) and the effect of different dialysis membrane on peripheral blood mononuclear cells (PBMC) synthesis and release level of IL-13、IL-18 and the oxidative stress in patients with maintanence hemodialysis (MHD). To evaluate that if IL-I3、IL-18 can be biocompatibility indexes of dialyzer.Methods:The levels of IL-I3 and IL-18 in plasma and supernatants of PBMC cultured with or without stimulation were measured by enzyme linked immunosorbent assay (ELISA) method in all groups . The expression of IL-13 and IL-18 mRNA in PBMC were evaluated by reverse transcriptase polymerase chain reaction (RT-PCR). Patients in MHD groups were dialyzed with regenerated cellulose membrane (650 group) and polysulform membrane (F6 group) respectively. At the same time, thirty patients on MHD were divided into three groups randomly. The Patients were dialyzed with regenerated cellulose membrane (650 group)、polysulform membrane (F6 group) and Vitamin E-modified membrane (CL-E group)respectively. Blood samples were collected before and after dialysis to determine the concentrations of malonylaldehyde (MDA)、vitamin E (Vit E)、total anti-oxidation capacity (T-AOC)、advanced oxidation protein products (AOPP) in plasma. Ten healthy persons and ten end stage renal disease (ESRD) patients with non-hemodialysis were as controls(CON groups).Results:The plasma levels of IL-13、IL-18 in ESRD with non-hemodialysis group and MHD group before hemodialysis were higher significantly than those of healthy controls(P<0.05). After dialysis, the plasma levels of IL-18 were not obviously changed (P>0.05) and the plasma levels of IL-13 were decreased (P<0.05) in MHD groups. The plasma lever of IL-13, IL-18 and dialysis age , KT/V have no correlation (P>0.05). After PBMC were stimulated, the level of IL-13、IL-18 significantly decreased in ESRD with non-hemodialysis group (P<0.05); in 650 group, the level of IL-13 have no obviously changes (P>0.05) and the level of IL-18 decreased(P<0.05); the level of IL-13、IL-18 in F6 group significantly increased (P<0.05), but the increased levels were lower than that in healthy controls.The plasma levels of MDA and AOPP in ESRD patients with no-dialysis were higher significantly than those of healthy controls (P<0.05). The plasma levels of T-AOC and VitE in ESRD patients with no-dialysis have no obviously changes than that in controls. The plasma levels of AOPP were no marked changes after dialysis in MHD groups (P<0.05), but obvious decline in T-AOC levels (P<0.01).There was no obviously changes in the levels of MDA in 650 group and F6 group after dialysis. The level of MDA was marked declined (P<0.05) and the level of VitE was increased (P>0.05) in CL-E group after dialysis.Conclusion:1、The plasma levels of IL-13、IL-18 in ESRD is higher than those of healthy controls. These suggest that the microinflammatory state is present in ESRD patients.2、HD can not decrease the level of IL-18, but can decrease the level of IL-13. MHD can not improve the microinflammatory state, further damage the anti-inflammatory ability, and have no significant difference between cellulose and polysulform membrane.3、The polysulform membranes can improve the immune function of PBMC. IL-13 and IL-18 can be biocompatibility indexes of dialyzer.4、The oxidative stress is present in ESRD patients. VitE - modified membrane can improve the oxidative stress in MHD.更多还原