原发性肾病综合征激素冲击治疗前后白细胞介素13及白细胞介素18基因表达
|
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。
Objective
primary 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.
Method
Thirty 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 sum
test was adopted for data processing.
Results
Comparision 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).
Conclusion
IL - 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.
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。
Objective
primary 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.
Method
Thirty 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 sum
test was adopted for data processing.
Results
Comparision 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).
Conclusion
IL - 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.
引文
1. Minty A, Chalon P, Derocq JM, Interleukin-13 is a new human lymphokine regulating inflammatory and immune responses. Nature. 1993 Mar 18; 362(6417) :248-50.
2. Zurawski G , de Vries JE. Interleukin 13, an interleukin 4-like cytokine that acts on monocytes and B cells, but not on T cells. Immunol Today, 1994;15(1) :19～26
3. Naseer T,Minshall EM,Leung DY, et al. Expression of IL-12 and IL-13 mRNA in asthma and their modulation in response to steroid therapy. Am J Respir Crit Care Med, 1997 ; 155 (3) : 845～851
4. Pouw Kraan TC, Z ee JS, Boejie LC, et al. The role of IL-13 in IgE synthesis by allegic asthma patients. [J]. Clin Exp Immunol, 1998,111:129-135
5. DeFile K M ,Jenney CR, McNally AK,et al. Interleukin -13 induces human monocyte/macrophage fusion and macrophage mannose receptor expression. J Immunol, 1997 ;158(7) :3385～3390
6. Kluth DC, Rees AJ. Inhibiting inflammatory cytokines. Semin Nephrol 1996 Nov;16(6) :576-82
7. Oksmura. H, Tsutsui H, Komatsu T et al. Cloning of a nou cytokine that induce interferon-γ. Nature, 1995,378:88
8. Ushio S, Namba M, Okura T et al. Cloning of the cDNA for human IFN -gamma- inducing factor, expression in Eschericia coli, and studies on the biologic activaties of the protein. J Immunol, 1996,156:4274-4279
9. Stoll S, Mueller G, Kurimoto M, et al. Production of IL-18 ( IFN-γ-inducing factor) messenger RNA and functional protein by murine kerationocytes [J]. Immunol, 1997,159(1) :298～302
10. Munder M, Mallo M, Eichmann K, et al. Murine microphages secrete interferon gamma upon combined stimulation with interleukin (IL)-12 and IL-18, a novel pathway of autocrine macrophage activation [J]. J Exp Med, 1998,187(12) :2103-2108
11. Pizarro TT, Michie MH, Bentz M, et al. IL-18, a novel immunoregulatory cytokine, is upregulated in Crohn disease:expression and locoalization in intestinal cells. [J]J Immunol, 1999,162(1) :6829-35
12. Gracie JA,Forsey RJ,Chan WL,et al. A proinflammatory role for IL-18 in rheumatoid arthritis. [J]J Clin Invest,1999,104(10):1393-1401
13. Romagnis S. Th1/Th2 cells. Inflamm Bowel Dis 1999 Nov;5(4) :285-94
14. HUI-KIM YAP, WANG CHEUNG et al. Th1 and Th2 Cytokine mRNA Profile in Childhood Nephrotic Syndrome: Evidence for Increased IL-13 mRNA expression in Relapse. J Am Soc Nephrol 1999, 10:529～537
15. Matsumoto K, Ohi H, Kanmatsuse K. Interleukin-10 and Interleukin-13 synergize to inhibit vascular permeability fctor release by peripheral blood mononuclear cells from patients with lipid nephrosis. Nephron, 1997,77:212～218.
16. Tomizawz S, Maruyama K, Nagasawa N, et al. Studies on vascular permeability factor derived from T-lymphocytes and inhibitory effect of plasma on its production in minimal change nephrotic syndrome. Nephron 1985,41 (2) 157-160
17. Matsumoto K. Interleukin-13 inhibits cytokin secretion by blood monocyte from patients with IgA nephropathy. Nephron, 1997,75:295～302.
18. S. P. CHEN, W. CHEUNG, C. K. HENG, S. C. JORDAN, H. K. YAP. Childhood nephrotic syndrome in relapse is associated with down-regulation of monocyte CD14 expression and lipopolysaccharide-induced tumor necrosis factor -α production. Clin Exp Immunol 2003 ; 134 : 111 - 119
19. Saura M Martinez-Dalmau R, Minty A et al. Interleukin - 13 inhibits inducible nitric oxide synthase expression in human mesangial cells. Biochem J,1996;313(2) :641～646
20. Lakkis F G, Cruet EN. Cloning of rat interleukin - 13 ( IL - 13 ) cDNA and analysis of IL- 13 gene expression in experimental glomerulonephritis Biochem Biophys Res Commun, 1993 ; 197 (2) :612～618
21. Diaz -Cazorla M, Perez-Sala D, Ros J, Regulation of cyclooxygenase -2 expression in human mesangialcells- -transcriptional inhibition by IL-13. Eur J Biochem 1999 Feb;260(1):268-74
22. Van Den Berg JG, Aten J, Claessen N, Di jkink L, Wijdenes J, Lakiss FG, Weening JJ. Interleukin- 4 and interleukin- 13 act on glomerular vixceral epithelial cells. J Am Soc Nephrol 2000 May; 11 (3) :413-22
23. Van Den Berg JG, Aten J et al. Interleukin - 14 and - 13 promote basolateral secretion of H(+) and cathepsin L by glomerular epithelial cells. Am J Physiol Renl Phsiol 2002 Jan; F26～33
24．黎磊石，刘志红。激素治疗原发性肾小球肾炎作用机制[J]。中华肾脏病杂志，1991，7：180
25. Oksmura. H, Tsutsui H, Komatsu T et al. Cloning of a nou cytokine that induce interferon-γ. Nature, 1995,378:88
26. Ushio S, Namba M, Okura T et al. Cloning of the cDNA for human IFN-gamma-inducing factor, expression in Eschericia coli, and studies on the biologic activaties of the protein. J Immunol, 1996,156:4274
27. Wong CK, Ho CY, Li EK, et al. Elevation of proinflammatory cytokine ( IL-18, IL-17, IL-12 ) and Th2 cytokine ( IL-4 ) concentrations in patients with systemic lupus [J]. Lupus, 2000,9 (8) :589-593
28. Melnikov VY, Eoder T, Fantuzzi G, et al. Impaired IL-18 processing protects caspse-1-deficient mice from ischemic acute renal failure [J]. Clin Invest,2001,107 (9) : 1145-1152
29. Melnikov VY, Faubel S, Siegmund B, et al. Neutrophil-independent mechanisms of caspase -1 - and IL-18 -mediated ischemic acute tubular necrosis in mice. J Clin Invest 2002 Oct; 110 (8) : 1083-91
30. Kitching AR,Tipping PG, Kurimoto M,et al. IL- 18 has IL- 12 -independent effects in delayed type hypersensitivity: studies in cell- mediated crescentic glomerulonephritis [J]. J Immunol,2000,165 (8) :4649-4657
31. Ozmen J, Roman D, Fountoulakis M ,et al. Eur J Immunol, 1995 ;25(1) :6～12
32. Munder M, Mallo M, Eichmann K, et al. Murine microphages secrete interferon gamma upon combined stimulation with interleukin(IL) -12 and IL-18,a novel pathway of autocrine macrophage activation [J]. J Exp Med, 1998,187 (12) :2103-2108
33. Puren A, Fantuzzi G, Gu Y, et al. Interleukin-18 ( IFN-γ-inducing fac-
tor) induces IL -1β and IL - 8 via TNF - α a production from non GD14 + human blood mononuclear cells [J]. J Clin Invest, 1998,101 (3) : 711～724
34. Takeda K, Tsutsui H, Ylshimoto T, et al. Defective NK cell activity and Th1 response in IL-18-deficient mice[J]. Immunity,1998,8(3) :383～390
35. Robison D, Shibuya K, Mui A, et al. IGIF does not drive Th1 development but synergizes with IL-12 for interfon -γ production and activates IRAK and NF-Kb[J]. Immunity, 1997,7(4) :571～581
36. Matsumoto K, Kanmatsuse K. Augmented interleukin-18 production by peripheral blood monocytes in patients with minimal change nephrotic syndrome [J]. Am J Nephron,2001,21 (1) :20-27
37. Matsumoto K, Kanmatsuse K. Elevated interleukin-18 levels in the urine of nephritic patients[J]. Nephron,2001,88(4) :334-339
38. Matsumoto K, Kanmatsuse K. interleukin-18 and interleukin-12 synergizes to stimulate the production of vascular permeability factor by T lymphocytes in normal subjects and in patients with minimal change nephrotic syndrome [J]. Nephron ,2000,85 (2): 127-133
39. HUI-KIM YAP, WANG CHEUNG et al. Th1 and Th2 Cytokine mRNA Profile in Childhood Nephrotic Syndrome: Evidence for Increased IL-13 mRNA expression in Relapse Am Soc Nephrol 1999, 10:529～537
40. Niaudef P, Habib R, Methylprednisolone pulse therapy in the treatment of severe forms of Schonlein-Henoch purpura nephritis. Pediatr Nephrol, 1998, 12:238-243
41. Makino H ,Yamasaki Y, Hayashi Y. Effect of methylprednisolone pulse therapy in patients with lupus nephritis assessed by WHO morphologic classification. Nippon Jinzo Gakkai Shi, 1990,32:791-799
42. Yorgin PD, Krasher J, Al-Uzri AY. Pulse methylprednisolone treatment of idiopathic nephrotic syndrome. Pediatr Nephrol, 2001,16:245-250
43. Mocan H, Erduran E, Karaguzel G, High dose methylprednisolone therapy in nephrotic syndrome. Indian J Pediatr, 1999,66:171-174
2. Zurawski G , de Vries JE. Interleukin 13, an interleukin 4-like cytokine that acts on monocytes and B cells, but not on T cells. Immunol Today, 1994;15(1) :19～26
3. Naseer T,Minshall EM,Leung DY, et al. Expression of IL-12 and IL-13 mRNA in asthma and their modulation in response to steroid therapy. Am J Respir Crit Care Med, 1997 ; 155 (3) : 845～851
4. Pouw Kraan TC, Z ee JS, Boejie LC, et al. The role of IL-13 in IgE synthesis by allegic asthma patients. [J]. Clin Exp Immunol, 1998,111:129-135
5. DeFile K M ,Jenney CR, McNally AK,et al. Interleukin -13 induces human monocyte/macrophage fusion and macrophage mannose receptor expression. J Immunol, 1997 ;158(7) :3385～3390
6. Kluth DC, Rees AJ. Inhibiting inflammatory cytokines. Semin Nephrol 1996 Nov;16(6) :576-82
7. Oksmura. H, Tsutsui H, Komatsu T et al. Cloning of a nou cytokine that induce interferon-γ. Nature, 1995,378:88
8. Ushio S, Namba M, Okura T et al. Cloning of the cDNA for human IFN -gamma- inducing factor, expression in Eschericia coli, and studies on the biologic activaties of the protein. J Immunol, 1996,156:4274-4279
9. Stoll S, Mueller G, Kurimoto M, et al. Production of IL-18 ( IFN-γ-inducing factor) messenger RNA and functional protein by murine kerationocytes [J]. Immunol, 1997,159(1) :298～302
10. Munder M, Mallo M, Eichmann K, et al. Murine microphages secrete interferon gamma upon combined stimulation with interleukin (IL)-12 and IL-18, a novel pathway of autocrine macrophage activation [J]. J Exp Med, 1998,187(12) :2103-2108
11. Pizarro TT, Michie MH, Bentz M, et al. IL-18, a novel immunoregulatory cytokine, is upregulated in Crohn disease:expression and locoalization in intestinal cells. [J]J Immunol, 1999,162(1) :6829-35
12. Gracie JA,Forsey RJ,Chan WL,et al. A proinflammatory role for IL-18 in rheumatoid arthritis. [J]J Clin Invest,1999,104(10):1393-1401
13. Romagnis S. Th1/Th2 cells. Inflamm Bowel Dis 1999 Nov;5(4) :285-94
14. HUI-KIM YAP, WANG CHEUNG et al. Th1 and Th2 Cytokine mRNA Profile in Childhood Nephrotic Syndrome: Evidence for Increased IL-13 mRNA expression in Relapse. J Am Soc Nephrol 1999, 10:529～537
15. Matsumoto K, Ohi H, Kanmatsuse K. Interleukin-10 and Interleukin-13 synergize to inhibit vascular permeability fctor release by peripheral blood mononuclear cells from patients with lipid nephrosis. Nephron, 1997,77:212～218.
16. Tomizawz S, Maruyama K, Nagasawa N, et al. Studies on vascular permeability factor derived from T-lymphocytes and inhibitory effect of plasma on its production in minimal change nephrotic syndrome. Nephron 1985,41 (2) 157-160
17. Matsumoto K. Interleukin-13 inhibits cytokin secretion by blood monocyte from patients with IgA nephropathy. Nephron, 1997,75:295～302.
18. S. P. CHEN, W. CHEUNG, C. K. HENG, S. C. JORDAN, H. K. YAP. Childhood nephrotic syndrome in relapse is associated with down-regulation of monocyte CD14 expression and lipopolysaccharide-induced tumor necrosis factor -α production. Clin Exp Immunol 2003 ; 134 : 111 - 119
19. Saura M Martinez-Dalmau R, Minty A et al. Interleukin - 13 inhibits inducible nitric oxide synthase expression in human mesangial cells. Biochem J,1996;313(2) :641～646
20. Lakkis F G, Cruet EN. Cloning of rat interleukin - 13 ( IL - 13 ) cDNA and analysis of IL- 13 gene expression in experimental glomerulonephritis Biochem Biophys Res Commun, 1993 ; 197 (2) :612～618
21. Diaz -Cazorla M, Perez-Sala D, Ros J, Regulation of cyclooxygenase -2 expression in human mesangialcells- -transcriptional inhibition by IL-13. Eur J Biochem 1999 Feb;260(1):268-74
22. Van Den Berg JG, Aten J, Claessen N, Di jkink L, Wijdenes J, Lakiss FG, Weening JJ. Interleukin- 4 and interleukin- 13 act on glomerular vixceral epithelial cells. J Am Soc Nephrol 2000 May; 11 (3) :413-22
23. Van Den Berg JG, Aten J et al. Interleukin - 14 and - 13 promote basolateral secretion of H(+) and cathepsin L by glomerular epithelial cells. Am J Physiol Renl Phsiol 2002 Jan; F26～33
24．黎磊石，刘志红。激素治疗原发性肾小球肾炎作用机制[J]。中华肾脏病杂志，1991，7：180
25. Oksmura. H, Tsutsui H, Komatsu T et al. Cloning of a nou cytokine that induce interferon-γ. Nature, 1995,378:88
26. Ushio S, Namba M, Okura T et al. Cloning of the cDNA for human IFN-gamma-inducing factor, expression in Eschericia coli, and studies on the biologic activaties of the protein. J Immunol, 1996,156:4274
27. Wong CK, Ho CY, Li EK, et al. Elevation of proinflammatory cytokine ( IL-18, IL-17, IL-12 ) and Th2 cytokine ( IL-4 ) concentrations in patients with systemic lupus [J]. Lupus, 2000,9 (8) :589-593
28. Melnikov VY, Eoder T, Fantuzzi G, et al. Impaired IL-18 processing protects caspse-1-deficient mice from ischemic acute renal failure [J]. Clin Invest,2001,107 (9) : 1145-1152
29. Melnikov VY, Faubel S, Siegmund B, et al. Neutrophil-independent mechanisms of caspase -1 - and IL-18 -mediated ischemic acute tubular necrosis in mice. J Clin Invest 2002 Oct; 110 (8) : 1083-91
30. Kitching AR,Tipping PG, Kurimoto M,et al. IL- 18 has IL- 12 -independent effects in delayed type hypersensitivity: studies in cell- mediated crescentic glomerulonephritis [J]. J Immunol,2000,165 (8) :4649-4657
31. Ozmen J, Roman D, Fountoulakis M ,et al. Eur J Immunol, 1995 ;25(1) :6～12
32. Munder M, Mallo M, Eichmann K, et al. Murine microphages secrete interferon gamma upon combined stimulation with interleukin(IL) -12 and IL-18,a novel pathway of autocrine macrophage activation [J]. J Exp Med, 1998,187 (12) :2103-2108
33. Puren A, Fantuzzi G, Gu Y, et al. Interleukin-18 ( IFN-γ-inducing fac-
tor) induces IL -1β and IL - 8 via TNF - α a production from non GD14 + human blood mononuclear cells [J]. J Clin Invest, 1998,101 (3) : 711～724
34. Takeda K, Tsutsui H, Ylshimoto T, et al. Defective NK cell activity and Th1 response in IL-18-deficient mice[J]. Immunity,1998,8(3) :383～390
35. Robison D, Shibuya K, Mui A, et al. IGIF does not drive Th1 development but synergizes with IL-12 for interfon -γ production and activates IRAK and NF-Kb[J]. Immunity, 1997,7(4) :571～581
36. Matsumoto K, Kanmatsuse K. Augmented interleukin-18 production by peripheral blood monocytes in patients with minimal change nephrotic syndrome [J]. Am J Nephron,2001,21 (1) :20-27
37. Matsumoto K, Kanmatsuse K. Elevated interleukin-18 levels in the urine of nephritic patients[J]. Nephron,2001,88(4) :334-339
38. Matsumoto K, Kanmatsuse K. interleukin-18 and interleukin-12 synergizes to stimulate the production of vascular permeability factor by T lymphocytes in normal subjects and in patients with minimal change nephrotic syndrome [J]. Nephron ,2000,85 (2): 127-133
39. HUI-KIM YAP, WANG CHEUNG et al. Th1 and Th2 Cytokine mRNA Profile in Childhood Nephrotic Syndrome: Evidence for Increased IL-13 mRNA expression in Relapse Am Soc Nephrol 1999, 10:529～537
40. Niaudef P, Habib R, Methylprednisolone pulse therapy in the treatment of severe forms of Schonlein-Henoch purpura nephritis. Pediatr Nephrol, 1998, 12:238-243
41. Makino H ,Yamasaki Y, Hayashi Y. Effect of methylprednisolone pulse therapy in patients with lupus nephritis assessed by WHO morphologic classification. Nippon Jinzo Gakkai Shi, 1990,32:791-799
42. Yorgin PD, Krasher J, Al-Uzri AY. Pulse methylprednisolone treatment of idiopathic nephrotic syndrome. Pediatr Nephrol, 2001,16:245-250
43. Mocan H, Erduran E, Karaguzel G, High dose methylprednisolone therapy in nephrotic syndrome. Indian J Pediatr, 1999,66:171-174