【作者】 姜红堃；

【导师】 姜红；

【作者基本信息】 中国医科大学 ， 儿科学， 2004， 硕士

【摘要】 目的 原发性肾病综合征(PNS)是小儿常见的肾脏疾病，其病程中常有反复或复发，发病机理仍不十分清楚，研究结果的多样性提示其发病机理的异质性。本实验通过观察白细胞介素13(IL-13)及白细胞介素18(IL-18)在小儿PNS中的变化探讨其在PNS发病中的变化及甲基强的松龙冲击治疗(MPT)对IL-13及IL-18表达的影响。 方法 选择自2002年10月至2003年12月期间在我院儿科住院PNS患者30例为观察组，且之前均未用过或至少3个月内未用糖皮质激素及免疫抑制剂治疗。其中男18例，女12例，年龄2～14岁，平均7.5岁。选择门诊健康体检儿童20例为正常对照组，男13例，女7例，年龄3～15岁，平均8岁。所有患者均予甲基强的松龙15-30mg/Kg静脉滴注，连续三天，而后长期口服强的松。采用酶联免疫吸附(ELISA)法测定20例正常儿童及30例PNS患儿MPT前、结束后(第二天、第五天)血清IL-13及IL-18水平，操作参照使用说明。人IL-13及IL-18试剂盒购于深圳晶美生物工程有限公司，根据标准曲线算出各血清标本的IL-13及IL-18浓度。采用RT-PCR法检测20例正常儿及20例PNS患儿MPT前、结束后(第二天、第五天)外周血单个核细胞(PBMC)IL-13及IL-18mRNA水平。反转录试剂盒购于大连TAKARA公司，Trizol购于GIBCO BRL USA公司。同时采用双缩脲法检测24小时尿蛋白量。利用SPSS软件采用秩和检验统计方法进行分析处理。 结果 (1)治疗前PNS患儿血清中IL-13蛋白水平(中位数38.48pg/ml)及IL-13mRNA表达(中位数1.31)明显高于MPT结束后第五天和正常对照组(P均<0.01);而MPr结束后第五天血清中IL一13蛋白水平(中位数巧.33p扩耐)及IL一13mRNA表达(中位数0.89)与正常对照组比较差异有显著性(P<0.05);(2)治疗前PNS患儿血清中IL一18蛋白水平(中位数149.22p扩血)及IL一18mRNA表达(中位数1.25)明显高于MPr结束后第五天组和正常对照组(P均<0.01);而MPr结束后第五天组血清中IL一18蛋白水平(中位数80.01p扩血)及IL一 18mRNA表达(中位数0.67)与正常对照组比较差异有显著性;(3) MPr前及结束后第二天IL一13及IL一18血清蛋白水平及mRNA表达差异无显著性(P均>0.05);(4) MPr结束后第二天及第五天IL一13及IL一18血清蛋白水平及mRNA表达差异有显著性(P均<0.01);(5)PNS患儿血清中IL一13及IL一18水平与24小时尿蛋白量呈正相关。结论 IL一13及IL一18在PNS的发病机制中发挥着重要作用，M盯对肾病综合征患儿IL一13、IL一18在蛋白合成和基因转录两个水平上有抑制作用，测定血清中IL一13及IL一18水平及PBMCIL一13mRNA、IL一18mRNA表达可作为观察病情活动状态的一个免疫学指标，具有一定的临床意义。更多还原

【Abstract】 Objectiveprimary nephrotic syndrome ( PNS) is one of the common renal disease in children, the pathogenesis of which is unclear. The purpose of the study was to evaluate the serum concentration and mRNA expression of IL - 13 and IL - 18 before and after the methylprednisolone pulse therapy ( MPT) in PNS.MethodThirty children with PNS diagnosed from October 2002 to December 2003 were enrolled in this study ( patients group). They were not treated with glu-cocorticoid and other immunosurppressives at least within the recent 3 months. The children aged from 2 to 14 years (mean 7. 5 years) and included 18 boys and 12 girls. 20 healthy children were selected as control group after physical examination. The children in control group aged from 3 to 15 years old ( mean 8years) and included 13 boys and 7 girls. All patients were treated with MPT intravenenously (15 ~30mg/Kg) for successive 3 days followed by oral predni-sone. The serum protein level of IL -13 and IL - 18 was measured by ELISA according to the manufacturers instructions. Human IL - 13 and IL - 18 ELISA kits were purchased from Jingmei Corporation Shenzhen, China. And the concentration was obtained after drawing the standard curve. The expression of IL - 13 and IL - 18 gene was detected with RT - PCR method. The important reverse transcription reagent kits were bought from TAKARA corporation Dalian and Tr-izol reagent was bought from GIBCO BRL, USA. Statistical analysis of rank sumtest was adopted for data processing.ResultsComparision of the serum IL -13 and IL - 18 levels in the same patient before and after the therapy showed significant difference( both P <0.01). The serum level in patient group before the therapy increased obviously in comparison to the level of the control group. The serum level of IL - 13 and IL - 18 in patient group after the therapy also showed significant difference compared to the control group(P <0. 01) . The mRNA expression of IL - 13 and IL - 18 in the same patient before and after therapy showed significant difference (both P <0. 01).ConclusionIL - 13 and IL - 18 maybe involved in the pathogenesis of PNS because of the significant increase of the serum IL - 13 and IL - 18 level and PBMC IL - 13 and IL - 18 mRNA expression in PNS children. Methylprednisolone pulse therapy in PNS was able to inhibit the protein production and PBMC mRNA expression of IL - 13 and IL - 18 ,so the therapeutic mechanism of MPT in PNS might be associated with the inhibition of IL - 13 and IL - 18 expression.更多还原