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| 骨桥蛋白和核转录因子在慢性环孢素A肾毒性中的作用 |
| 方梅荣,金英顺,金吉哲,崔镇花,金海峰,郑海兰,李锦姬,姜玉姬,金华,李灿* |
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(延边大学附属医院肾脏内科, 延吉 133000
*通信作者) |
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| 摘要: |
| 目的 探讨炎性介质骨桥蛋白（OPN）的表达和核转录因子在慢性环孢素A（CsA）肾毒性中的作用。方法 雄性Sprague-Dawley大鼠喂食低盐 （0.05% 钠盐）饲料下分为两组，正常对照组给予皮下注射橄榄油（1 mL·kg-1·d-1）；毒性组给予皮下注射CsA （15 mg·kg-1·d-1） 4周。通过观察炎性细胞浸润（ED-1） 和肾小管间质纤维化，比较两组大鼠的肾小管间质损伤程度；采用RNA印迹、免疫组织化学染色方法检测OPN在mRNA和蛋白水平的表达；用凝胶电泳迁移率实验 （EMSA）分析法和免疫印迹法测定核转录因子（NF-κB 和 AP-1）的结合活性和IκB蛋白的表达。结果 毒性组大鼠表现为肾小管间质带状纤维化[（38.9±3.3）%/5 mm2vs（0±0）%/5 mm2，P<0.01]和大量ED-1阳性细胞浸润[（89±9）vs（7±2），P<0.01]。与对照组相比，毒性组大鼠 OPN mRNA和蛋白的表达增加[（729±37）%vs（103±4）%，P<0.01]，分布于肾小管上皮细胞，其主要位于肾小管间质损伤部位；毒性组核转录因子NF-κB [（218±19）%vs（116±15）%，P<0.01]和AP-1 [（735±225）%vs（101±4）%，P<0.01]结合活性增加，而IκB蛋白的表达减少[（9±7）%vs（105±7）%，P<0.01]。直线相关分析示OPN mRNA的表达与肾小管间质纤维化程度（r=0.959，P<0.001）和核转录因子的结合活性呈正相相关（NF-κB： r=0.773，P<0.01；AP-1： r=0.619，P=0.01）。结论 炎性介质OPN和核转录因子参与了慢性CsA肾毒性肾小管间质的损伤。 |
| 关键词: 慢性环孢素A肾毒性 骨桥蛋白 NF-κB 转录因子AP-1 |
| DOI：10.3724/SP.J.1008.2014.00476 |
| 投稿时间：2013-11-04修订日期：2014-01-13 |
| 基金项目:国家自然科学基金（81160092）. |
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| Roles of osteopontin and nuclear transcription factor in chronic cyclosporine nephrotoxicity |
| FANG Mei-rong,JIN Ying-shun,JIN Ji-zhe,CUI Zhen-hua,JIN Hai-feng,ZHENG Hai-lan,LI Jin-ji,JIANG Yu-ji,JIN Hua,LI Can* |
(Department of Nephrology, Affiliated Hospital of Yanbian University, Yanji 133000, Jilin, China
*Corresponding authors.) |
| Abstract: |
| Objective To investigate the roles of osteopontin(OPN) and nuclear transcription factor in a rat model of chronic cyclosporine A (CsA) nephrotoxicity. Methods Male Sprague-Dawley rats maintained on a low salt diet were treated daily with vehicle (olive oil, 1 mL·kg-1·d-1, s.c.) and CsA (15 mL·kg-1·d-1, s.c.) for 4 weeks. Renal histopathology was estimated by trichrome staining (tubulointerstitial fibrosis) and immunohistochemistry (ED-1) to assess the degrees of renal tubulointerstitial lesions. In addition, OPN mRNA and protein expression and nuclear transcription factor (NF-κB and AP-1) were studied by northern blotting analysis, immunohistochemistry, electrophoretic mobility shift assay, and immunoblotting analysis. Results CsA-treated rats displayed significantly striped tubulointerstitial fibrosis ([38.9±3.3]%/5 mm2 vs [0±0]%/5 mm2, P<0.01) and increased ED-1-positive cells (89±9 vs 7±2, P<0.01). Compared with VH-treated rats, CsA-treated rats showed significantly upregulated OPN mRNA and protein expression in the proximal tubular cells, mainly localizing at areas of severe injured tissues. CsA-treated group also had significantly increased activities of NF-κB ([218±19]% vs [116±15]%, P<0.01) and AP-1 ([735±225]% vs [101±4]%, P<0.01), and significantly decreased expression of ([9±7]% vs [105±7]%, P<0.01). Correlation analysis revealed that upregulated OPN mRNA was positively correlated with tubulointerstitial fibrosis (r= 0.959, P<0.001) and activities of NF-κB and AP-1 (NF-κB: r=0.773, P<0.01; AP-1: r=0.619, P=0.01, respectively). Conclusion Our findings suggest that OPN, nuclear transcription factor NF-κB and AP-1 are involved in renal tubulointerstitial injury in chronic CsA nephrotoxicity. |
| Key words: chronic cyclosporine A nephrotoxicity osteopontin NF-κB transcription factor AP-1 |
