骨桥蛋白和核转录因子在慢性环孢素A肾毒性大鼠肾组织中的作用

发布时间：2018-09-03 14:37

【摘要】：目的：探讨炎性介质骨桥蛋白(Osteopontin,oPN)的表达和核转录因子在大鼠慢性环孢素A(CsA)肾毒性中的作用。方法：雄性Sprague-Dawley大鼠喂食低盐饲料(0.05%钠盐)下分为两组,正常对照组给予皮下注射橄榄油(1m1/kg/d)；慢性CsA肾毒性组给予皮下注射环孢素A(15mg/kg/d)4周。通过三色染色观察肾小管间质纤维化程度和免疫组化观察ED-1阳性细胞；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(Elecrophoretic mobility shift assay, 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：与对照组相比,毒性组大鼠表现为肾小管间质带状纤维化[(38.9±3.3)%vs.(0.0±0.0)%,P0.01]和大量ED-1阳性细胞浸润[(89±9)%vs.(7±2)%,P0.01]。与对照组相比,毒性组大鼠OPN mRNA和蛋白的表达明显增加[(729±37)%vs.(103±4)%,P0.01],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±19vs.116±15,P0.01)和AP-1(735±225vs.101±4,P0.01)结合活性显著增加,而I KB蛋白的表达减少(9±7vs.105±7,P0.01)。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,P0.001)和核转录因子的结合活性呈正相相关(NF-κB:r=0.773,尸0.01:AP.1:r=:0.619,P=0.01).结论：在慢性环孢素A肾毒性中OPN表达的上调与肾小管间质纤维化程度相关,且可能与NF-κB和AP-1的激活密切相关。
[Abstract]:AIM: To investigate the expression of osteopontin (oPN) and the role of nuclear transcription factors in chronic cyclosporine A (CsA) nephrotoxicity in rats.
Methods: Male Sprague-Dawley rats were fed low-salt diet (0.05% sodium salt) and divided into two groups. Normal control group was given subcutaneous injection of olive oil (1m1/kg/d); chronic CsA nephrotoxicity group was given subcutaneous injection of cyclosporine A (15mg/kg/d) for 4 weeks. The expression of OPN at mRNA and protein levels was detected by RNA imprinting and immunohistochemical staining, and the binding activity of nuclear transcription factors (NF-kappa B and AP-1) and the expression of I-kappa B protein were determined by gel electrophoretic mobility shift assay (EMSA) and immunoblotting.
Results: Compared with the control group, the toxic group showed renal tubulointerstitial banded fibrosis [(38.9 +3.3)% vs. (0.0 +0.0)%, P 0.01] and a large number of ED-1 positive cell infiltration [(89 +9)% vs. (7 +2)%, P 0.01]. Compared with the control group, the expression of OPN mRNA and protein in the toxic group increased significantly [(729 +37)% vs. (103 +4)%, P 0.01], and distributed in the renal tubules. In the toxic group, the binding activity of nuclear transcription factor NF-kappa B (218 65507 CONCLUSION: The up-regulation of OPN expression in chronic cyclosporine A nephrotoxicity is associated with the degree of tubulointerstitial fibrosis and may be closely related to the activation of NF-kappa B and AP-1.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692

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